Diabetic nephropathy is one of the various complications of diabetes mellitus, affecting patients for lifetime. Earlier studies have revealed that genipin can not only improve diabetes, but also induce cytotoxicity. T...Diabetic nephropathy is one of the various complications of diabetes mellitus, affecting patients for lifetime. Earlier studies have revealed that genipin can not only improve diabetes, but also induce cytotoxicity. Therefore, it is not clear which effect of genipin on kidneys occurs, when it is used in the treatment of diabetes. In the present study, we performed nuclear magnetic resonance(NMR)-based metabolomics analysis of urine and kidney tissue samples obtained from diabetic rats to explore the change of endogenous metabolites associated with diabetes and concomitant kidney disease. Nine significant differential metabolites that were closely related to renal function were screened. They were mainly related to three metabolic pathways: synthesis and degradation of ketone bodies, glycine, serine and threonine metabolism, and butanoate metabolism, which are involved in methylamine metabolism, energy metabolism and amino acid metabolism. In addition, after the intervention of genipin, the metabolic levels of all the metabolites tended to be normal, indicating a protective effect of genipin on kidneys. Our results may be helpful for understanding the antidiabetic effect of genipin.展开更多
BACKGROUND Glycation is an important step in aging and oxidative stress,which can lead to endothelial dysfunction and cause severe damage to the eyes or kidneys of diabetics.Inhibition of the formation of advanced gly...BACKGROUND Glycation is an important step in aging and oxidative stress,which can lead to endothelial dysfunction and cause severe damage to the eyes or kidneys of diabetics.Inhibition of the formation of advanced glycation end products(AGEs)and their cell toxicity can be a useful therapeutic strategy in the prevention of diabetic retinopathy(DR).Gardenia jasminoides Ellis(GJE)fruit is a selective inhibitor of AGEs.Genipin is an active compound of GJE fruit,which can be employed to treat diabetes.AIM To confirm the effect of genipin,a vital component of GJE fruit,in preventing human retinal microvascular endothelial cells(hRMECs)from AGEs damage in DR,to investigate the effect of genipin in the down-regulation of AGEs expression,and to explore the role of the CHGA/UCP2/glucose transporter 1(GLUT1)signal pathway in this process.METHODS In vitro,cell viability was tested to determine the effects of different doses of glucose and genipin in hRMECs.Cell Counting Kit-8(CCK-8),colony formation assay,flow cytometry,immunofluorescence,wound healing assay,transwell assay,and tube-forming assay were used to detect the effect of genipin on hRMECs cultured in high glucose conditions.In vivo,streptozotocin(STZ)induced mice were used,and genipin was administered by intraocular injection(IOI).To explore the effect and mechanism of genipin in diabetic-induced retinal dysfunction,reactive oxygen species(ROS),mitochondrial membrane potential(MMP),and 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose(2-NBDG)assays were performed to explore energy metabolism and oxidative stress damage in high glucose-induced hRMECs and STZ mouse retinas.Immunofluorescence and Western blot were used to investigate the expression of inflammatory cytokines[vascular endothelial growth factor(VEGF),SCG3,tumor necrosis factor-alpha(TNF-α),interleukin(IL)-1β,IL-18,and nucleotide-binding domain,leucine-rich-containing family,pyrin domain-containing 3(NLRP3)].The protein expression of the receptor of AGEs(RAGE)and the mitochondria-related signal molecules CHGA,GLUT1,and UCP2 in high glucose-induced hRMECs and STZ mouse retinas were measured and compared with the genipin-treated group.RESULTS The results of CCK-8 and colony formation assay showed that genipin promoted cell viability in high glucose(30 mmol/L D-Glucose)-induced hRMECs,especially at a 0.4μmol/L dose for 7 d.Flow cytometry results showed that high glucose can increase apoptosis rate by 30%,and genipin alleviated cell apoptosis in AGEs-induced hRMECs.A high glucose environment promoted ATP,ROS,MMP,and 2-NBDG levels,while genipin inhibited these phenotypic abnormalities in AGEs-induced hRMECs.Furthermore,genipin remarkably reduced the levels of the pro-inflammatory cytokines TNF-α,IL-1β,IL-18,and NLRP3 and impeded the expression of VEGF and SCG3 in AGEs-damaged hRMECs.These results showed that genipin can reverse high glucose induced damage with regard to cell proliferation and apoptosis in vitro,while reducing energy metabolism,oxidative stress,and inflammatory injury caused by high glucose.In addition,ROS levels and glucose uptake levels were higher in the retina from the untreated eye than in the genipin-treated eye of STZ mice.The expression of inflammatory cytokines and pathway protein in the untreated eye compared with the genipin-treated eye was significantly increased,as measured by Western blot.These results showed that IOI of genipin reduced the expression of CHGA,UCP2,and GLUT1,maintained the retinal structure,and decreased ROS,glucose uptake,and inflammation levels in vivo.In addition,we found that SCG3 expression might have a higher sensitivity in DR than VEGF as a diagnostic marker at the protein level.CONCLUSION Our study suggested that genipin ameliorates AGEs-induced hRMECs proliferation,apoptosis,energy metabolism,oxidative stress,and inflammatory injury,partially via the CHGA/UCP2/GLUT1 pathway.Control of advanced glycation by IOI of genipin may represent a strategy to prevent severe retinopathy and vision loss.展开更多
