Objective To investigate the structural changes of rat thoracic aorta and changes in expression levels of Bmal1 and cyclins in thoracic aorta endothelial cells following heat stress.Methods Twenty male SD rats were ra...Objective To investigate the structural changes of rat thoracic aorta and changes in expression levels of Bmal1 and cyclins in thoracic aorta endothelial cells following heat stress.Methods Twenty male SD rats were randomized equally into control group and heat stress group.After exposure to 32℃for 2 weeks in the latter group,the rats were examined for histopathological changes and Bmal1 expression in the thoracic aorta using HE staining and immunohistochemistry.In the cell experiments,cultured rat thoracic aortic endothelial cells(RTAECs)were incubated at 40℃for 12 h with or without prior transfection with a Bmal1-specific small interfering RNA(si-Bmal1)or a negative sequence.In both rat thoracic aorta and RTAECs,the expressions of Bmal1,the cell cycle proteins CDK1,CDK4,CDK6,and cyclin B1,and apoptosis-related proteins Bax and Bcl-2 were detected using Western blotting.TUNEL staining was used to detect cell apoptosis in rat thoracic aorta,and the changes in cell cycle distribution and apoptosis in RTAECs were analyzed with flow cytometry.Results Compared with the control rats,the rats exposed to heat stress showed significantly increased blood pressures and lowered heart rate with elastic fiber disruption and increased expressions of Bmal1,cyclin B1 and CDK1 in the thoracic aorta(P<0.05).In cultured RTAECs,heat stress caused significant increase of Bmal1,cyclin B1 and CDK1 protein expression levels,which were obviously lowered in cells with prior si-Bmal1 transfection.Bmal1 knockdown also inhibited heat stress-induced increase of apoptosis in RTAECs as evidenced by decreased expression of Bax and increased expression of Bcl-2.Conclusion Heat stress upregulates Bmal1 expression and causes alterations in expressions of cyclins to trigger apoptosis of rat thoracic aorta endothelial cells,which can be partly alleviated by suppressing Bmal1 expression.展开更多
The circadian clock is an important internal time regulatory system for a range of physiological and behavioral rhythms within living organisms.Testosterone,as one of the most critical sex hormones,is essential for th...The circadian clock is an important internal time regulatory system for a range of physiological and behavioral rhythms within living organisms.Testosterone,as one of the most critical sex hormones,is essential for the development of the reproductive system,maintenance of reproductive function,and the overall health of males.The secretion of testosterone in mammals is characterized by distinct circadian rhythms and is closely associated with the regulation of circadian clock genes.Here we review the central and peripheral regulatory mechanisms underlying the influence of circadian clock genes upon testosterone synthesis.We also examined the specific effects of these genes on the occurrence,development,and treatment of common male diseases,including late-onset hypogonadism,erectile dysfunction,male infertility,and prostate cancer.展开更多
Clock genes create a complicated molecular time-keeping system consisting of multiple positive and negative feedback loops at transcriptional and translational levels. This circadian system coordinates and regulates m...Clock genes create a complicated molecular time-keeping system consisting of multiple positive and negative feedback loops at transcriptional and translational levels. This circadian system coordinates and regulates multiple cellular procedures implicated in cancer development such as metabolism, cell cycle and DNA damage response. Recent data support that molecules such as CLOCK1, BMAL1 and PER and CRY proteins have various effects on c-Myc/p21 and Wnt/β-catenin pathways and influence multiple steps of DNA damage response playing a critical role in the preservation of genomic integrity in normal and cancer cells. Notably, all these events have already been related to the development and progression of colorectal cancer (CRC). Recent data highlight critical correlations between clock genes’ expression and pathogenesis, progression, aggressiveness and prognosis of CRC. Increased expression of positive regulators of this circadian system such as BMAL1 has been related to decrease overall survival while decreased expression of negative regulators such as PER2 and PER3 is connected with poorer differentiation, increased aggressiveness and worse prognosis. The implications of these molecules in DNA repair systems explain their involvement in the development of CRC but at the same time provide us with novel targets for modern therapeutic approaches for patients with advanced CRC.展开更多
Mammalian bone is constantly metabolized from the embryonic stage,and the maintenance of bone health depends on the dynamic balance between bone resorption and bone formation,mediated by osteoclasts and osteoblasts.It...Mammalian bone is constantly metabolized from the embryonic stage,and the maintenance of bone health depends on the dynamic balance between bone resorption and bone formation,mediated by osteoclasts and osteoblasts.It is widely recognized that circadian clock genes can regulate bone metabolism.In recent years,the regulation of bone metabolism by non-coding RNAs has become a hotspot of research.MicroRNAs can participate in bone catabolism and anabolism by targeting key factors related to bone metabolism,including circadian clock genes.However,research in this field has been conducted only in recent years and the mechanisms involved are not yet well established.Recent studies have focused on how to target circadian clock genes to treat some diseases,such as autoimmune diseases,but few have focused on the co-regulation of circadian clock genes and microRNAs in bone metabolic diseases.Therefore,in this paper we review the progress of research on the co-regulation of bone metabolism by circadian clock genes and microRNAs,aiming to provide new ideas for the prevention and treatment of bone metabolic diseases such as osteoporosis.展开更多
The Clock gene,a key molecule in circadian systems,is widely distributed in the animal kingdom. We isolated a 936-bp partial c DNA sequence of the C lock gene( Pva- clock) from the darkbarbel catfish P elteobagrus vac...The Clock gene,a key molecule in circadian systems,is widely distributed in the animal kingdom. We isolated a 936-bp partial c DNA sequence of the C lock gene( Pva- clock) from the darkbarbel catfish P elteobagrus vachelli that exhibited high identity with C lock genes of other species of fish and animals(65%–88%). The putative domains included a basic helix-loop-helix(b HLH) domain and two period-ARNT-single-minded(PAS) domains,which were also similar to those in other species of fish and animals. P va- Clock was primarily expressed in the brain,and was detected in all of the peripheral tissues sampled. Additionally,the pattern of P va- Clock expression over a 24-h period exhibited a circadian rhythm in the brain,liver and intestine,with the acrophase at zeitgeber time 21:35,23:00,and 23:23,respectively. Our results provide insight into the function of the molecular C lock of P. vachelli.展开更多
