Pain is a subjective and unpleasant sensation that significantly impacts the daily lives of individuals.Chronic pain represents one of the most challenging public health issues,and ensuring effective pain management i...Pain is a subjective and unpleasant sensation that significantly impacts the daily lives of individuals.Chronic pain represents one of the most challenging public health issues,and ensuring effective pain management is a fundamental right of individuals and a sacred duty of healthcare providers.Cannabis,one of the earliest recognized medicinal plants,contains cannabinoids,which are non-opioid substances that modulate nociceptive responses.Electroacupuncture(EA),characterized by its low-risk and well-tolerated nature,is pivotal in pain management.The endocannabinoid system consists of endocannabinoids,cannabinoid receptors,and enzymes involved in endocannabinoid synthesis,degradation,and transport.Recently,the role of the endocannabinoid system in pain development and EA analgesia has attracted considerable research attention.Studies have highlighted the role of the endocannabinoid system in various types of pain,including inflammatory pain,neuropathic pain,and cancer-related pain,as well as in EA analgesia.This study aims to review the mechanisms of endocannabinoid system involvement in pain modulation and EA analgesia to provide insights to inform clinical approaches to pain management.展开更多
Background As cows transition from pregnancy to lactation,free fatty acids(FFA)are mobilized from adipose tissues(AT)through lipolysis to counter energy deficits.In clinically healthy cows,lipolysis intensity is reduc...Background As cows transition from pregnancy to lactation,free fatty acids(FFA)are mobilized from adipose tissues(AT)through lipolysis to counter energy deficits.In clinically healthy cows,lipolysis intensity is reduced throughout lactation;however,if F FA release exceeds tissue demands or the liver's metabolic capacity,lipid byproducts accumulate,incre a sing cows'risk of metabolic and infectious disease.Endocannabinoids(eCBs)and their congeners,N-acylethanolamines(NAEs),are lipid-based compounds that modulate metabolism and inflammation.Their synthesis and release depend upon the availability of FFA precursors and the abundance of synthesizing and degrading enzymes and transporters.Therefore,we hypothesized that eCB production and transcription of endocannabinoid system components are modulated by lipolysis pathways in adipocytes.To test this hypothesise we stimulated canonical(isoproterenol,1μmol/L;ISO)and inflammatory(lipopolysaccharide,1μg/m L;LPS)lipolysis pathways in adipocytes isolated from the AT of 5 Holstein dairy cows.Following,we assessed lipolysis intensity,adipocytes'release of eCBs,and transcription of endocannabinoid system components.Results We found that ISO and LPS stimulated lipolysis at comparable intensities.Exposure to either treatment tended to elevate the release of eCBs and NAEs by cultured adipocytes;however,specific eCBs and NAEs and the transcriptional profiles differed by treatment.On one hand,ISO enhanced adipocytes'release of 2-arachidonoylglycerol(2-AG)but reduced NAE production.Klotably,ISO enhanced the cells"expression of enzymes associated with 2-AG biosynthesis(INPP5F,GDPD5,GPAT4),transport(CD36),and adipogenesis(PPARG).Conversely,LPS enhanced adipocytes'synthesis and release of N-arachidonoylethanolamide(AEA).This change coincided with enhanced transcription of the NAE-biosynthesizing enzyme,PTPN22,and adipocytes'transcription of genes related to eCB degradation(PTGS2,MGLL,CYP27B1).Furthermore,LPS enhanced adipocytes'transcription of eCB and NAE transporters(HSPA1A,SCP2)and the expression of the anti-adipogenic ion channel,TRPV3.Conclusions Our data provide evidence for distinct modulatory roles of canonical and inflammatory lipolysis pathways over eCB release and transcriptional regulation of biosynthesis,degradation,transport,and ECS signaling in cows'adipocytes.Based on our findings,we conclude that,within adipocytes,eCB production and ECS component expression are,at least in part,mediated by lipolysis in a pathway-dependent manner.These findings contribute to a deeper understanding of the molecular mechanisms underlying metabolic regulation in dairy cows'AT,with potential implications for prevention and treatment of inflammatory and metabolic disorders.展开更多
Background The placenta plays a crucial role in supporting and influencing fetal development.We compared the effects of prepartum supplementation with omega-3(n-3)fatty acid(FA)sources,flaxseed oil(FLX)and fish oil(FO...Background The placenta plays a crucial role in supporting and influencing fetal development.We compared the effects of prepartum supplementation with omega-3(n-3)fatty acid(FA)sources,flaxseed oil(FLX)and fish oil(FO),on the expression of genes and proteins related to lipid metabolism,inflammation,oxidative stress,and the endocannabinoid system(ECS)in the expelled placenta,as well as on FA profile and inflammatory response of neonates.Late-pregnant Holstein dairy cows were supplemented with saturated fat(CTL),FLX,or FO.Placental cotyledons(n=5)were collected immediately after expulsion,and extracted RNA and proteins were analyzed by RTPCR and proteomic analysis.Neonatal blood was assessed for FA composition and concentrations of inflammatory markers.Results FO increased the gene expression of fatty acid binding protein 4(FABP4),interleukin 10(IL-10),catalase(CAT),cannabinoid receptor 1(CNR1),and cannabinoid receptor 2(CNR2)compared with CTL placenta.Gene expression of ECS-enzyme FA-amide hydrolase(FAAH)was lower in FLX and FO than in CTL.Proteomic analysis identified 3,974 proteins;of these,51–59 were differentially abundant between treatments(P≤0.05,|fold change|≥1.5).Top canonical pathways enriched in FLX vs.CTL and in FO vs.CTL were triglyceride metabolism and inflammatory processes.Both n-3 FA increased the placental abundance of FA binding proteins(FABPs)3 and 7.The abundance of CNR1 cannabinoid-receptor-interacting-protein-1(CNRIP1)was reduced in FO vs.FLX.In silico modeling affirmed that bovine FABPs bind to endocannabinoids.The FLX increased the abundance of inflammatory CD44-antigen and secreted-phosphoprotein-1,whereas prostaglandin-endoperoxide synthase 2 was decreased in FO vs.CTL placenta.Maternal FO enriched neonatal plasma with n-3 FAs,and both FLX and FO reduced interleukin-6 concentrations compared with CTL.Conclusion Maternal n-3 FA from FLX and FO differentially affected the bovine placenta;both enhanced lipid metabolism and modulated oxidative stress,however,FO increased some transcriptional ECS components,possibly related to the increased FABPs.Maternal FO induced a unique balance of pro-and anti-inflammatory components in the placenta.Taken together,different sources of n-3 FA during late pregnancy enhanced placental immune and metabolic processes,which may affect the neonatal immune system.展开更多