·AIM: To study the feasibility of using the discoloration to evaluate the biomechanical properties after treating with genipin.·METHODS: Porcine cadaver eyes were treated for30 min with 1.0%(by w/v) genipin....·AIM: To study the feasibility of using the discoloration to evaluate the biomechanical properties after treating with genipin.·METHODS: Porcine cadaver eyes were treated for30 min with 1.0%(by w/v) genipin. Untreated samples were used as controls. After treatment, scleral strips of4.0 ×10.0-mm2 were cut. The denaturation temperature(Td) measurement and stress-strain test were performed after taking photograph to analyze the color.·RESULTS: Within 24 h after treating with genipin, the sclera exhibited a bluish color which became deeper with time. And the denaturation temperature also was increased gradually. Compared with untreated groups, at1, 6, 12, 24 and 36 h after treatment, the ultimate stress were increased by 56%, 153%, 173%, 225% and 211%respectively. The Young’s modulus at 10% strain also increased by 170%, 246%, 264%, 389% and 288%respectively. There were strong correlation between the discoloration and the biomechanical properties(ΔE-Ultimate stress:R2=0.892, P =0.00; ΔE-Young’s modulus:R2=0.602, P =0.00).·CONCLUSION: Genipin could be used to strengthen collagen gradually in a relatively short time span. And the biomechanical properties could be reliably evaluated via simple visible discoloration.展开更多
In the present study,we aimed to ascertain the metabolic detoxification routes of genipin via CYP3A4,SULT2 A1,and UGT1 A1 in Hepa RG cells.It was found that the hepatic CYP3A4,SULT2 A1,and UGT1 A1 synergistically medi...In the present study,we aimed to ascertain the metabolic detoxification routes of genipin via CYP3A4,SULT2 A1,and UGT1 A1 in Hepa RG cells.It was found that the hepatic CYP3A4,SULT2 A1,and UGT1 A1 synergistically mediated the metabolic detoxification of genipin,and the CYP3A4 was the limited enzyme.In detail,the pivotal detoxification pathway was CYP3A4-SULT2 A1/UGT1 A1,indicating that SULT2 A1 and UGT1 A1 further catalyzed the phase II detoxification metabolism followed by the genipin metabolization by CYP3A4 to the phase I metabolites with alleviated toxicity.Our findings provided valuable cues for future studies on the detoxification of genipin,even the compatibility detoxification of Zhi-zi.Moreover,these data facilitated the development and rational administration of genipin and Zhi-zi.展开更多
AIM：To strengthen the biomechanics of collagen by crosslinking rabbit scleral collagen with genipin to develop a new therapy for preventing myopic progression. METHODS：Ten New Zealand rabbits were treated with 0.5 m...AIM：To strengthen the biomechanics of collagen by crosslinking rabbit scleral collagen with genipin to develop a new therapy for preventing myopic progression. METHODS：Ten New Zealand rabbits were treated with 0.5 mmol/L genipin injected into the sub-Tenon＇s capsule in the right eyes. Untreated contralateral eyes served as the control. The treated area was cut into scleral strips measuring 4.0 mm×10.0 mm for stress-strain measurements（n=5）. The remaining five treated eyes were prepared for histological examination.RESULTS：Compared to the untreated scleral strips,the genipin-crosslinked scleral strips showed that the ultimate stress and Young＇s modulus at 10% strain were increased by the amplitude of 130% and 303% respectively,ultimate strain was decreased by 24%. There had no α-smooth muscle actin（α-SMA）positive cells in control and treated sclera. Histologically,there was no sign of apoptosis in the sclera,choroid,and retina; and no side effects were found in the peripheral cornea and optic nerve adjacent to the treatment area.CONCLUSION：Genipin induced crosslinking of collagen can increase its biomechanical behavior by direct strengthening of the extracellular matrix in rabbit sclera,with no α-SMA expression seen in the myofibroblasts. As there is no evidence of cytotoxicity in the scleral,choroidal,and retinal cells,genipin is likely a promising agent to strengthen the weakened sclera to prevent myopic progression.展开更多
Antimicrobial delivery has been advocated for guided tissue regeneration (GTR) or guided bone regeneration (GBR) therapies involving patients with aggressive or unresolved periodontitis/peri-implantitis. Electrospun c...Antimicrobial delivery has been advocated for guided tissue regeneration (GTR) or guided bone regeneration (GBR) therapies involving patients with aggressive or unresolved periodontitis/peri-implantitis. Electrospun chitosan membranes demonstrate several advantages over traditional GTR barrier membranes because they stimulate healing, mimic the topology of the extracellular matrix, and allow for diffusion of nutrients and wastes into/out of the graft site, and were shown to stimulate bone formation in a rabbit calvarial criticalsize defect model. Previously, we have shown improvements in mechanical properties and degradation kinetics by crosslinking electrospun membranes with 5 mM or 10 mM genipin. We have also demonstrated the ability of elecrospun chitosan membranes to inhibit lippopolysaccharide (LPS)-induced monocyte activation. In this study, minocycline was incorporated into the chitosan membrane by passive absorption at 5 or 10 mg/mL. The minocycline-loaded membranes and control membranes (carrier only) were tested against Porphyromonas gingivalis (P. gingivalis) by repeated zone of inhibition (ZOI) measurements. Testing showed that uncrosslinked and genipin-crosslinked membranes have similar capacity to absorb aqueous solutions (swelling ratio 1.7 - 2.2). Minocycline loading resulted in bacterial inhibition for up to 8 days from crosslinked membranes (with 11 mm initial ZOI) whereas uncrosslinked membranes loaded with minocycline only inhibited bacteria for 4 days (with 8 mm initial ZOI). These in vitro results suggest that genipin-crosslinked electrospun chitosan membranes loaded with minocycline may be able to reduce early bacterial contamination of GTR graft sites.展开更多