Summary： This study investigated the effects of benazepril administered in the morning or evening on the diurnal variation of renin-angiotensin-aldosterone system （RAAS） and clock genes in the kidney. The male Wist...Summary： This study investigated the effects of benazepril administered in the morning or evening on the diurnal variation of renin-angiotensin-aldosterone system （RAAS） and clock genes in the kidney. The male Wistar rat models of 5/6 subtotal nephrectomy （STNx） were established. Animals were ran- domly divided into 4 groups： sham STNx group （control）, STNx group, morning benazepril group （MB） and evening benazepril group （EB）. Benazepril was intragastfically administered at a dose of 10 mg/kg/day at 07：00 and 19：00 in the MB group and EB group respectively for 12 weeks. All the animals were synchronized to the light：dark cycle of 12：12 for 12 weeks. Systolic blood pressure （SBP）, 24-h urinary protein excretion and renal function were measured at 11 weeks. Blood samples and kidneys were collected every 4 h throughout a day to detect the expression pattern of renin activity （RA）, angio- tensin Ⅱ （Ang Ⅱ ） and aldosterone （Aid） by radioimmunoassay （RIA） and the mRNA expression profile of clock genes （bmall, dbp and per2） by real-time PCR at 12 weeks. Our results showed that no signifi- cant differences were noted in the SBP, 24-h urine protein excretion and renal function between the MB and EB groups. There were no significant differences in average Aid and RA content of a day between the MB group and EB group. The expression peak of bmall mRNA was phase-delayed by 4 to 8 h, and the diurnal variation of per2 and dbp mRNA diminished in the MB and EB groups compared with the control and STNx groups. It was concluded when the similar SBP reduction, RAAS inhibition and clock gene profile were achieved with optimal dose of benazepril, morning versus evening dosing of benazepril has the same renoprotection effects.展开更多
As the body’s internal clock,the circadian rhythm regulates the energy expenditure,appetite,and sleep.There exists a close relationship between the host circadian rhythm and gut microbiota.In this work,a circadian di...As the body’s internal clock,the circadian rhythm regulates the energy expenditure,appetite,and sleep.There exists a close relationship between the host circadian rhythm and gut microbiota.In this work,a circadian disorder mouse model induced by constant darkness(CD)was constructed to investigate the regulating effects of capsaicin(CAP)on disturbances of metabolism homeostasis and gut microbiota in the respect of circadian rhythm-related mechanisms.Our results indicated that CAP reduced weight gain induced by circadian rhythm disorder in mice by inhibiting fat accumulation in liver and adipose tissue.The rhythmic expressions of circadian clock genes and lipid-metabolism related genes in liver were also recovered by CAP.Microbial study using 16S rRNA sequencing revealed that CAP modulated the gut microbiota richness,diversity and composition,and restored diurnal oscillations of gut microbes at the phylum and family level.These results indicated that CAP could alleviate CD-induced hepatic clock gene disruption and gut microbiota dysbiosis in mice,providing theoretical basis for CAP to be used as a muti-functional ingredient with great healthpromoting effects.展开更多
Objective： Investigation of the effect of Xiaoaiping on the expression of circadian clock genes in human hepatoma HepG2 cells. Methods： Selecting the HepG2 cells in the logarithmic growth phase and assigning them to...Objective： Investigation of the effect of Xiaoaiping on the expression of circadian clock genes in human hepatoma HepG2 cells. Methods： Selecting the HepG2 cells in the logarithmic growth phase and assigning them to Xiaoaiping injection （XAP） group and control group. The two groups were treated with 75 mg/mL XAP or the same dose of normal saline. After 72 h of treatment, real-time PCR was used to detect the expression of circadian clock genes in HepG2 cells and Western Blot technology was used to detect the expression of related proteins. Results： The mRNA expression levels of PER1, NPAS2, NR1D1, and DEC1 in the XAP group was significantly higher than that in the control group （P〈 0.05）, while the mRNA expression levels of PER3, BMAL1, DEC2, and RORA were significantly lower in the XAP group than in the control group （P 〈 0.05）, and there was no significant difference between the mRNA expression levels of PER2, CRY1, CRY2, and TIM. Of course, the proteins＇ expression levels of the genes we had detected such as PERle3, CRYI-2, CLOCK, BMAL1 by Western Blot were consistent with the real-time PCR results above. Conclusion： XAP affects the expression of circadian clock genes in HepG2 cells.展开更多
Many aspects of cellular physiology display circadian(approximately 24-h)rhythms.Dysfunction of the circadian clock molecular circuitry is associated with human health derangements,including neurodegeneration,increase...Many aspects of cellular physiology display circadian(approximately 24-h)rhythms.Dysfunction of the circadian clock molecular circuitry is associated with human health derangements,including neurodegeneration,increased risk of cancer,cardiovascular diseases and the metabolic syndrome.Viruses triggering hepatitis depend tightly on the host cell synthesis machinery for their own replication,survival and spreading.Recent evidences support a link between the circadian clock circuitry and viruses’biological cycle within host cells.Currently,in vitro models for chronobiological studies of cells infected with viruses need to be implemented.The establishment of such in vitro models would be helpful to better understand the link between the clock gene machinery and viral replication/viral persistence in order to develop specifically targeted therapeutic regimens.Here we review the recent literature dealing with the interplay between hepatitis B and C viruses and clock genes.展开更多
Objective: To investigate the effects of 72 hours continuous sleep deprivation (SD) on circadian clock gene expression and oxidative stress in the rat liver. Methods: Twenty healthy male Sprague-Dawley rats were divid...Objective: To investigate the effects of 72 hours continuous sleep deprivation (SD) on circadian clock gene expression and oxidative stress in the rat liver. Methods: Twenty healthy male Sprague-Dawley rats were divided into 2 groups (n = 10 each) using a random number table: normal control group (group C), sleep deprivation group (group SD). Group SD was treated with a modified multiple platform water environment method. After 72 hours sleep deprived, the levels of AST (Aspartate transaminase ) and ALT (Alanine aminotransferase) in serum were determined. The contents of malondialdehyde (MDA), the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the liver tissue of the rats were examined in both two groups. The expression levels of CLOCK, BMAL1 and CRY1 protein in liver tissue were examined by Western blotting. Results: Compared with group C, the content of MDA, and the levels of AST and ALT in serum were significantly increased (P Conclusion: 72 hours continuous sleep deprivation can downregulate the expression of circadian clock gene and promote oxidative stress in rats.展开更多