Cannabinoids are lipophilic substances derived from Cannabis sativa that can exert a variety of effects in the human body.They have been studied in cellular and animal models as well as in human clinical trials for th...Cannabinoids are lipophilic substances derived from Cannabis sativa that can exert a variety of effects in the human body.They have been studied in cellular and animal models as well as in human clinical trials for their therapeutic benefits in several human diseases.Some of these include central nervous system(CNS)diseases and dysfunctions such as forms of epilepsy,multiple sclerosis,Parkinson’s disease,pain and neuropsychiatric disorders.In addition,the endogenously produced cannabinoid lipids,endocannabinoids,are critical for normal CNS function,and if controlled or modified,may represent an additional therapeutic avenue for CNS diseases.This review discusses in vitro cellular,ex vivo tissue and in vivo animal model studies on cannabinoids and their utility as therapeutics in multiple CNS pathologies.In addition,the review provides an overview on the use of cannabinoids in human clinical trials for a variety of CNS diseases.Cannabinoids and endocannabinoids hold promise for use as disease modifiers and therapeutic agents for the prevention or treatment of neurodegenerative diseases and neurological disorders.展开更多
To sustain the nutrient demands of rapid fetal growth,parturition,and milk synthesis,periparturient dairy cows mobilize adipose tissue fatty acid stores through lipolysis.This process induces an inflammatory response ...To sustain the nutrient demands of rapid fetal growth,parturition,and milk synthesis,periparturient dairy cows mobilize adipose tissue fatty acid stores through lipolysis.This process induces an inflammatory response within AT that is resolved as lactation progresses;however,excessive and protracted lipolysis compounds the risk for metabolic and inflammatory diseases.The suppression of lipolytic action and inflammation,along with amplification of adipogenesis and lipogenesis,serve as prospective therapeutic targets for improving the health of periparturient dairy cows.Generally,the activation of cannabinoid receptors by endocannabinoids enhances adipogenesis and lipogenesis,suppresses lipolysis,and increases appetite in mammals.These biological effects of activating the endocannabinoid system open the possibility of harnessing the endocannabinoid system through nutritional intervention in dairy herds as a potential tool to improve dairy cows’health,although much is still to be revealed in this context.This review summarizes the current knowledge surrounding the components of the endocannabinoid system,elaborates on the metabolic effects of its activation,and explores the potential to modulate its activity in periparturient dairy cows.展开更多
Being a great threaten for human health, obesity has become a pandemic chronic disease. There have been several therapeutic treatments for this social health issue, including diet and exercise therapy, medication and ...Being a great threaten for human health, obesity has become a pandemic chronic disease. There have been several therapeutic treatments for this social health issue, including diet and exercise therapy, medication and surgery, among which the diet is still the most common way. However, none of these therapeutic measures available is ideal, making it necessary to find an effective medical treatment. The endocannabinoid system, which is well known for its contributions in certain mental processes such as relaxation, amelioration of pain and anxiety, and sedation initiation, has been recently reported to play an essential role in regulating appetite and metabolism to maintain energy balance, leading to the belief that endocannabinoid system is closely related to obesity. This new discovery deepens our understanding of obesity, and provides us with a new direction for clinical obesity treatment. Rimonabant is an antagonist for CB1, and has entered the market in some countries. However, although effective as an anti-obesity drug, rimonabant also causes obviously adverse side-effects, thus is being doubted and denied for medical usage.展开更多
The endocannabinoid system(ECS),particularly its signaling pathways and ligands,has garnered considerable interest in recent years.Along with clinical work investigating the ECS’functions,including its role in the de...The endocannabinoid system(ECS),particularly its signaling pathways and ligands,has garnered considerable interest in recent years.Along with clinical work investigating the ECS’functions,including its role in the development of neurological and inflammatory conditions,much research has focused on developing analytical protocols enabling the precise monitoring of the levels and metabolism of the most potent ECS ligands:exogenous phytocannabinoids(PCs)and endogenous cannabinoids(endocannabinoids,ECs).Solid-phase microextraction(SPME)is an advanced,non-exhaustive sample-preparation technique that facilitates the precise and efficient isolation of trace amounts of analytes,thus making it appealing for the analysis of PCs and ECs in complex matrices of plant and animal/human origin.In this paper,we review recent forensic medicine and toxicological studies wherein SPME has been applied to monitor levels of PCs and ECs in complex matrices,determine their effects on organism physiology,and assess their role in the development of several diseases.展开更多
Diabetic complications,chiefly seen in long-term situations,are persistently deleterious to a large extent,requiring multi-factorial risk reduction strategies beyond glycemic control.Diabetic cardiomyopathy is one of ...Diabetic complications,chiefly seen in long-term situations,are persistently deleterious to a large extent,requiring multi-factorial risk reduction strategies beyond glycemic control.Diabetic cardiomyopathy is one of the most common deleterious diabetic complications,being the leading cause of mortality among diabetic patients.The mechanisms of diabetic cardiomyopathy are multi-factorial,involving increased oxidative stress,accumulation of advanced glycation end products(AGEs),activation of various pro-inflammatory and cell death signaling pathways,and changes in the composition of extracellular matrix with enhanced cardiac fibrosis.The novel lipid signaling system,the endocannabinoid system,has been implicated in the pathogenesis of diabetes and its complications through its two main receptors:Cannabinoid receptor type 1 and cannabinoid receptor type 2,alongside other components.However,the role of the endocannabinoid system in diabetic cardiomyopathy has not been fully investigated.This review aims to elucidate the possible mechanisms through which cannabinoids and the endocannabinoid system could interact with the pathogenesis and the development of diabetic cardiomyopathy.These mechanisms include oxidative/nitrative stress,inflammation,accumulation of AGEs,cardiac remodeling,and autophagy.A better understanding of the role of cannabinoids and the endocannabinoid system in diabetic cardiomyopathy may provide novel strategies to manipulate such a serious diabetic complication.展开更多