Mitochondria are essential for eukaryotic life as powerhouses for energy metabolism. Excessive mitochondrial hyperthermia and reactive oxygen species(ROS) production have been associated with aging, cancer,neurodegene...Mitochondria are essential for eukaryotic life as powerhouses for energy metabolism. Excessive mitochondrial hyperthermia and reactive oxygen species(ROS) production have been associated with aging, cancer,neurodegenerative diseases, and other disorders. Uncoupling protein 2(UCP2) is the effector responsible for regulating cellular thermogenesis and ROS production via dissipating protons in an electrochemical gradient. A UCP2 inhibitor named genipin(GNP) is being researched for its effect on mitochondrial temperature, but little is known about its mechanisms. This study developed several molecular probes to explore the interactions between GNP and UCP2. The result indicated that the hemiacetal structure in GNP could selectively react with the ?-amine of lysine on the UCP2 proton leakage channel through ringopening condensation at the mitochondrial, cellular, and animal levels. A notable feature of the reaction is its temperature sensitivity and ability to conjugate with UCP2 at high fever as lysine-specific covalent inhibitors that prevent mitochondrial thermogenesis. The result not only clarifies the existence of an antipyretic properties of GNP via its irreversible coupling to UCP2, but also reveals a bioorthogonal reaction of hemiacetal iridoid aglycone for selectively binding with the ?-amine of lysine on proteins.展开更多
Chitosan (CS) microparticles with and without cross-linking were prepared by a water-in-oil emulsion solvent diffusion method without any surfactants. Aqueous CS solution and ethyl ace- tate were used as water and oil...Chitosan (CS) microparticles with and without cross-linking were prepared by a water-in-oil emulsion solvent diffusion method without any surfactants. Aqueous CS solution and ethyl ace- tate were used as water and oil phases, respectively. Genipin was used as a cross-linker. Influ- ences of genipin ratios and cross-linking times on CS microparticle characteristics were investigated. Non-cross-linked and cross-linked CS microparticles were spherical in shape and rough in surface. Microparticle matrices showed porous structures. Surface roughness, mean par- ticle sizes and bulk density of CS microparticles increased and their dissolutions in acetic acid solution decreased when genipin ratio and cross- linking time increased.展开更多
There is an increasing demand for crosslinking methods of silk fibroin (SF) scaffolds in biomedical applications that could maintain the biocompatibility, bioactivity as well as improve the water resistance and mechan...There is an increasing demand for crosslinking methods of silk fibroin (SF) scaffolds in biomedical applications that could maintain the biocompatibility, bioactivity as well as improve the water resistance and mechanical properties of SF materials. In this study, SF was crosslinked effectively with genipin which is a naturally occurring iridoid glucoside and the crosslinking mechanism was investigated through FTIR and amino acid analysis. The results showed that genipin could react with the -NH2 groups on the side chains of SF macromolecules and to form inter- and intra-molecular covalent bonds, and improved the stability of SF materials significantly. In vitro, the performances of genipin-crosslinked SF films were assessed by seeding L929 cells and compared with ethanol-processed SF films, glutaraldehyde and polyethylene glycol diglycidyl ether crosslinked ones. The genipin-crosslinked SF films showed a similar affinity to cells as ethanol-processed ones, and a higher bioactivity in promoting cell growth and proliferation, inhibition of cell apoptosis, and maintenance of normal cell cycle compared with glutaraldehyde and polyethylene glycol diglycidyl ether crosslinked SF films. These features, combined with the decrease of brittleness of SF films crosslinked with chemical methods, substantiated genipin as an effective and biocompatible agent for the manufacturing of bioactive SF materials which used as tissue engineering scaffolds and drug delivery carriers.展开更多
Polyvinyl alcohol gelatin hydrogels were fabricated using genipin as a crosslinking agent for bone regeneration application. Optimized formulation of PVA-GE hydrogel was fabricated using genipin as crosslinking agent....Polyvinyl alcohol gelatin hydrogels were fabricated using genipin as a crosslinking agent for bone regeneration application. Optimized formulation of PVA-GE hydrogel was fabricated using genipin as crosslinking agent. Characterizations such as FTIR, morphology, porosity, pore size, degradation and swelling rate were investigated. Bone regeneration potential of optimized genipin cross-linked polyvinyl alcohol-gelatin (PVA20) hydrogels was assessed by implanting in rabbit’s femur defect for 1, 5 and 15 weeks period. Results showed interconnected porosity as observed in scanning electron microscopy and successful crosslinking as confirmed by FTIR analysis. Increased porosity (92% ± 2.46%) and pore size distribution (100 - 200 μm) were also observed as well as decrease in swelling rate (426% ± 10.50%). Bone formation was evident in micro-CT after 5 and 15 days of in vivo implantation period. Micro-CT analysis showed 32.67% increased bone formation of PVA-GE hydrogel defect compared with negative control after 15 weeks of in-vivo implantation. Histological analyses showed no inflammatory reaction post implantation and increase in cell matrix formation after 5 and 15 weeks. The combined physical and chemical method of crosslinking promises improved mechanical properties of PVA-GE hydrogel making it a potential scaffold for bone tissue engineering applications.展开更多