The CRISPR-Cas13 systems have opened a new avenue for RNA detection due to their exceptional programmable collateral activity against ssRNA.However,most high-performance Cas13-based RNA sensors still suffer from some ...The CRISPR-Cas13 systems have opened a new avenue for RNA detection due to their exceptional programmable collateral activity against ssRNA.However,most high-performance Cas13-based RNA sensors still suffer from some drawbacks because of the tradeoffs between readout device complexity and analytical sensitivity,which can limit their viability at the point-of-care(POC).To overcome these shortcomings,we herein report a novel target-responsive POC platform for translating RNA detection into a glucose test.Specifically,this platform combines the advantages of three techniques:a hybrid Cas13a/Cas12a system for both RNA recognition with single-base resolution and cascade enzymatic amplification,a self-sustainable catalytic nucleic acid circuit(SCC)with embedded uracil-base and poly-T bulges for Cas13a-and Cas12amediated collateral cleavage,respectively,and a portable glucose meter(PGM)for simple signal readout.The incorporation of an engineered ssDNA–invertase conjugate in SCC led to RNA-to-glucose signal transduction through the hydrolysis of sucrose to glucose.By targeting a 20-nt conserved region of BMAL1 mRNA(mBMAL1),a key circadian clock gene,we demonstrate that SCC can serve as an excellent CRISPR reporter,enabling the direct detection of unamplified mBMAL1 as low as 0.864 fM concurrently with good discrimination ability for single-base mismatch.More importantly,this integrated platform was successfully applied for POC diagnosis of mBMAL1 from different cell lysates,with results that strongly correlate with RT-PCR.Given its wide adaptability,we anticipate that such a CRISPR-SCC system can be easily modified to quantify other RNA biomarkers associated with circadian rhythm.展开更多
The oriental armyworm,Mythimna separata,is a major long-distance migratory insect pest of grain crops in China and other Asian countries.Migratory flights and reproductive behavior usually occur at night,regulated by ...The oriental armyworm,Mythimna separata,is a major long-distance migratory insect pest of grain crops in China and other Asian countries.Migratory flights and reproductive behavior usually occur at night,regulated by a circadian rhythm.However,knowledge about the linkages between adult flight,reproduction,and clock genes is still incomplete.To fill this important gap in our knowledge,a clock gene(designated Msper)was identified and phylogenetic analysis indicated that the encoded protein(MsPER)was highly similar to PER proteins from other insect species.Quantitative RT-PCR assays demonstrated that significantly different spatiotemporal and circadian rhythmic accumulations of mRNA encoding MsPER occurred during development under steady 14 h:10 h light:dark conditions.The highest mRNA accumulation occurred in adult antennae and the lowest in larvae.Msper was expressed rhythmically in adult antennae,relatively less in photophase and more entering scotophase.Injecting small interference RNA(siRNA)into adult heads effectively knocked down Msper mRNA levels within 72 h.Most siRNA-injected adults reduced their evening flight activity significantly and did not exhibit a normal evening peak of flight activity.They also failed to mate and lay eggs within 72 h.Adult mating behavior was restored to control levels by 72 h post injection.We infer that Msper is a prominent clock gene that acts in regulating adult migratory flight and mating behaviors of M.separata.Because of its influence on migration and mating,Msper may be a valuable gene to target for effective management of this migratory insect.展开更多
Objective To understand the effects of clock gene BMAL1 and HIF-1α(Hypoxia inducible factor-1α)on proliferation,migration and sensitivity to radiotherapy of nasopharyngeal carcinoma cells HONE1.At the same time,whet...Objective To understand the effects of clock gene BMAL1 and HIF-1α(Hypoxia inducible factor-1α)on proliferation,migration and sensitivity to radiotherapy of nasopharyngeal carcinoma cells HONE1.At the same time,whether the biological clock gene BMAL1 can affect the expression of HIF-1αprotein was investigated.It will lay the foundation for further study on the correlation between clock gene BMAL1 and HIF pathway.Methods BMAL1 gene overexpression and interference lentivirus and HIF-1αgene interference lentivirus were constructed respectively,and were transfected into nasopharyngeal carcinoma cells HONE1.Western blot was used to verify the establishment of overexpressed and knockdown BMAL1 cell lines and HIF-1αgene knockdown cell line,and to investigate the expression of HIF-1αprotein in overexpressed and knockdown BMAL1 cell lines.CCK-8 cell proliferation test and scratch test were used to analyze the proliferation and migration ability of cells.Cell apoptosis after radiotherapy was analyzed by flow cytometry.The effects of BMAL1 and HIF-1αon the sensitivity of HONE1 radiotherapy in nasopharyngeal carcinoma cells after X-ray irradiation at different doses(0Gy,2Gy,4Gy,6Gy)were detected by clone formation assay.Results The overexpression of BMAL1 gene and lentivirus interference were constructed to effectively up regulate and down regulate the expression of BMAL1 protein in nasopharyngeal carcinoma cells HONE1.Meanwhile,HIF-1αgene interference lentivirus was constructed to effectively down-regulate the expression of HIF-1αprotein in nasopharyngeal carcinoma cell line HONE1,and successfully screen out stable nasopharyngeal carcinoma cell lines.Western blot results showed that overexpression of BMAL1 gene could inhibit the expression of HIF-1αprotein in HONE1 of nasopharyngeal carcinoma cells,while knockdown of BMAL1 gene promoted the expression of HIF-1αprotein in HONE1 of nasopharyngeal carcinoma cells(P<0.05).CCK-8 cell proliferation and scratch test showed that overexpression of BMAL1 gene or knockdown of HIF-1αgene could inhibit the proliferation and migration of HONE1 cells(P<0.05).Flow cytometry results showed that after 8Gy irradiation for 72 h,the apoptosis rate of BMALl gene overexpression group was higher than that of the overexpression control group,similarly,the apoptosis rate of HIF-1αgene knockdown group was higher than that of the knockdown control group(P<0.05).After X-ray irradiation at different doses(0Gy,2Gy,4Gy,6Gy),clon-formation experiment showed that the clon-formation rate and cell survival fraction of BMALl overexpression group or HIF-1αknockdown group were lower than those of negative control group(P<0.05).Sigmaplot analysis showed that the D0,Dq and SF2 of the BMAL1 overexpression group or HIF-1αknockdown group were lower than those of the negative control group,and the radiosensitization ratios were 1.381 and 1.063,respectively.Conclusion Overexpression of BMAL1 gene can inhibit the proliferation and migration of nasopharyngeal carcinoma cell line HONE1,increase apoptosis after radiotherapy and improve radiosensitivity.Knock down HIF-1αGene can inhibit the proliferation and migration of nasopharyngeal carcinoma cell line HONE1,increase apoptosis after radiotherapy and improve radiosensitivity.In nasopharyngeal carcinoma cells HONE1,overexpression of BMAL1 gene can inhibit the expression of HIF-1αprotein while knockdown of BMAL1 gene can promote the expression of HIF-1αprotein.展开更多