Background:Dietary supplementation of omega-3 fatty acids can reduce the activation of the endocannabinoid system(ECS)by decreasing the availability of arachidonic acid,thus lowering endocannabinoids(e CBs)levels.The ...Background:Dietary supplementation of omega-3 fatty acids can reduce the activation of the endocannabinoid system(ECS)by decreasing the availability of arachidonic acid,thus lowering endocannabinoids(e CBs)levels.The ECS is a modulator of energy metabolism,stress response and inflammation in mammals,yet there is little information on the roles of the ECS in transition dairy cows.During the periparturient period,the adipose tissue and liver are the main metabolic organs that participate in the adaptations of dairy cows to onset of lactation;however,exceeded adipose tissue lipolysis and accumulation of lipids in the liver have adverse effects on cows'physiology.Here we aimed to examine whether omega-3 supplementation during the transition period will modulate ECS activation and affect metabolic and inflammatory indices in postpartum dairy cows,by supplementing twenty-eight transition Holstein dairy cows with either saturated fat(CTL)or encapsulated flaxseed oil(FLX).Components of the ECS,metabolic and inflammatory markers were measured in blood,liver,and subcutaneous adipose tissue.Results:FLX supplementation reduced feed intake by 8.1%(P<0.01)and reduced plasma levels of arachidonic acid(by 44.2%;P=0.02)and anandamide(by 49.7%;P=0.03)postpartum compared to CTL.The m RNA transcription levels of the cannabinoid receptor 1(CNR1/CB1)tended to be lower(2.5 folds)in white blood cells of FLX than in CTL(P=0.10),and protein abundance of ECS enzyme monoacylglycerol lipase was higher in peripheral blood mononuclear cells of FLX than in CTL(P=0.04).In adipose tissue,palmitoylethanolamide levels were lower in FLX than in CTL(by 61.5%;P=0.02),relative m RNA transcription of lipogenic genes were higher,and the protein abundance of cannabinoid receptor 2(P=0.08)and monoacylglycerol lipase(P=0.10)tended to be higher in FLX compared to CTL.Hepatic 2-arachidonoylglycerol tended to be higher(by 73.1%;P=0.07),and interlukin-6 m RNA transcription level was 1.5 folds lower in liver of FLX than in CTL(P=0.03).Conclusions:Nutritional supplementation of omega-3 fatty acids seems to partly modulate ECS activation,which could be related to lower feed intake.The altered ECS components in blood,adipose tissue and liver are associated with moderate modulations in lipid metabolism in the adipose and inflammation in liver of peripartum dairy cows.展开更多
Humans have been using Cannabis and its extracts for a few thousand years as a medicinal and recreational drug. How- ever, the chemical component in Cannabis sativa, △9-tet- rahydrocannabinol （△9-THC）, an exogenou...Humans have been using Cannabis and its extracts for a few thousand years as a medicinal and recreational drug. How- ever, the chemical component in Cannabis sativa, △9-tet- rahydrocannabinol （△9-THC）, an exogenous cannabinoid, remained unknown until it was isolated and identified as the main psychoactive ingredient （Gaoni and Mechoulam, 1964）.展开更多
Globally,it is evident that glioblastoma multiforme(GBM)is an aggressive malignant cancer with a high mortality rate and no effective treatment options.Glioblastoma is classified as the stage-four progression of a gli...Globally,it is evident that glioblastoma multiforme(GBM)is an aggressive malignant cancer with a high mortality rate and no effective treatment options.Glioblastoma is classified as the stage-four progression of a glioma tumor,and its diagnosis results in a shortened life expectancy.Treatment options for GBM include chemotherapy,immunotherapy,surgical intervention,and conventional pharmacotherapy;however,at best,they extend the patient’s life by a maximum of 5 years.GBMs are considered incurable due to their high recurrence rate,despite various aggressive therapeutic approaches which can have many serious adverse effects.Ceramides,classified as endocannabinoids,offer a promising novel therapeutic approach for GBM.Endocannabinoids may enhance the apoptosis of GBM cells but have no effect on normal healthy neural cells.Cannabinoids promote atypical protein kinase C,deactivate fatty acid amide hydrolase enzymes,and activate transient receptor potential vanilloid 1(TRPV1)and TRPV2 to induce pro-apoptotic signaling pathways without increasing endogenous cannabinoids.In previous in vivo studies,endocannabinoids,chemically classified as amide formations of oleic and palmitic acids,have been shown to increase the pro-apoptotic activity of human cancer cells and inhibit cell migration and angiogenesis.This review focuses on the biological synthesis and pharmacology of endogenous cannabinoids for the enhancement of cancer cell apoptosis,which have potential as a novel therapy for GBM.展开更多
OBJECTIVE:To investigate how Chaiyuwendan decoction(CWD)affects endocannabinoid levels in the adipose tissue of depressed rats.METHODS:Twenty-four male Sprague-Dawley rats were randomly divided into four groups with s...OBJECTIVE:To investigate how Chaiyuwendan decoction(CWD)affects endocannabinoid levels in the adipose tissue of depressed rats.METHODS:Twenty-four male Sprague-Dawley rats were randomly divided into four groups with six rats in each.One group was randomly selected as the control group.The remaining three groups were subjected to chronic stress to induce depression.Groups were randomly assigned as a model group,CWD group,and amitriptyline group.CWD was given to the CWD group once a day from the second day of modeling.The amitriptyline group was administered amitriptyline intragastrically(10 mg/kg)once a day.After treatment for 21 days,body weight and fat weight were measured and the levels of N-arachidonoylethanolamine(AEA),2-arachidonoylglycerol(2-AG),and N-palmitoylethanolamine(PEA)in adipose tissue were determined with liquid chromatography-mass spectrometry.RESULTS:Compared with the control group,body weight,fat weight,AEA,and PEA were significantly lower,and 2-AG was higher,in the model group(P<0.05,P<0.01).Compared with the model group,body weight,fat weight,the AEA,and PEA levels were significantly higher,and 2-AG level was significantly lower in the CWD group(P<0.05).However,the levels did not differ significantly between the CWD group and the amitriptyline group.CONCLUSION:CWD could regulate the levels of AEA,2-AG,and PEA in rats with depression induced by chronic stress.展开更多
Neurodegenerative diseases are characterized by progressive degeneration and/or death of neuronal cells and results in a wide array of cognitive impairments and other serious neurological defects. The signaling pathwa...Neurodegenerative diseases are characterized by progressive degeneration and/or death of neuronal cells and results in a wide array of cognitive impairments and other serious neurological defects. The signaling pathways and definite cause underlying the development of neurodegenerative nerve diseases have not been well defined. There is evidence of mechanisms within the endocannabinoid system that may suggest important pathways involved the progression of neurodegenerative diseases as well as some cancers. The endocannabinoid system is an endogenous ubiquitous neuromodulatory system that plays a critical in the development of the central nervous system (CNS), synaptic plasticity, as well as other primary neuronal functions. The recent identification of various cannabinoid receptors and their endogenous lipid ligands has generated an interest and significant increase in research of the endocannabinoid system and its role in human health and diseases. The Endocannabinoid system possesses essential endogenous receptors—cannabinoid receptors type 1 (CB1R) and type 2 (CB2R)—which are involved in mechanisms that contribute to the progression of neurodegenerative diseases and some cancers. In this review, we discuss the role of the endocannabinoid system in various neurodegenerative diseases as well as some cancers, and its promise as a targeted pharmacological therapy for patients of neurodegenerative diseases.展开更多