The inflammation can be stimulated by the surgical implantation and biomaterial presence through the foreign body via bio-interface.Macrophages play a key role in the interaction of host tissue to implant surfaces.In ...The inflammation can be stimulated by the surgical implantation and biomaterial presence through the foreign body via bio-interface.Macrophages play a key role in the interaction of host tissue to implant surfaces.In present study,the immuno-inflammatory responses of genipin crosslinked gelatin matrices(GCGM)to macrophages in vitro and the host tissue surfaces in rats were investigated.The results showed that the mechanical properties,swelling and degradation of gelatin matrices were improved by the crosslinking of genipin at physiological conditions.The macrophage on the surface of GCGM could avoid to be activated.The interaction of macrophage and GCGM suggested that GCGM could reduce the inflammatory response with downregulating the production and m RNA expression of pro-inflammatory cytokines.The anti-inflammatory effect of GCGM was demonstrated to be related to inhibit nuclear factor kappa-B(NF-κB)signaling pathway.Furthermore,gelatin matrices crosslinked with genipin could decrease the acute and chronic inflammatory response interacting with host tissue surfaces to enhance the biocompatibility in rats.These results showed that GCGM could avoid to active macrophages and were endowed with anti-inflammatory properties,suggesting the significant potential for clinical success with the development of immunomodulatory biomaterials.展开更多
The study used a naturally occurring crosslinking reagent-genipin to chemically modify acellular bovine pericardium, prepare cardiac valve tissue engineering scaffold material,and evaluated genipin crosslinked acellul...The study used a naturally occurring crosslinking reagent-genipin to chemically modify acellular bovine pericardium, prepare cardiac valve tissue engineering scaffold material,and evaluated genipin crosslinked acellular matrix of bovine pericardium by investigating the physical and chemical properties of the tissues, such as the surface properties, crosslinking characteristics, mechanical properties, resistance to enzymatic capacity in vitro, and hemolysis tests. The results showed that acellular bovine pericardium matrix erosslinked with genipin was strong hydrophilicity, high crosslinking index, and stable structure, which can maintain good mechanical properties. As a kind of scaffold material for valve tissue engineering, it has wide application prospect.展开更多
Objective To investigate the change rules of endogenous metabolites in pancreas of the diabetic rats, and to explore the mechanism of genipin treatment for diabetes rats. Methods Metabolomic method based on ^1H-NMR wa...Objective To investigate the change rules of endogenous metabolites in pancreas of the diabetic rats, and to explore the mechanism of genipin treatment for diabetes rats. Methods Metabolomic method based on ^1H-NMR was applied, the diabetic rat model was prepared by ip injecting alloxan, and the high-, mid-, and low-dose genipin or metformin hydrochloride was ig injected as well as the rats in control and model groups were given the same volume of normal saline for 2 weeks. The pancreases of rats were collected and ^1H-NMR test was conducted, the metabolomic technology was adopted to analyze the endogenous metabolite changes in pancreas. Results The high-dose genipin possessed a better hypoglycemic effect, which could increase the contents of isoleucine, phosphatidylcholine, and phosphatidylethanolamine, and significantly reduce the contents of lactic acid, alanine, glutamine, aspartic acid, and creatine in pancreas of diabetic rats. Conclusion This study provided a theoretical basis for further exploration on the pathogenesis of diabetes and the mechanism of genipin for treatment of diabetes.展开更多
Crosslinked biopolymers were prepared for controlled release to investigate the diffusion of ascorbic acid from swelling high-solid networks in neutral and acidic pH conditions.Polymer swelling was measured at pH 2.8 ...Crosslinked biopolymers were prepared for controlled release to investigate the diffusion of ascorbic acid from swelling high-solid networks in neutral and acidic pH conditions.Polymer swelling was measured at pH 2.8 and pH 6.8 and change in microstructural properties,including network mesh size and average molecular weight between crosslinks of genipin-crosslinked gelatin and genipin-crosslinked chitosan systems,were predicted.Variable mathematical modelling was applied to the first 60%of ascorbic acid diffusion and statistical analysis was used to determine the model that best fits the experimental data.Molecular transport from the gelatin-based networks was best described by the Peppas-Sahlin model whereas the distinct effect of pH on chitosan structure formation brought into consideration the zero-order model.Mathematical modelling of bioactive compound release from swelling biopolymer matrices at different pH conditions is essential for the design of efficient targeted delivery matrices for functional food and nutraceutical applications.展开更多