Circadian clocks are endogenous timers that enable plants to synchronize biological processes with daily and seasonal environmental conditions in order to allocate resources during the most beneficial times of day and...Circadian clocks are endogenous timers that enable plants to synchronize biological processes with daily and seasonal environmental conditions in order to allocate resources during the most beneficial times of day and year. The circadian clock regulates a number of central plant activities, including growth, develop- ment, and reproduction, primarily through controlling a substantial proportion of transcriptional activity and protein function. This review examines the roles that alleles of circadian clock genes have played in domestication and improvement of crop plants. The focus here is on three groups of circadian clock genes essential to clock function in Arabidopsis thaliana： PSEUDO-RESPONSE REGULATORs, GIGANTEA, and the evening complex genes EARL Y FLOWERING 3, EARLY FLOWERING 4, and LUX ARRHYTHMO. Homol- ogous genes from each group underlie quantitative trait loci that have beneficial influences on key agricul- tural traits, especially flowering time but also yield, biomass, and biennial growth habit. Emerging insights into circadian clock regulation of other fundamental plant processes, including responses to abiotic and biotic stresses, are discussed to highlight promising avenues for further crop improvement.展开更多
In insects,facultative diapause is a state of developmental arrest mainly induced by photoperiod or temperature that allows insects to survive adverse environmental conditions.Understanding how insect initiates facult...In insects,facultative diapause is a state of developmental arrest mainly induced by photoperiod or temperature that allows insects to survive adverse environmental conditions.Understanding how insect initiates facultative diapause and prepares diapause can provide us new insights to study developmental and evolutionary biology.It has been shown that the circadian clock genes can participate in photoperiodic measurement and regulate reproductive diapause initiation through JH signaling in short-day-induced winter diapause.However,how circadian clock genes translate photoperiodic information into downstream JH signaling for diapause destiny and then affect diapause preparation remains largely unknown.In the present study,we investigate this in the cabbage beetle Colaphellus bowringi which undergoes reproductive diapause under long-day condition.We respectively knocked down two circadian clock negative regulators,period(per)and timeless(tim),in the 3-day-old larvae(most sensitive to photoperiod),and dsgfp treatment was served as a control.Under the diapause-inducing photoperiod(16L:8D),knocking down per and tim significantly decreased the rate of burrowing behavior.And mtany female beetles of the per and tim RNAi showed developed ovary,decreased lipid accumulation and downregulated expression of stress resistance genes.The JHinduced genes,Kr-h1,JHE1,Vg1,and Vg2, significantly increased in the females with suppression of per and tim.It implied that suppression of per and tim during diapause initiation phase(DIP)could activate the JH signaling in the female adults.Before the beetles enter into diapause preparation phase(DPP),we used RNA sequencing to analyize gene expression profiles after per and tim RNAi.It showed that many differentially expressed genes were enriched in environmental information processing,such as mTOR and TGF-beta signaling pathway.To ask whether per and tim also regulate diapause preparation,we knocked down these two genes in the female adults during DPP.It showed that the diapause destiny was not af-fected,but the lipid storage in diapause-destined females was significantly reduced after per and tim RNAi.Interestingly,per-and tim-regulated lipid storage during DPP was independent on JH signaling.We further found that per and tim promoted lipid storage by regulating the expression of genes that control lipogenesis and lipolysis.In summary,these results suggest that per and tim participate in photoperiodic measurement and initiate reproductive diapause through JH signaling during DIP in long-day-induced summer diapause.The mTOR and TGF-beta signaling pathways may be involved in the regulation of JH signaling by circadian clock genes.Meanwhile,per and tim transduce photoperiodic signal and promote lipid storage during DPP in a JH-independent manner.These results provide us new clues to study the molecular mechanism of photoperiod-regulated diapause induction in insects.展开更多
文摘Objective To investigate the structural changes of rat thoracic aorta and changes in expression levels of Bmal1 and cyclins in thoracic aorta endothelial cells following heat stress.Methods Twenty male SD rats were randomized equally into control group and heat stress group.After exposure to 32℃for 2 weeks in the latter group,the rats were examined for histopathological changes and Bmal1 expression in the thoracic aorta using HE staining and immunohistochemistry.In the cell experiments,cultured rat thoracic aortic endothelial cells(RTAECs)were incubated at 40℃for 12 h with or without prior transfection with a Bmal1-specific small interfering RNA(si-Bmal1)or a negative sequence.In both rat thoracic aorta and RTAECs,the expressions of Bmal1,the cell cycle proteins CDK1,CDK4,CDK6,and cyclin B1,and apoptosis-related proteins Bax and Bcl-2 were detected using Western blotting.TUNEL staining was used to detect cell apoptosis in rat thoracic aorta,and the changes in cell cycle distribution and apoptosis in RTAECs were analyzed with flow cytometry.Results Compared with the control rats,the rats exposed to heat stress showed significantly increased blood pressures and lowered heart rate with elastic fiber disruption and increased expressions of Bmal1,cyclin B1 and CDK1 in the thoracic aorta(P<0.05).In cultured RTAECs,heat stress caused significant increase of Bmal1,cyclin B1 and CDK1 protein expression levels,which were obviously lowered in cells with prior si-Bmal1 transfection.Bmal1 knockdown also inhibited heat stress-induced increase of apoptosis in RTAECs as evidenced by decreased expression of Bax and increased expression of Bcl-2.Conclusion Heat stress upregulates Bmal1 expression and causes alterations in expressions of cyclins to trigger apoptosis of rat thoracic aorta endothelial cells,which can be partly alleviated by suppressing Bmal1 expression.