Morinda citrifolia (noni) fruit juice has been shown to have a wide variety of potential health benefits in human clinical trials. It may also influence the endocannabinoid system of the body. Since the main ingredien...Morinda citrifolia (noni) fruit juice has been shown to have a wide variety of potential health benefits in human clinical trials. It may also influence the endocannabinoid system of the body. Since the main ingredient of the product studied in these clinical trials was juice made from noni fruit puree from French Polynesia, it was evaluated for its ability to inhibit the two major endocannabinoid degradation enzymes in vitro. Noni fruit juice inhibited both fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL) in a concentration-dependent manner, suggesting that it may help maintain anandamide and 2-arachidonoylglycerol levels. Samples of the puree were also analyzed for the presence of characteristic phytochemical markers of authentic noni fruit such as scopoletin, rutin, quercetin, deacetylasperulosidic acid and asperulosidic acid, all of which were present. Also present was scandoside, which is reported for the first time as being identified in noni fruit or its juice. Some of these compounds may contribute to the FAAH and MAGL inhibiting activity of noni juice. These results reveal another set of mechanisms by which noni juice possibly supports mental health, maintains joint health, relieves discomfort and modulates the immune system.展开更多
Cannabis has a detrimental impact on the developing nervous system. Therefore, regular consumption of cannabis by pregnant and lactating woman poses a potential risk to neuronal growth in fetuses and infants. Indeed, ...Cannabis has a detrimental impact on the developing nervous system. Therefore, regular consumption of cannabis by pregnant and lactating woman poses a potential risk to neuronal growth in fetuses and infants. Indeed, endogenous cannabis-like molecules called endocannabinoids regulate many physiological processes, including neurogenesis, axon guidance, and synaptic plasticity through CB1 receptors. To investigate the physiological role of CB1 receptors on peripheral sensory nerve growth, the endocannabinoid 2-arachidonoyl glycerol was added to cultured chick dorsal root ganglion neurons. This compound inhibited neurite elongation and induced growth cone collapse in a dose- and time-dependent manner. These data suggest that caution should be exercised regarding maternal cannabis use during pregnancy. Because ectopic sprouting and abnormal neuronal network connections are considered to be a cause of neuropathic pain, our current data imply an additional role of endocannabinoids as inhibitors of the formation of pain-maintenance networks.展开更多
Alcohol is a well-recognized teratogen that can causevariable physical and behavioral effects on the fetus.Alcohol use and abuse during pregnancy is one of the major health and societal problems and has been linked to...Alcohol is a well-recognized teratogen that can causevariable physical and behavioral effects on the fetus.Alcohol use and abuse during pregnancy is one of the major health and societal problems and has been linked to a wide range of birth defects in the offspring collectively termed as fetal alcohol spectrum disorder(FASD). The severity of abnormalities may depend on a number of factors that include the amount, the frequency, the period during gestation and the route of alcohol administration. The current knowledge about the neurobiological basis of FASD is limited. However, recent studies have suggested that the membrane-derived lipids especially bioactive endogenous cannabinoids(eCB)such as arachidonyl ethanolamide and 2-arachidonyl glycerol resulting from alcohol exposure, may play a significant role in modulating neurophysiological and neurobehavioral effects in chronic alcohol exposed adult animals. Based on these findings and on reported studies on the role of eCB signaling in neurodevelopment and behavior, it is speculated that the eC B signaling may play a critical role in fetal alcohol syndrome and FASD-related behavioral effects. The current discussion will touch upon some of the mechanistic explanations about the role of eCB signaling system in FASD and provide further guidance for future direction.展开更多
The cannabinoid （CB） receptors, endocannabinoids （eCB） and their synthesizing and catabolizing enzymes and the proteins involved in their transport, constitute what is now recognized as the eCB system. The eCBs ar...The cannabinoid （CB） receptors, endocannabinoids （eCB） and their synthesizing and catabolizing enzymes and the proteins involved in their transport, constitute what is now recognized as the eCB system. The eCBs are a class of lipids that have been identifed as retro-grade messengers and produce their effects via presyn-aptic CB receptors. The major function of the eCBs has been suggested to be that of modulating the release of several neurotransmitters implicated in a number of biological functions that include reward and reinforce-ment. There is now significant evidence to suggest that the eCB system plays an important role in the development of alcohol tolerance, dependence and relapse. Recent studies suggest that the pharmacological manipulation of the eCB system has the potential not only to block the direct reinforcing properties of alcohol but also alleviate behavioral abnormalities associated with relapse. There is also accumulating evidence that points to the possible utility of the eCB system targeted drugs in the treatment of alcoholism-related behavioral disorders. The agents that block CB1 receptor function or inhibit the synthesis of eCBs are attractive candidate drugs that need to be explored. Further understanding of the role of the eCB system in molecular mechanism/s that underlies alcoholism-related behaviors should lead to a better treatment of this devastating disorder.展开更多
Background: The endocannabinoid (EC) system is well characterized in the central nervous system but scarcely studied in peripheral organs. In this paper, we newly identify the effect of the EC anandamide (AEA) upon re...Background: The endocannabinoid (EC) system is well characterized in the central nervous system but scarcely studied in peripheral organs. In this paper, we newly identify the effect of the EC anandamide (AEA) upon renal proximal tubule cells. Methods: Measurement of lactate dehydrogenase (LDH) release after treatment of primary renal proximal tubule cells (RPTEC) and renal carcinoma cell line (Caki-1) with AEA, arachidonic acid (AA), ethanolamide (EtAm), EC receptor CB1 antagonist (AM251), CB2 receptor antagonist (SR144528), TRPV1 receptor antagonist (capsazepine), degradation enzyme fatty acid amide hydrolase (FAAH) antagonist (URB597), antioxidants GSH-EE;Trolox, GSH depletor BSO, membrane cholesterol depletor (MCD), apoptosis inhibitor zVAD, necroptosis inhibitor Nec-1 or ferroptosis inhibitor Fer-1. Western blot and qRT-PCR analysis plus determination of reactive oxygen species (ROS) via H2-DCFDA were performed. Histology for EC enzymes, N-acetylphosphatidylethanolamine-hydrolyzing phospholipase D (NAPE-PLD) and FAAH, as well as the determination of physiological levels of ECs in human and rat renal tissue via liquid chromatography were conducted. Results: AEA both dose- and time-dependently induces cell death in RPTEC and Caki-1 within hours, characterized by cell blebbing, not influenced by blocking the described EC receptors by AM251, SR144528, capsazepine or FAAH by URB597 or MCD. Cell death is mediated via ROS. There is no difference found in the histology of the enzymes FAAH and NAPE-PLD in human renal tissue with interstitial nephritis. Blocking of apoptotic, necroptotic or ferroptotic cell death does not lead to a reduction in LDH release in vitro. Conclusion: The endocannabinoid anandamide induces cell death in renal proximal tubule cell in a time- and dose-dependent manner. This pathway is mediated via ROS and is independent of cannabinoid receptors, membrane cholesterol or FAAH activity.展开更多
基金supported by the Key Program of the National Natural Science Foundation of China(82130122)the National Natural Science Foundation of China(81973949).