Current treatment strategies for diabetic retinopathy(DR),an eye condition that can lead to blindness,have mainly focused on proliferative DR,including vitreous injection,retinal photocoagulation,and vitrectomy.Vitreo...Current treatment strategies for diabetic retinopathy(DR),an eye condition that can lead to blindness,have mainly focused on proliferative DR,including vitreous injection,retinal photocoagulation,and vitrectomy.Vitreous injections mainly depend on anti-vascular endothelial growth factor therapy.In this editorial,we comment on the article by Sun et al.We focus specifically on the mechanisms of the protective effect of genipin on the retina.Genipin is a gardenia extract used in traditional Chinese medicine(TCM).In their study,the authors suggest that controlling advanced glycosylation by the intraocular injection of genipin may be a strategy for preventing retinopathy.The innovative use of a Chinese medicine extract injected into the eye to achieve a curative effect has attracted our attention.Although TCM is effective in treating DR,the topical application of DR,especially intraocular injections,is not yet feasible.Herein,we present a brief analysis of effective Chinese medicines for the treatment of DR.The effectiveness of local injections of TCM applied directly into the eyes holds promise as an effective treatment approach for DR.展开更多
基金supported by the National Natural Science Foundation of China(No.81441096)the Project of Science and Technology of Shanxi Province(201603D321077)+1 种基金the Key Laboratory of Shanxi Province(201605D111004)the Science and Technology Innovation Team of Shanxi Province(201605D131045-18)
文摘Diabetic nephropathy is one of the various complications of diabetes mellitus, affecting patients for lifetime. Earlier studies have revealed that genipin can not only improve diabetes, but also induce cytotoxicity. Therefore, it is not clear which effect of genipin on kidneys occurs, when it is used in the treatment of diabetes. In the present study, we performed nuclear magnetic resonance(NMR)-based metabolomics analysis of urine and kidney tissue samples obtained from diabetic rats to explore the change of endogenous metabolites associated with diabetes and concomitant kidney disease. Nine significant differential metabolites that were closely related to renal function were screened. They were mainly related to three metabolic pathways: synthesis and degradation of ketone bodies, glycine, serine and threonine metabolism, and butanoate metabolism, which are involved in methylamine metabolism, energy metabolism and amino acid metabolism. In addition, after the intervention of genipin, the metabolic levels of all the metabolites tended to be normal, indicating a protective effect of genipin on kidneys. Our results may be helpful for understanding the antidiabetic effect of genipin.
基金the National Natural Science Foundation of China,No.81870650,No.81570832,and No.81900885Science and Technology Program Chongqing,No.2018GDRC008 and No.XKTS049。
文摘BACKGROUND Glycation is an important step in aging and oxidative stress,which can lead to endothelial dysfunction and cause severe damage to the eyes or kidneys of diabetics.Inhibition of the formation of advanced glycation end products(AGEs)and their cell toxicity can be a useful therapeutic strategy in the prevention of diabetic retinopathy(DR).Gardenia jasminoides Ellis(GJE)fruit is a selective inhibitor of AGEs.Genipin is an active compound of GJE fruit,which can be employed to treat diabetes.AIM To confirm the effect of genipin,a vital component of GJE fruit,in preventing human retinal microvascular endothelial cells(hRMECs)from AGEs damage in DR,to investigate the effect of genipin in the down-regulation of AGEs expression,and to explore the role of the CHGA/UCP2/glucose transporter 1(GLUT1)signal pathway in this process.METHODS In vitro,cell viability was tested to determine the effects of different doses of glucose and genipin in hRMECs.Cell Counting Kit-8(CCK-8),colony formation assay,flow cytometry,immunofluorescence,wound healing assay,transwell assay,and tube-forming assay were used to detect the effect of genipin on hRMECs cultured in high glucose conditions.In vivo,streptozotocin(STZ)induced mice were used,and genipin was administered by intraocular injection(IOI).To explore the effect and mechanism of genipin in diabetic-induced retinal dysfunction,reactive oxygen species(ROS),mitochondrial membrane potential(MMP),and 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose(2-NBDG)assays were performed to explore energy metabolism and oxidative stress damage in high glucose-induced hRMECs and STZ mouse retinas.Immunofluorescence and Western blot were used to investigate the expression of inflammatory cytokines[vascular endothelial growth factor(VEGF),SCG3,tumor necrosis factor-alpha(TNF-α),interleukin(IL)-1β,IL-18,and nucleotide-binding domain,leucine-rich-containing family,pyrin domain-containing 3(NLRP3)].The protein expression of the receptor of AGEs(RAGE)and the mitochondria-related signal molecules CHGA,GLUT1,and UCP2 in high glucose-induced hRMECs and STZ mouse retinas were measured and compared with the genipin-treated group.RESULTS The