基金supported by grants from the National Natural Science Foundation of China(N0.82474525 and No.82074444)the Hunan Provincial Natural Outstanding Young People Science Foundation(2023JJ10032)the Hunan Province Health and High-Level Talent Medical Academic Leader Training Plan(20240304051).
文摘The circadian clock is an important internal time regulatory system for a range of physiological and behavioral rhythms within living organisms.Testosterone,as one of the most critical sex hormones,is essential for the development of the reproductive system,maintenance of reproductive function,and the overall health of males.The secretion of testosterone in mammals is characterized by distinct circadian rhythms and is closely associated with the regulation of circadian clock genes.Here we review the central and peripheral regulatory mechanisms underlying the influence of circadian clock genes upon testosterone synthesis.We also examined the specific effects of these genes on the occurrence,development,and treatment of common male diseases,including late-onset hypogonadism,erectile dysfunction,male infertility,and prostate cancer.
文摘Clock genes create a complicated molecular time-keeping system consisting of multiple positive and negative feedback loops at transcriptional and translational levels. This circadian system coordinates and regulates multiple cellular procedures implicated in cancer development such as metabolism, cell cycle and DNA damage response. Recent data support that molecules such as CLOCK1, BMAL1 and PER and CRY proteins have various effects on c-Myc/p21 and Wnt/β-catenin pathways and influence multiple steps of DNA damage response playing a critical role in the preservation of genomic integrity in normal and cancer cells. Notably, all these events have already been related to the development and progression of colorectal cancer (CRC). Recent data highlight critical correlations between clock genes’ expression and pathogenesis, progression, aggressiveness and prognosis of CRC. Increased expression of positive regulators of this circadian system such as BMAL1 has been related to decrease overall survival while decreased expression of negative regulators such as PER2 and PER3 is connected with poorer differentiation, increased aggressiveness and worse prognosis. The implications of these molecules in DNA repair systems explain their involvement in the development of CRC but at the same time provide us with novel targets for modern therapeutic approaches for patients with advanced CRC.
基金This work was supported by the National Natural Science Foundation of China(Nos.81901430 and 81871835)the Guangdong Provincial Natural Science Foundation of China(No.2022A1515010379)+1 种基金the Innovation Project from Department of Education of Guangdong Province(No.2021KTSCX 055)the Shanghai Frontiers Science Research Base of Exercise and Metabolic Health,and the Shanghai Key Laboratory for Human Athletic Ability Development and Support(Shanghai University of Sport)(No.11DZ2261100),China.
文摘Mammalian bone is constantly metabolized from the embryonic stage,and the maintenance of bone health depends on the dynamic balance between bone resorption and bone formation,mediated by osteoclasts and osteoblasts.It is widely recognized that circadian clock genes can regulate bone metabolism.In recent years,the regulation of bone metabolism by non-coding RNAs has become a hotspot of research.MicroRNAs can participate in bone catabolism and anabolism by targeting key factors related to bone metabolism,including circadian clock genes.However,research in this field has been conducted only in recent years and the mechanisms involved are not yet well established.Recent studies have focused on how to target circadian clock genes to treat some diseases,such as autoimmune diseases,but few have focused on the co-regulation of circadian clock genes and microRNAs in bone metabolic diseases.Therefore,in this paper we review the progress of research on the co-regulation of bone metabolism by circadian clock genes and microRNAs,aiming to provide new ideas for the prevention and treatment of bone metabolic diseases such as osteoporosis.
基金Supported by the National Natural Science Foundation of China(No.31402305)the Educational Commission of Sichuan Province of China(No.14ZA0249)+1 种基金the Key Technologies R&D Program of Neijiang,Sichuan,China(No.12108)the College Students’ Scientific Research Project of Neijiang Normal University(Nos.13NSD-66,13NSD-77)
文摘The Clock gene,a key molecule in circadian systems,is widely distributed in the animal kingdom. We isolated a 936-bp partial c DNA sequence of the C lock gene( Pva- clock) from the darkbarbel catfish P elteobagrus vachelli that exhibited high identity with C lock genes of other species of fish and animals(65%–88%). The putative domains included a basic helix-loop-helix(b HLH) domain and two period-ARNT-single-minded(PAS) domains,which were also similar to those in other species of fish and animals. P va- Clock was primarily expressed in the brain,and was detected in all of the peripheral tissues sampled. Additionally,the pattern of P va- Clock expression over a 24-h period exhibited a circadian rhythm in the brain,liver and intestine,with the acrophase at zeitgeber time 21:35,23:00,and 23:23,respectively. Our results provide insight into the function of the molecular C lock of P. vachelli.