文摘Pain is a subjective and unpleasant sensation that significantly impacts the daily lives of individuals.Chronic pain represents one of the most challenging public health issues,and ensuring effective pain management is a fundamental right of individuals and a sacred duty of healthcare providers.Cannabis,one of the earliest recognized medicinal plants,contains cannabinoids,which are non-opioid substances that modulate nociceptive responses.Electroacupuncture(EA),characterized by its low-risk and well-tolerated nature,is pivotal in pain management.The endocannabinoid system consists of endocannabinoids,cannabinoid receptors,and enzymes involved in endocannabinoid synthesis,degradation,and transport.Recently,the role of the endocannabinoid system in pain development and EA analgesia has attracted considerable research attention.Studies have highlighted the role of the endocannabinoid system in various types of pain,including inflammatory pain,neuropathic pain,and cancer-related pain,as well as in EA analgesia.This study aims to review the mechanisms of endocannabinoid system involvement in pain modulation and EA analgesia to provide insights to inform clinical approaches to pain management.
基金funded by the Michigan Animal Agricultural Alliance competitive project#AA-23–0014United States Department of Agriculture National Institute of Food and Agriculture(USDA)projects#2019–67015-29443 and#2021–67037-34657+2 种基金the United States-Israel Binational Agricultural Research Development(BARD)Fund project#IS-5167–19the Office for the Associate Dean of Research at Michigan State Universitysupported by USDA National Institute of Food and Agriculture’s Agriculture and Food Research Initiative Educational Workforce and Development Program award#2024-67011-42947。
文摘Background As cows transition from pregnancy to lactation,free fatty acids(FFA)are mobilized from adipose tissues(AT)through lipolysis to counter energy deficits.In clinically healthy cows,lipolysis intensity is reduced throughout lactation;however,if F FA release exceeds tissue demands or the liver's metabolic capacity,lipid byproducts accumulate,incre a sing cows'risk of metabolic and infectious disease.Endocannabinoids(eCBs)and their congeners,N-acylethanolamines(NAEs),are lipid-based compounds that modulate metabolism and inflammation.Their synthesis and release depend upon the availability of FFA precursors and the abundance of synthesizing and degrading enzymes and transporters.Therefore,we hypothesized that eCB production and transcription of endocannabinoid system components are modulated by lipolysis pathways in adipocytes.To test this hypothesise we stimulated canonical(isoproterenol,1μmol/L;ISO)and inflammatory(lipopolysaccharide,1μg/m L;LPS)lipolysis pathways in adipocytes isolated from the AT of 5 Holstein dairy cows.Following,we assessed lipolysis intensity,adipocytes'release of eCBs,and transcription of endocannabinoid system components.Results We found that ISO and LPS stimulated lipolysis at comparable intensities.Exposure to either treatment tended to elevate the release of eCBs and NAEs by cultured adipocytes;however,specific eCBs and NAEs and the transcriptional profiles differed by treatment.On one hand,ISO enhanced adipocytes'release of 2-arachidonoylglycerol(2-AG)but reduced NAE production.Klotably,ISO enhanced the cells"expression of enzymes associated with 2-AG biosynthesis(INPP5F,GDPD5,GPAT4),transport(CD36),and adipogenesis(PPARG).Conversely,LPS enhanced adipocytes'synthesis and release of N-arachidonoylethanolamide(AEA).This change coincided with enhanced transcription of the NAE-biosynthesizing enzyme,PTPN22,and adipocytes'transcription of genes related to eCB degradation(PTGS2,MGLL,CYP27B1).Furthermore,LPS enhanced adipocytes'transcription of eCB and NAE transporters(HSPA1A,SCP2)and the expression of the anti-adipogenic ion channel,TRPV3.Conclusions Our data provide evidence for distinct modulatory roles of canonical and inflammatory lipolysis pathways over eCB release and transcriptional regulation of biosynthesis,degradation,transport,and ECS signaling in cows'adipocytes.Based on our findings,we conclude that,within adipocytes,eCB production and ECS component expression are,at least in part,mediated by lipolysis in a pathway-dependent manner.These findings contribute to a deeper understanding of the molecular mechanisms underlying metabolic regulation in dairy cows'AT,with potential implications for prevention and treatment of inflammatory and metabolic disorders.
基金financially supported by the Chief Scientist of the Ministry of Agriculture,grant number 20-04-0015,Rishon Lezion,Israel。
文摘Background The placenta plays a crucial role in supporting and influencing fetal development.We compared the effects of prepartum supplementation with omega-3(n-3)fatty acid(FA)sources,flaxseed oil(FLX)and fish oil(FO),on the expression of genes and proteins related to lipid metabolism,inflammation,oxidative stress,and the endocannabinoid system(ECS)in the expelled placenta,as well as on FA profile and inflammatory response of neonates.Late-pregnant Holstein dairy cows were supplemented with saturated fat(CTL),FLX,or FO.Placental cotyledons(n=5)were collected immediately after expulsion,and extracted RNA and proteins were analyzed by RTPCR and proteomic analysis.Neonatal blood was assessed for FA composition and concentrations of inflammatory markers.Results FO increased the gene expression of fatty acid binding protein 4(FABP4),interleukin 10(IL-10),catalase(CAT),cannabinoid receptor 1(CNR1),and cannabinoid receptor 2(CNR2)compared with CTL placenta.Gene expression of ECS-enzyme FA-amide hydrolase(FAAH)was lower in FLX and FO than in CTL.Proteomic analysis identified 3,974 proteins;of these,51–59 were differentially abundant between treatments(P≤0.05,|fold change|≥1.5).Top canonical pathways enriched in FLX vs.CTL and in FO vs.CTL were triglyceride metabolism and inflammatory processes.Both n-3 FA increased the placental abundance of FA binding proteins(FABPs)3 and 7.The abundance of CNR1 cannabinoid-receptor-interacting-protein-1(CNRIP1)was reduced in FO vs.FLX.In silico modeling affirmed that bovine FABPs bind to endocannabinoids.The FLX increased the abundance of inflammatory CD44-antigen and secreted-phosphoprotein-1,whereas prostaglandin-endoperoxide synthase 2 was decreased in FO vs.CTL placenta.Maternal FO enriched neonatal plasma with n-3 FAs,and both FLX and FO reduced interleukin-6 concentrations compared with CTL.Conclusion Maternal n-3 FA from FLX and FO differentially affected the bovine placenta;both enhanced lipid metabolism and modulated oxidative stress,however,FO increased some transcriptional ECS components,possibly related to the increased FABPs.Maternal FO induced a unique balance of pro-and anti-inflammatory components in the placenta.Taken together,different sources of n-3 FA during late pregnancy enhanced placental immune and metabolic processes,which may affect the neonatal immune system.