results of CCK-8 and colony formation assay showed that genipin promoted cell viability in high glucose(30 mmol/L D-Glucose)-induced hRMECs,especially at a 0.4μmol/L dose for 7 d.Flow cytometry results showed that high glucose can increase apoptosis rate by 30%,and genipin alleviated cell apoptosis in AGEs-induced hRMECs.A high glucose environment promoted ATP,ROS,MMP,and 2-NBDG levels,while genipin inhibited these phenotypic abnormalities in AGEs-induced hRMECs.Furthermore,genipin remarkably reduced the levels of the pro-inflammatory cytokines TNF-α,IL-1β,IL-18,and NLRP3 and impeded the expression of VEGF and SCG3 in AGEs-damaged hRMECs.These results showed that genipin can reverse high glucose induced damage with regard to cell proliferation and apoptosis in vitro,while reducing energy metabolism,oxidative stress,and inflammatory injury caused by high glucose.In addition,ROS levels and glucose uptake levels were higher in the retina from the untreated eye than in the genipin-treated eye of STZ mice.The expression of inflammatory cytokines and pathway protein in the untreated eye compared with the genipin-treated eye was significantly increased,as measured by Western blot.These results showed that IOI of genipin reduced the expression of CHGA,UCP2,and GLUT1,maintained the retinal structure,and decreased ROS,glucose uptake,and inflammation levels in vivo.In addition,we found that SCG3 expression might have a higher sensitivity in DR than VEGF as a diagnostic marker at the protein level.CONCLUSION Our study suggested that genipin ameliorates AGEs-induced hRMECs proliferation,apoptosis,energy metabolism,oxidative stress,and inflammatory injury,partially via the CHGA/UCP2/GLUT1 pathway.Control of advanced glycation by IOI of genipin may represent a strategy to prevent severe retinopathy and vision loss.
基金Supported by Science and Technology Projects of Guangdong Province,China(No.2007B031002001,No.2008B030301086)
文摘·AIM: To study the feasibility of using the discoloration to evaluate the biomechanical properties after treating with genipin.·METHODS: Porcine cadaver eyes were treated for30 min with 1.0%(by w/v) genipin. Untreated samples were used as controls. After treatment, scleral strips of4.0 ×10.0-mm2 were cut. The denaturation temperature(Td) measurement and stress-strain test were performed after taking photograph to analyze the color.·RESULTS: Within 24 h after treating with genipin, the sclera exhibited a bluish color which became deeper with time. And the denaturation temperature also was increased gradually. Compared with untreated groups, at1, 6, 12, 24 and 36 h after treatment, the ultimate stress were increased by 56%, 153%, 173%, 225% and 211%respectively. The Young’s modulus at 10% strain also increased by 170%, 246%, 264%, 389% and 288%respectively. There were strong correlation between the discoloration and the biomechanical properties(ΔE-Ultimate stress:R2=0.892, P =0.00; ΔE-Young’s modulus:R2=0.602, P =0.00).·CONCLUSION: Genipin could be used to strengthen collagen gradually in a relatively short time span. And the biomechanical properties could be reliably evaluated via simple visible discoloration.
基金National Natural Science Foundation of China(Grant No.82174067,81960646 and 82004080)。
文摘In the present study,we aimed to ascertain the metabolic detoxification routes of genipin via CYP3A4,SULT2 A1,and UGT1 A1 in Hepa RG cells.It was found that the hepatic CYP3A4,SULT2 A1,and UGT1 A1 synergistically mediated the metabolic detoxification of genipin,and the CYP3A4 was the limited enzyme.In detail,the pivotal detoxification pathway was CYP3A4-SULT2 A1/UGT1 A1,indicating that SULT2 A1 and UGT1 A1 further catalyzed the phase II detoxification metabolism followed by the genipin metabolization by CYP3A4 to the phase I metabolites with alleviated toxicity.Our findings provided valuable cues for future studies on the detoxification of genipin,even the compatibility detoxification of Zhi-zi.Moreover,these data facilitated the development and rational administration of genipin and Zhi-zi.
基金Supported by Guangdong Province Science and Technology Projects(No.2007B031002001)
文摘AIM：To strengthen the biomechanics of collagen by crosslinking rabbit scleral collagen with genipin to develop a new therapy for preventing myopic progression. METHODS：Ten New Zealand rabbits were treated with 0.5 mmol/L genipin injected into the sub-Tenon＇s capsule in the right eyes. Untreated contralateral eyes served as the control. The treated area was cut into scleral strips measuring 4.0 mm×10.0 mm for stress-strain measurements（n=5）. The remaining five treated eyes were prepared for histological examination.RESULTS：Compared to the untreated scleral strips,the genipin-crosslinked scleral strips showed that the ultimate stress and Young＇s modulus at 10% strain were increased by the amplitude of 130% and 303% respectively,ultimate strain was decreased by 24%. There had no α-smooth muscle actin（α-SMA）positive cells in control and treated sclera. Histologically,there was no sign of apoptosis in the sclera,choroid,and retina; and no side effects were found in the peripheral cornea and optic nerve adjacent to the treatment area.CONCLUSION：Genipin induced crosslinking of collagen can increase its biomechanical behavior by direct strengthening of the extracellular matrix in rabbit sclera,with no α-SMA expression seen in the myofibroblasts. As there is no evidence of cytotoxicity in the scleral,choroidal,and retinal cells,genipin is likely a promising agent to strengthen the weakened sclera to prevent myopic progression.