基金supported by grants from the Department of Public Health of Hubei Province of China (No. 2012Z-B08)the Health Bureau of Wuhan City of China (No. WX12C10)
文摘Summary： This study investigated the effects of benazepril administered in the morning or evening on the diurnal variation of renin-angiotensin-aldosterone system （RAAS） and clock genes in the kidney. The male Wistar rat models of 5/6 subtotal nephrectomy （STNx） were established. Animals were ran- domly divided into 4 groups： sham STNx group （control）, STNx group, morning benazepril group （MB） and evening benazepril group （EB）. Benazepril was intragastfically administered at a dose of 10 mg/kg/day at 07：00 and 19：00 in the MB group and EB group respectively for 12 weeks. All the animals were synchronized to the light：dark cycle of 12：12 for 12 weeks. Systolic blood pressure （SBP）, 24-h urinary protein excretion and renal function were measured at 11 weeks. Blood samples and kidneys were collected every 4 h throughout a day to detect the expression pattern of renin activity （RA）, angio- tensin Ⅱ （Ang Ⅱ ） and aldosterone （Aid） by radioimmunoassay （RIA） and the mRNA expression profile of clock genes （bmall, dbp and per2） by real-time PCR at 12 weeks. Our results showed that no signifi- cant differences were noted in the SBP, 24-h urine protein excretion and renal function between the MB and EB groups. There were no significant differences in average Aid and RA content of a day between the MB group and EB group. The expression peak of bmall mRNA was phase-delayed by 4 to 8 h, and the diurnal variation of per2 and dbp mRNA diminished in the MB and EB groups compared with the control and STNx groups. It was concluded when the similar SBP reduction, RAAS inhibition and clock gene profile were achieved with optimal dose of benazepril, morning versus evening dosing of benazepril has the same renoprotection effects.
基金supported by the Program for Guangdong Introducing Innovative and Entrepreneurial Teams(2019ZT08N291)the Science and Technology Program of Guangzhou,China(2023A04J0760)the Guangdong Basic and Applied Basic Research Foundation(2024A1515030058).
文摘As the body’s internal clock,the circadian rhythm regulates the energy expenditure,appetite,and sleep.There exists a close relationship between the host circadian rhythm and gut microbiota.In this work,a circadian disorder mouse model induced by constant darkness(CD)was constructed to investigate the regulating effects of capsaicin(CAP)on disturbances of metabolism homeostasis and gut microbiota in the respect of circadian rhythm-related mechanisms.Our results indicated that CAP reduced weight gain induced by circadian rhythm disorder in mice by inhibiting fat accumulation in liver and adipose tissue.The rhythmic expressions of circadian clock genes and lipid-metabolism related genes in liver were also recovered by CAP.Microbial study using 16S rRNA sequencing revealed that CAP modulated the gut microbiota richness,diversity and composition,and restored diurnal oscillations of gut microbes at the phylum and family level.These results indicated that CAP could alleviate CD-induced hepatic clock gene disruption and gut microbiota dysbiosis in mice,providing theoretical basis for CAP to be used as a muti-functional ingredient with great healthpromoting effects.
文摘Objective： Investigation of the effect of Xiaoaiping on the expression of circadian clock genes in human hepatoma HepG2 cells. Methods： Selecting the HepG2 cells in the logarithmic growth phase and assigning them to Xiaoaiping injection （XAP） group and control group. The two groups were treated with 75 mg/mL XAP or the same dose of normal saline. After 72 h of treatment, real-time PCR was used to detect the expression of circadian clock genes in HepG2 cells and Western Blot technology was used to detect the expression of related proteins. Results： The mRNA expression levels of PER1, NPAS2, NR1D1, and DEC1 in the XAP group was significantly higher than that in the control group （P〈 0.05）, while the mRNA expression levels of PER3, BMAL1, DEC2, and RORA were significantly lower in the XAP group than in the control group （P 〈 0.05）, and there was no significant difference between the mRNA expression levels of PER2, CRY1, CRY2, and TIM. Of course, the proteins＇ expression levels of the genes we had detected such as PERle3, CRYI-2, CLOCK, BMAL1 by Western Blot were consistent with the real-time PCR results above. Conclusion： XAP affects the expression of circadian clock genes in HepG2 cells.
基金Supported by RC1303GA49 and Italian Ministry of Health(Pazienza V)MV and VP are supported by Bando GR-2010-2311017 and by the"5x1000"voluntary contributions to IRCCS"Casa Sollievo della Sofferenza"Hospital(Vinciguerra M and Pazienza V)and the Associazione Italiana per la Ricerca sul Cancro(AIRC)program MyFAG(Vinciguerra M)
文摘Many aspects of cellular physiology display circadian(approximately 24-h)rhythms.Dysfunction of the circadian clock molecular circuitry is associated with human health derangements,including neurodegeneration,increased risk of cancer,cardiovascular diseases and the metabolic syndrome.Viruses triggering hepatitis depend tightly on the host cell synthesis machinery for their own replication,survival and spreading.Recent evidences support a link between the circadian clock circuitry and viruses’biological cycle within host cells.Currently,in vitro models for chronobiological studies of cells infected with viruses need to be implemented.The establishment of such in vitro models would be helpful to better understand the link between the clock gene machinery and viral replication/viral persistence in order to develop specifically targeted therapeutic regimens.Here we review the recent literature dealing with the interplay between hepatitis B and C viruses and clock genes.