基金supported in part by grants from the National Institute on Aging and National Eye Institute[EY030747(3R01EY030747-02S2)and EY031248(3R01EY03124802S1)]of the National Institutes of Health(to PK)Additional support by the Felix and Carmen Sabates Missouri Endowed Chair in Vision Researchthe Vision Research Foundation of Kansas City。
文摘Cannabinoids are lipophilic substances derived from Cannabis sativa that can exert a variety of effects in the human body.They have been studied in cellular and animal models as well as in human clinical trials for their therapeutic benefits in several human diseases.Some of these include central nervous system(CNS)diseases and dysfunctions such as forms of epilepsy,multiple sclerosis,Parkinson’s disease,pain and neuropsychiatric disorders.In addition,the endogenously produced cannabinoid lipids,endocannabinoids,are critical for normal CNS function,and if controlled or modified,may represent an additional therapeutic avenue for CNS diseases.This review discusses in vitro cellular,ex vivo tissue and in vivo animal model studies on cannabinoids and their utility as therapeutics in multiple CNS pathologies.In addition,the review provides an overview on the use of cannabinoids in human clinical trials for a variety of CNS diseases.Cannabinoids and endocannabinoids hold promise for use as disease modifiers and therapeutic agents for the prevention or treatment of neurodegenerative diseases and neurological disorders.
基金This project was funded by the US-Israel Binational Agricultural Research and Development Fund(Grant IS-5167-19)MNS was supported in part by the USDA National Institute for Food and Agriculture(Washington,DC,USA)competitive project 2019-67015-29443.
文摘To sustain the nutrient demands of rapid fetal growth,parturition,and milk synthesis,periparturient dairy cows mobilize adipose tissue fatty acid stores through lipolysis.This process induces an inflammatory response within AT that is resolved as lactation progresses;however,excessive and protracted lipolysis compounds the risk for metabolic and inflammatory diseases.The suppression of lipolytic action and inflammation,along with amplification of adipogenesis and lipogenesis,serve as prospective therapeutic targets for improving the health of periparturient dairy cows.Generally,the activation of cannabinoid receptors by endocannabinoids enhances adipogenesis and lipogenesis,suppresses lipolysis,and increases appetite in mammals.These biological effects of activating the endocannabinoid system open the possibility of harnessing the endocannabinoid system through nutritional intervention in dairy herds as a potential tool to improve dairy cows’health,although much is still to be revealed in this context.This review summarizes the current knowledge surrounding the components of the endocannabinoid system,elaborates on the metabolic effects of its activation,and explores the potential to modulate its activity in periparturient dairy cows.
基金supported by the Study Grant (2007) for Medical Students of Second Military Medical University,China
文摘Being a great threaten for human health, obesity has become a pandemic chronic disease. There have been several therapeutic treatments for this social health issue, including diet and exercise therapy, medication and surgery, among which the diet is still the most common way. However, none of these therapeutic measures available is ideal, making it necessary to find an effective medical treatment. The endocannabinoid system, which is well known for its contributions in certain mental processes such as relaxation, amelioration of pain and anxiety, and sedation initiation, has been recently reported to play an essential role in regulating appetite and metabolism to maintain energy balance, leading to the belief that endocannabinoid system is closely related to obesity. This new discovery deepens our understanding of obesity, and provides us with a new direction for clinical obesity treatment. Rimonabant is an antagonist for CB1, and has entered the market in some countries. However, although effective as an anti-obesity drug, rimonabant also causes obviously adverse side-effects, thus is being doubted and denied for medical usage.
文摘The endocannabinoid system(ECS),particularly its signaling pathways and ligands,has garnered considerable interest in recent years.Along with clinical work investigating the ECS’functions,including its role in the development of neurological and inflammatory conditions,much research has focused on developing analytical protocols enabling the precise monitoring of the levels and metabolism of the most potent ECS ligands:exogenous phytocannabinoids(PCs)and endogenous cannabinoids(endocannabinoids,ECs).Solid-phase microextraction(SPME)is an advanced,non-exhaustive sample-preparation technique that facilitates the precise and efficient isolation of trace amounts of analytes,thus making it appealing for the analysis of PCs and ECs in complex matrices of plant and animal/human origin.In this paper,we review recent forensic medicine and toxicological studies wherein SPME has been applied to monitor levels of PCs and ECs in complex matrices,determine their effects on organism physiology,and assess their role in the development of several diseases.
文摘Diabetic complications,chiefly seen in long-term situations,are persistently deleterious to a large extent,requiring multi-factorial risk reduction strategies beyond glycemic control.Diabetic cardiomyopathy is one of the most common deleterious diabetic complications,being the leading cause of mortality among diabetic patients.The mechanisms of diabetic cardiomyopathy are multi-factorial,involving increased oxidative stress,accumulation of advanced glycation end products(AGEs),activation of various pro-inflammatory and cell death signaling pathways,and changes in the composition of extracellular matrix with enhanced cardiac fibrosis.The novel lipid signaling system,the endocannabinoid system,has been implicated in the pathogenesis of diabetes and its complications through its two main receptors:Cannabinoid receptor type 1 and cannabinoid receptor type 2,alongside other components.However,the role of the endocannabinoid system in diabetic cardiomyopathy has not been fully investigated.This review aims to elucidate the possible mechanisms through which cannabinoids and the endocannabinoid system could interact with the pathogenesis and the development of diabetic cardiomyopathy.These mechanisms include oxidative/nitrative stress,inflammation,accumulation of AGEs,cardiac remodeling,and autophagy.A better understanding of the role of cannabinoids and the endocannabinoid system in diabetic cardiomyopathy may provide novel strategies to manipulate such a serious diabetic complication.