文摘Antimicrobial delivery has been advocated for guided tissue regeneration (GTR) or guided bone regeneration (GBR) therapies involving patients with aggressive or unresolved periodontitis/peri-implantitis. Electrospun chitosan membranes demonstrate several advantages over traditional GTR barrier membranes because they stimulate healing, mimic the topology of the extracellular matrix, and allow for diffusion of nutrients and wastes into/out of the graft site, and were shown to stimulate bone formation in a rabbit calvarial criticalsize defect model. Previously, we have shown improvements in mechanical properties and degradation kinetics by crosslinking electrospun membranes with 5 mM or 10 mM genipin. We have also demonstrated the ability of elecrospun chitosan membranes to inhibit lippopolysaccharide (LPS)-induced monocyte activation. In this study, minocycline was incorporated into the chitosan membrane by passive absorption at 5 or 10 mg/mL. The minocycline-loaded membranes and control membranes (carrier only) were tested against Porphyromonas gingivalis (P. gingivalis) by repeated zone of inhibition (ZOI) measurements. Testing showed that uncrosslinked and genipin-crosslinked membranes have similar capacity to absorb aqueous solutions (swelling ratio 1.7 - 2.2). Minocycline loading resulted in bacterial inhibition for up to 8 days from crosslinked membranes (with 11 mm initial ZOI) whereas uncrosslinked membranes loaded with minocycline only inhibited bacteria for 4 days (with 8 mm initial ZOI). These in vitro results suggest that genipin-crosslinked electrospun chitosan membranes loaded with minocycline may be able to reduce early bacterial contamination of GTR graft sites.
基金supported by the National Natural Science Foundation of China(No.81973449)the National Key Research and Development Program of China(Nos.2018YFC1704800 and 2018YFC1704805)。
文摘Mitochondria are essential for eukaryotic life as powerhouses for energy metabolism. Excessive mitochondrial hyperthermia and reactive oxygen species(ROS) production have been associated with aging, cancer,neurodegenerative diseases, and other disorders. Uncoupling protein 2(UCP2) is the effector responsible for regulating cellular thermogenesis and ROS production via dissipating protons in an electrochemical gradient. A UCP2 inhibitor named genipin(GNP) is being researched for its effect on mitochondrial temperature, but little is known about its mechanisms. This study developed several molecular probes to explore the interactions between GNP and UCP2. The result indicated that the hemiacetal structure in GNP could selectively react with the ?-amine of lysine on the UCP2 proton leakage channel through ringopening condensation at the mitochondrial, cellular, and animal levels. A notable feature of the reaction is its temperature sensitivity and ability to conjugate with UCP2 at high fever as lysine-specific covalent inhibitors that prevent mitochondrial thermogenesis. The result not only clarifies the existence of an antipyretic properties of GNP via its irreversible coupling to UCP2, but also reveals a bioorthogonal reaction of hemiacetal iridoid aglycone for selectively binding with the ?-amine of lysine on proteins.
文摘Chitosan (CS) microparticles with and without cross-linking were prepared by a water-in-oil emulsion solvent diffusion method without any surfactants. Aqueous CS solution and ethyl ace- tate were used as water and oil phases, respectively. Genipin was used as a cross-linker. Influ- ences of genipin ratios and cross-linking times on CS microparticle characteristics were investigated. Non-cross-linked and cross-linked CS microparticles were spherical in shape and rough in surface. Microparticle matrices showed porous structures. Surface roughness, mean par- ticle sizes and bulk density of CS microparticles increased and their dissolutions in acetic acid solution decreased when genipin ratio and cross- linking time increased.
文摘There is an increasing demand for crosslinking methods of silk fibroin (SF) scaffolds in biomedical applications that could maintain the biocompatibility, bioactivity as well as improve the water resistance and mechanical properties of SF materials. In this study, SF was crosslinked effectively with genipin which is a naturally occurring iridoid glucoside and the crosslinking mechanism was investigated through FTIR and amino acid analysis. The results showed that genipin could react with the -NH2 groups on the side chains of SF macromolecules and to form inter- and intra-molecular covalent bonds, and improved the stability of SF materials significantly. In vitro, the performances of genipin-crosslinked SF films were assessed by seeding L929 cells and compared with ethanol-processed SF films, glutaraldehyde and polyethylene glycol diglycidyl ether crosslinked ones. The genipin-crosslinked SF films showed a similar affinity to cells as ethanol-processed ones, and a higher bioactivity in promoting cell growth and proliferation, inhibition of cell apoptosis, and maintenance of normal cell cycle compared with glutaraldehyde and polyethylene glycol diglycidyl ether crosslinked SF films. These features, combined with the decrease of brittleness of SF films crosslinked with chemical methods, substantiated genipin as an effective and biocompatible agent for the manufacturing of bioactive SF materials which used as tissue engineering scaffolds and drug delivery carriers.