文摘Objective: To investigate the effects of 72 hours continuous sleep deprivation (SD) on circadian clock gene expression and oxidative stress in the rat liver. Methods: Twenty healthy male Sprague-Dawley rats were divided into 2 groups (n = 10 each) using a random number table: normal control group (group C), sleep deprivation group (group SD). Group SD was treated with a modified multiple platform water environment method. After 72 hours sleep deprived, the levels of AST (Aspartate transaminase ) and ALT (Alanine aminotransferase) in serum were determined. The contents of malondialdehyde (MDA), the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the liver tissue of the rats were examined in both two groups. The expression levels of CLOCK, BMAL1 and CRY1 protein in liver tissue were examined by Western blotting. Results: Compared with group C, the content of MDA, and the levels of AST and ALT in serum were significantly increased (P Conclusion: 72 hours continuous sleep deprivation can downregulate the expression of circadian clock gene and promote oxidative stress in rats.
基金supported by the National Natural Science Foundation of China(No.22274072)。
文摘The CRISPR-Cas13 systems have opened a new avenue for RNA detection due to their exceptional programmable collateral activity against ssRNA.However,most high-performance Cas13-based RNA sensors still suffer from some drawbacks because of the tradeoffs between readout device complexity and analytical sensitivity,which can limit their viability at the point-of-care(POC).To overcome these shortcomings,we herein report a novel target-responsive POC platform for translating RNA detection into a glucose test.Specifically,this platform combines the advantages of three techniques:a hybrid Cas13a/Cas12a system for both RNA recognition with single-base resolution and cascade enzymatic amplification,a self-sustainable catalytic nucleic acid circuit(SCC)with embedded uracil-base and poly-T bulges for Cas13a-and Cas12amediated collateral cleavage,respectively,and a portable glucose meter(PGM)for simple signal readout.The incorporation of an engineered ssDNA–invertase conjugate in SCC led to RNA-to-glucose signal transduction through the hydrolysis of sucrose to glucose.By targeting a 20-nt conserved region of BMAL1 mRNA(mBMAL1),a key circadian clock gene,we demonstrate that SCC can serve as an excellent CRISPR reporter,enabling the direct detection of unamplified mBMAL1 as low as 0.864 fM concurrently with good discrimination ability for single-base mismatch.More importantly,this integrated platform was successfully applied for POC diagnosis of mBMAL1 from different cell lysates,with results that strongly correlate with RT-PCR.Given its wide adaptability,we anticipate that such a CRISPR-SCC system can be easily modified to quantify other RNA biomarkers associated with circadian rhythm.
基金the National Natural Science Foundation of China(No.32072420,31871951,31672019)China Agriculture Research System of MOF and MARA(CARS-22)the National Key Research and Development Program of China(2017YFD0201802,2017YFD0201701).
文摘The oriental armyworm,Mythimna separata,is a major long-distance migratory insect pest of grain crops in China and other Asian countries.Migratory flights and reproductive behavior usually occur at night,regulated by a circadian rhythm.However,knowledge about the linkages between adult flight,reproduction,and clock genes is still incomplete.To fill this important gap in our knowledge,a clock gene(designated Msper)was identified and phylogenetic analysis indicated that the encoded protein(MsPER)was highly similar to PER proteins from other insect species.Quantitative RT-PCR assays demonstrated that significantly different spatiotemporal and circadian rhythmic accumulations of mRNA encoding MsPER occurred during development under steady 14 h:10 h light:dark conditions.The highest mRNA accumulation occurred in adult antennae and the lowest in larvae.Msper was expressed rhythmically in adult antennae,relatively less in photophase and more entering scotophase.Injecting small interference RNA(siRNA)into adult heads effectively knocked down Msper mRNA levels within 72 h.Most siRNA-injected adults reduced their evening flight activity significantly and did not exhibit a normal evening peak of flight activity.They also failed to mate and lay eggs within 72 h.Adult mating behavior was restored to control levels by 72 h post injection.We infer that Msper is a prominent clock gene that acts in regulating adult migratory flight and mating behaviors of M.separata.Because of its influence on migration and mating,Msper may be a valuable gene to target for effective management of this migratory insect.
基金supported in part by grants from the National Natural Science Foundation of China under grant number 82060556,81560437the Department of Science and Technology,Guizhou Province,under grant number[2018]2755+3 种基金the Ordinary Colleges and Universities Youth Science and Technology Talent Growth Project,Guizhou Province,under grant number[2021]187The Health Commission Science and Technology Fund,Guizhou Provincial under grant number gzwkj2021-050Guizhou Medical University 2021 National Foundation Cultivation Project[20NSP041]the Hospital-level Science and Technology Project of Guizhou Cancer Hospital under grant number YJ2019-33.