基金Research Grant No.IS-5167–19 from BARDThe United States—Israel Binational Agricultural Research and Development Fund(Rishon Lezion,Israel)to J.T,G.A.C and M.ZJ.R.D received a postdoc scholarship from the Israeli Ministry of Foreign Affairs。
文摘Background:Dietary supplementation of omega-3 fatty acids can reduce the activation of the endocannabinoid system(ECS)by decreasing the availability of arachidonic acid,thus lowering endocannabinoids(e CBs)levels.The ECS is a modulator of energy metabolism,stress response and inflammation in mammals,yet there is little information on the roles of the ECS in transition dairy cows.During the periparturient period,the adipose tissue and liver are the main metabolic organs that participate in the adaptations of dairy cows to onset of lactation;however,exceeded adipose tissue lipolysis and accumulation of lipids in the liver have adverse effects on cows'physiology.Here we aimed to examine whether omega-3 supplementation during the transition period will modulate ECS activation and affect metabolic and inflammatory indices in postpartum dairy cows,by supplementing twenty-eight transition Holstein dairy cows with either saturated fat(CTL)or encapsulated flaxseed oil(FLX).Components of the ECS,metabolic and inflammatory markers were measured in blood,liver,and subcutaneous adipose tissue.Results:FLX supplementation reduced feed intake by 8.1%(P<0.01)and reduced plasma levels of arachidonic acid(by 44.2%;P=0.02)and anandamide(by 49.7%;P=0.03)postpartum compared to CTL.The m RNA transcription levels of the cannabinoid receptor 1(CNR1/CB1)tended to be lower(2.5 folds)in white blood cells of FLX than in CTL(P=0.10),and protein abundance of ECS enzyme monoacylglycerol lipase was higher in peripheral blood mononuclear cells of FLX than in CTL(P=0.04).In adipose tissue,palmitoylethanolamide levels were lower in FLX than in CTL(by 61.5%;P=0.02),relative m RNA transcription of lipogenic genes were higher,and the protein abundance of cannabinoid receptor 2(P=0.08)and monoacylglycerol lipase(P=0.10)tended to be higher in FLX compared to CTL.Hepatic 2-arachidonoylglycerol tended to be higher(by 73.1%;P=0.07),and interlukin-6 m RNA transcription level was 1.5 folds lower in liver of FLX than in CTL(P=0.03).Conclusions:Nutritional supplementation of omega-3 fatty acids seems to partly modulate ECS activation,which could be related to lower feed intake.The altered ECS components in blood,adipose tissue and liver are associated with moderate modulations in lipid metabolism in the adipose and inflammation in liver of peripartum dairy cows.
基金supported by National Institutes of Health grants NS076815
文摘Humans have been using Cannabis and its extracts for a few thousand years as a medicinal and recreational drug. How- ever, the chemical component in Cannabis sativa, △9-tet- rahydrocannabinol （△9-THC）, an exogenous cannabinoid, remained unknown until it was isolated and identified as the main psychoactive ingredient （Gaoni and Mechoulam, 1964）.
文摘Globally,it is evident that glioblastoma multiforme(GBM)is an aggressive malignant cancer with a high mortality rate and no effective treatment options.Glioblastoma is classified as the stage-four progression of a glioma tumor,and its diagnosis results in a shortened life expectancy.Treatment options for GBM include chemotherapy,immunotherapy,surgical intervention,and conventional pharmacotherapy;however,at best,they extend the patient’s life by a maximum of 5 years.GBMs are considered incurable due to their high recurrence rate,despite various aggressive therapeutic approaches which can have many serious adverse effects.Ceramides,classified as endocannabinoids,offer a promising novel therapeutic approach for GBM.Endocannabinoids may enhance the apoptosis of GBM cells but have no effect on normal healthy neural cells.Cannabinoids promote atypical protein kinase C,deactivate fatty acid amide hydrolase enzymes,and activate transient receptor potential vanilloid 1(TRPV1)and TRPV2 to induce pro-apoptotic signaling pathways without increasing endogenous cannabinoids.In previous in vivo studies,endocannabinoids,chemically classified as amide formations of oleic and palmitic acids,have been shown to increase the pro-apoptotic activity of human cancer cells and inhibit cell migration and angiogenesis.This review focuses on the biological synthesis and pharmacology of endogenous cannabinoids for the enhancement of cancer cell apoptosis,which have potential as a novel therapy for GBM.
基金Supported by the Natural Science Fund of Fujian(No.13141240)a Xiamen Science and Technology Key Program Grant(No.3502Z20100006)the Undergraduate Innovative Experiment Program(No.DC2013271)
文摘OBJECTIVE:To investigate how Chaiyuwendan decoction(CWD)affects endocannabinoid levels in the adipose tissue of depressed rats.METHODS:Twenty-four male Sprague-Dawley rats were randomly divided into four groups with six rats in each.One group was randomly selected as the control group.The remaining three groups were subjected to chronic stress to induce depression.Groups were randomly assigned as a model group,CWD group,and amitriptyline group.CWD was given to the CWD group once a day from the second day of modeling.The amitriptyline group was administered amitriptyline intragastrically(10 mg/kg)once a day.After treatment for 21 days,body weight and fat weight were measured and the levels of N-arachidonoylethanolamine(AEA),2-arachidonoylglycerol(2-AG),and N-palmitoylethanolamine(PEA)in adipose tissue were determined with liquid chromatography-mass spectrometry.RESULTS:Compared with the control group,body weight,fat weight,AEA,and PEA were significantly lower,and 2-AG was higher,in the model group(P<0.05,P<0.01).Compared with the model group,body weight,fat weight,the AEA,and PEA levels were significantly higher,and 2-AG level was significantly lower in the CWD group(P<0.05).However,the levels did not differ significantly between the CWD group and the amitriptyline group.CONCLUSION:CWD could regulate the levels of AEA,2-AG,and PEA in rats with depression induced by chronic stress.
文摘Neurodegenerative diseases are characterized by progressive degeneration and/or death of neuronal cells and results in a wide array of cognitive impairments and other serious neurological defects. The signaling pathways and definite cause underlying the development of neurodegenerative nerve diseases have not been well defined. There is evidence of mechanisms within the endocannabinoid system that may suggest important pathways involved the progression of neurodegenerative diseases as well as some cancers. The endocannabinoid system is an endogenous ubiquitous neuromodulatory system that plays a critical in the development of the central nervous system (CNS), synaptic plasticity, as well as other primary neuronal functions. The recent identification of various cannabinoid receptors and their endogenous lipid ligands has generated an interest and significant increase in research of the endocannabinoid system and its role in human health and diseases. The Endocannabinoid system possesses essential endogenous receptors—cannabinoid receptors type 1 (CB1R) and type 2 (CB2R)—which are involved in mechanisms that contribute to the progression of neurodegenerative diseases and some cancers. In this review, we discuss the role of the endocannabinoid system in various neurodegenerative diseases as well as some cancers, and its promise as a targeted pharmacological therapy for patients of neurodegenerative diseases.