文摘Objective To understand the effects of clock gene BMAL1 and HIF-1α(Hypoxia inducible factor-1α)on proliferation,migration and sensitivity to radiotherapy of nasopharyngeal carcinoma cells HONE1.At the same time,whether the biological clock gene BMAL1 can affect the expression of HIF-1αprotein was investigated.It will lay the foundation for further study on the correlation between clock gene BMAL1 and HIF pathway.Methods BMAL1 gene overexpression and interference lentivirus and HIF-1αgene interference lentivirus were constructed respectively,and were transfected into nasopharyngeal carcinoma cells HONE1.Western blot was used to verify the establishment of overexpressed and knockdown BMAL1 cell lines and HIF-1αgene knockdown cell line,and to investigate the expression of HIF-1αprotein in overexpressed and knockdown BMAL1 cell lines.CCK-8 cell proliferation test and scratch test were used to analyze the proliferation and migration ability of cells.Cell apoptosis after radiotherapy was analyzed by flow cytometry.The effects of BMAL1 and HIF-1αon the sensitivity of HONE1 radiotherapy in nasopharyngeal carcinoma cells after X-ray irradiation at different doses(0Gy,2Gy,4Gy,6Gy)were detected by clone formation assay.Results The overexpression of BMAL1 gene and lentivirus interference were constructed to effectively up regulate and down regulate the expression of BMAL1 protein in nasopharyngeal carcinoma cells HONE1.Meanwhile,HIF-1αgene interference lentivirus was constructed to effectively down-regulate the expression of HIF-1αprotein in nasopharyngeal carcinoma cell line HONE1,and successfully screen out stable nasopharyngeal carcinoma cell lines.Western blot results showed that overexpression of BMAL1 gene could inhibit the expression of HIF-1αprotein in HONE1 of nasopharyngeal carcinoma cells,while knockdown of BMAL1 gene promoted the expression of HIF-1αprotein in HONE1 of nasopharyngeal carcinoma cells(P<0.05).CCK-8 cell proliferation and scratch test showed that overexpression of BMAL1 gene or knockdown of HIF-1αgene could inhibit the proliferation and migration of HONE1 cells(P<0.05).Flow cytometry results showed that after 8Gy irradiation for 72 h,the apoptosis rate of BMALl gene overexpression group was higher than that of the overexpression control group,similarly,the apoptosis rate of HIF-1αgene knockdown group was higher than that of the knockdown control group(P<0.05).After X-ray irradiation at different doses(0Gy,2Gy,4Gy,6Gy),clon-formation experiment showed that the clon-formation rate and cell survival fraction of BMALl overexpression group or HIF-1αknockdown group were lower than those of negative control group(P<0.05).Sigmaplot analysis showed that the D0,Dq and SF2 of the BMAL1 overexpression group or HIF-1αknockdown group were lower than those of the negative control group,and the radiosensitization ratios were 1.381 and 1.063,respectively.Conclusion Overexpression of BMAL1 gene can inhibit the proliferation and migration of nasopharyngeal carcinoma cell line HONE1,increase apoptosis after radiotherapy and improve radiosensitivity.Knock down HIF-1αGene can inhibit the proliferation and migration of nasopharyngeal carcinoma cell line HONE1,increase apoptosis after radiotherapy and improve radiosensitivity.In nasopharyngeal carcinoma cells HONE1,overexpression of BMAL1 gene can inhibit the expression of HIF-1αprotein while knockdown of BMAL1 gene can promote the expression of HIF-1αprotein.
文摘Circadian clocks are endogenous timers that enable plants to synchronize biological processes with daily and seasonal environmental conditions in order to allocate resources during the most beneficial times of day and year. The circadian clock regulates a number of central plant activities, including growth, develop- ment, and reproduction, primarily through controlling a substantial proportion of transcriptional activity and protein function. This review examines the roles that alleles of circadian clock genes have played in domestication and improvement of crop plants. The focus here is on three groups of circadian clock genes essential to clock function in Arabidopsis thaliana： PSEUDO-RESPONSE REGULATORs, GIGANTEA, and the evening complex genes EARL Y FLOWERING 3, EARLY FLOWERING 4, and LUX ARRHYTHMO. Homol- ogous genes from each group underlie quantitative trait loci that have beneficial influences on key agricul- tural traits, especially flowering time but also yield, biomass, and biennial growth habit. Emerging insights into circadian clock regulation of other fundamental plant processes, including responses to abiotic and biotic stresses, are discussed to highlight promising avenues for further crop improvement.
基金Funded by the National Natural Science Foundation of China(31701842)
文摘In insects,facultative diapause is a state of developmental arrest mainly induced by photoperiod or temperature that allows insects to survive adverse environmental conditions.Understanding how insect initiates facultative diapause and prepares diapause can provide us new insights to study developmental and evolutionary biology.It has been shown that the circadian clock genes can participate in photoperiodic measurement and regulate reproductive diapause initiation through JH signaling in short-day-induced winter diapause.However,how circadian clock genes translate photoperiodic information into downstream JH signaling for diapause destiny and then affect diapause preparation remains largely unknown.In the present study,we investigate this in the cabbage beetle Colaphellus bowringi which undergoes reproductive diapause under long-day condition.We respectively knocked down two circadian clock negative regulators,period(per)and timeless(tim),in the 3-day-old larvae(most sensitive to photoperiod),and dsgfp treatment was served as a control.Under the diapause-inducing photoperiod(16L:8D),knocking down per and tim significantly decreased the rate of burrowing behavior.And mtany female beetles of the per and tim RNAi showed developed ovary,decreased lipid accumulation and downregulated expression of stress resistance genes.The JHinduced genes,Kr-h1,JHE1,Vg1,and Vg2, significantly increased in the females with suppression of per and tim.It implied that suppression of per and tim during diapause initiation phase(DIP)could activate the JH signaling in the female adults.Before the beetles enter into diapause preparation phase(DPP),we used RNA sequencing to analyize gene expression profiles after per and tim RNAi.It showed that many differentially expressed genes were enriched in environmental information processing,such as mTOR and TGF-beta signaling pathway.To ask whether per and tim also regulate diapause preparation,we knocked down these two genes in the female adults during DPP.It showed that the diapause destiny was not af-fected,but the lipid storage in diapause-destined females was significantly reduced after per and tim RNAi.Interestingly,per-and tim-regulated lipid storage during DPP was independent on JH signaling.We further found that per and tim promoted lipid storage by regulating the expression of genes that control lipogenesis and lipolysis.In summary,these results suggest that per and tim participate in photoperiodic measurement and initiate reproductive diapause through JH signaling during DIP in long-day-induced summer diapause.The mTOR and TGF-beta signaling pathways may be involved in the regulation of JH signaling by circadian clock genes.Meanwhile,per and tim transduce photoperiodic signal and promote lipid storage during DPP in a JH-independent manner.These results provide us new clues to study the molecular mechanism of photoperiod-regulated diapause induction in insects.
基金This work was supported by the National Natural Science Foundation of China(No.30170295),Medical Developmental Foundation of Soochow University(No.EEl 34031) and Young Teacher's Research Foundation of Soochow University(No.Q3134044).