文摘Morinda citrifolia (noni) fruit juice has been shown to have a wide variety of potential health benefits in human clinical trials. It may also influence the endocannabinoid system of the body. Since the main ingredient of the product studied in these clinical trials was juice made from noni fruit puree from French Polynesia, it was evaluated for its ability to inhibit the two major endocannabinoid degradation enzymes in vitro. Noni fruit juice inhibited both fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL) in a concentration-dependent manner, suggesting that it may help maintain anandamide and 2-arachidonoylglycerol levels. Samples of the puree were also analyzed for the presence of characteristic phytochemical markers of authentic noni fruit such as scopoletin, rutin, quercetin, deacetylasperulosidic acid and asperulosidic acid, all of which were present. Also present was scandoside, which is reported for the first time as being identified in noni fruit or its juice. Some of these compounds may contribute to the FAAH and MAGL inhibiting activity of noni juice. These results reveal another set of mechanisms by which noni juice possibly supports mental health, maintains joint health, relieves discomfort and modulates the immune system.
文摘Cannabis has a detrimental impact on the developing nervous system. Therefore, regular consumption of cannabis by pregnant and lactating woman poses a potential risk to neuronal growth in fetuses and infants. Indeed, endogenous cannabis-like molecules called endocannabinoids regulate many physiological processes, including neurogenesis, axon guidance, and synaptic plasticity through CB1 receptors. To investigate the physiological role of CB1 receptors on peripheral sensory nerve growth, the endocannabinoid 2-arachidonoyl glycerol was added to cultured chick dorsal root ganglion neurons. This compound inhibited neurite elongation and induced growth cone collapse in a dose- and time-dependent manner. These data suggest that caution should be exercised regarding maternal cannabis use during pregnancy. Because ectopic sprouting and abnormal neuronal network connections are considered to be a cause of neuropathic pain, our current data imply an additional role of endocannabinoids as inhibitors of the formation of pain-maintenance networks.
文摘Alcohol is a well-recognized teratogen that can causevariable physical and behavioral effects on the fetus.Alcohol use and abuse during pregnancy is one of the major health and societal problems and has been linked to a wide range of birth defects in the offspring collectively termed as fetal alcohol spectrum disorder(FASD). The severity of abnormalities may depend on a number of factors that include the amount, the frequency, the period during gestation and the route of alcohol administration. The current knowledge about the neurobiological basis of FASD is limited. However, recent studies have suggested that the membrane-derived lipids especially bioactive endogenous cannabinoids(eCB)such as arachidonyl ethanolamide and 2-arachidonyl glycerol resulting from alcohol exposure, may play a significant role in modulating neurophysiological and neurobehavioral effects in chronic alcohol exposed adult animals. Based on these findings and on reported studies on the role of eCB signaling in neurodevelopment and behavior, it is speculated that the eC B signaling may play a critical role in fetal alcohol syndrome and FASD-related behavioral effects. The current discussion will touch upon some of the mechanistic explanations about the role of eCB signaling system in FASD and provide further guidance for future direction.
基金Supported by Funds from the National Institute of Health,Bethesda,United StatesAmerican Foundation for SuicidePrevention
文摘The cannabinoid （CB） receptors, endocannabinoids （eCB） and their synthesizing and catabolizing enzymes and the proteins involved in their transport, constitute what is now recognized as the eCB system. The eCBs are a class of lipids that have been identifed as retro-grade messengers and produce their effects via presyn-aptic CB receptors. The major function of the eCBs has been suggested to be that of modulating the release of several neurotransmitters implicated in a number of biological functions that include reward and reinforce-ment. There is now significant evidence to suggest that the eCB system plays an important role in the development of alcohol tolerance, dependence and relapse. Recent studies suggest that the pharmacological manipulation of the eCB system has the potential not only to block the direct reinforcing properties of alcohol but also alleviate behavioral abnormalities associated with relapse. There is also accumulating evidence that points to the possible utility of the eCB system targeted drugs in the treatment of alcoholism-related behavioral disorders. The agents that block CB1 receptor function or inhibit the synthesis of eCBs are attractive candidate drugs that need to be explored. Further understanding of the role of the eCB system in molecular mechanism/s that underlies alcoholism-related behaviors should lead to a better treatment of this devastating disorder.
基金supported by Koeln Fortune Program/Faculty of Medicine,University of Cologne and excellence cluster initiative supported by University of Cologne and DFG.
文摘Background: The endocannabinoid (EC) system is well characterized in the central nervous system but scarcely studied in peripheral organs. In this paper, we newly identify the effect of the EC anandamide (AEA) upon renal proximal tubule cells. Methods: Measurement of lactate dehydrogenase (LDH) release after treatment of primary renal proximal tubule cells (RPTEC) and renal carcinoma cell line (Caki-1) with AEA, arachidonic acid (AA), ethanolamide (EtAm), EC receptor CB1 antagonist (AM251), CB2 receptor antagonist (SR144528), TRPV1 receptor antagonist (capsazepine), degradation enzyme fatty acid amide hydrolase (FAAH) antagonist (URB597), antioxidants GSH-EE;Trolox, GSH depletor BSO, membrane cholesterol depletor (MCD), apoptosis inhibitor zVAD, necroptosis inhibitor Nec-1 or ferroptosis inhibitor Fer-1. Western blot and qRT-PCR analysis plus determination of reactive oxygen species (ROS) via H2-DCFDA were performed. Histology for EC enzymes, N-acetylphosphatidylethanolamine-hydrolyzing phospholipase D (NAPE-PLD) and FAAH, as well as the determination of physiological levels of ECs in human and rat renal tissue via liquid chromatography were conducted. Results: AEA both dose- and time-dependently induces cell death in RPTEC and Caki-1 within hours, characterized by cell blebbing, not influenced by blocking the described EC receptors by AM251, SR144528, capsazepine or FAAH by URB597 or MCD. Cell death is mediated via ROS. There is no difference found in the histology of the enzymes FAAH and NAPE-PLD in human renal tissue with interstitial nephritis. Blocking of apoptotic, necroptotic or ferroptotic cell death does not lead to a reduction in LDH release in vitro. Conclusion: The endocannabinoid anandamide induces cell death in renal proximal tubule cell in a time- and dose-dependent manner. This pathway is mediated via ROS and is independent of cannabinoid receptors, membrane cholesterol or FAAH activity.