文摘Polyvinyl alcohol gelatin hydrogels were fabricated using genipin as a crosslinking agent for bone regeneration application. Optimized formulation of PVA-GE hydrogel was fabricated using genipin as crosslinking agent. Characterizations such as FTIR, morphology, porosity, pore size, degradation and swelling rate were investigated. Bone regeneration potential of optimized genipin cross-linked polyvinyl alcohol-gelatin (PVA20) hydrogels was assessed by implanting in rabbit’s femur defect for 1, 5 and 15 weeks period. Results showed interconnected porosity as observed in scanning electron microscopy and successful crosslinking as confirmed by FTIR analysis. Increased porosity (92% ± 2.46%) and pore size distribution (100 - 200 μm) were also observed as well as decrease in swelling rate (426% ± 10.50%). Bone formation was evident in micro-CT after 5 and 15 days of in vivo implantation period. Micro-CT analysis showed 32.67% increased bone formation of PVA-GE hydrogel defect compared with negative control after 15 weeks of in-vivo implantation. Histological analyses showed no inflammatory reaction post implantation and increase in cell matrix formation after 5 and 15 weeks. The combined physical and chemical method of crosslinking promises improved mechanical properties of PVA-GE hydrogel making it a potential scaffold for bone tissue engineering applications.
基金supported by the National Natural Science Foundation of China(81741119)Fundamental Research Funds for the Central Universities,China(3332020059)。
文摘The inflammation can be stimulated by the surgical implantation and biomaterial presence through the foreign body via bio-interface.Macrophages play a key role in the interaction of host tissue to implant surfaces.In present study,the immuno-inflammatory responses of genipin crosslinked gelatin matrices(GCGM)to macrophages in vitro and the host tissue surfaces in rats were investigated.The results showed that the mechanical properties,swelling and degradation of gelatin matrices were improved by the crosslinking of genipin at physiological conditions.The macrophage on the surface of GCGM could avoid to be activated.The interaction of macrophage and GCGM suggested that GCGM could reduce the inflammatory response with downregulating the production and m RNA expression of pro-inflammatory cytokines.The anti-inflammatory effect of GCGM was demonstrated to be related to inhibit nuclear factor kappa-B(NF-κB)signaling pathway.Furthermore,gelatin matrices crosslinked with genipin could decrease the acute and chronic inflammatory response interacting with host tissue surfaces to enhance the biocompatibility in rats.These results showed that GCGM could avoid to active macrophages and were endowed with anti-inflammatory properties,suggesting the significant potential for clinical success with the development of immunomodulatory biomaterials.
基金National High-tech Research and Development Program(836 Prgoram) of Chinagrant number:2007AA02Z444+1 种基金National BasicProgram of China (973 Program)grant number:2009CB930000
文摘The study used a naturally occurring crosslinking reagent-genipin to chemically modify acellular bovine pericardium, prepare cardiac valve tissue engineering scaffold material,and evaluated genipin crosslinked acellular matrix of bovine pericardium by investigating the physical and chemical properties of the tissues, such as the surface properties, crosslinking characteristics, mechanical properties, resistance to enzymatic capacity in vitro, and hemolysis tests. The results showed that acellular bovine pericardium matrix erosslinked with genipin was strong hydrophilicity, high crosslinking index, and stable structure, which can maintain good mechanical properties. As a kind of scaffold material for valve tissue engineering, it has wide application prospect.
文摘Objective To investigate the change rules of endogenous metabolites in pancreas of the diabetic rats, and to explore the mechanism of genipin treatment for diabetes rats. Methods Metabolomic method based on ^1H-NMR was applied, the diabetic rat model was prepared by ip injecting alloxan, and the high-, mid-, and low-dose genipin or metformin hydrochloride was ig injected as well as the rats in control and model groups were given the same volume of normal saline for 2 weeks. The pancreases of rats were collected and ^1H-NMR test was conducted, the metabolomic technology was adopted to analyze the endogenous metabolite changes in pancreas. Results The high-dose genipin possessed a better hypoglycemic effect, which could increase the contents of isoleucine, phosphatidylcholine, and phosphatidylethanolamine, and significantly reduce the contents of lactic acid, alanine, glutamine, aspartic acid, and creatine in pancreas of diabetic rats. Conclusion This study provided a theoretical basis for further exploration on the pathogenesis of diabetes and the mechanism of genipin for treatment of diabetes.
基金This research is supported by an Australian Government Research Training Program(RTP)Scholarship to Felicity Whitehead.
文摘Crosslinked biopolymers were prepared for controlled release to investigate the diffusion of ascorbic acid from swelling high-solid networks in neutral and acidic pH conditions.Polymer swelling was measured at pH 2.8 and pH 6.8 and change in microstructural properties,including network mesh size and average molecular weight between crosslinks of genipin-crosslinked gelatin and genipin-crosslinked chitosan systems,were predicted.Variable mathematical modelling was applied to the first 60%of ascorbic acid diffusion and statistical analysis was used to determine the model that best fits the experimental data.Molecular transport from the gelatin-based networks was best described by the Peppas-Sahlin model whereas the distinct effect of pH on chitosan structure formation brought into consideration the zero-order model.Mathematical modelling of bioactive compound release from swelling biopolymer matrices at different pH conditions is essential for the design of efficient targeted delivery matrices for functional food and nutraceutical applications.
基金Supported by National Natural Science Foundation of China,No.82104862Scientific Research Project Foundation of Zhejiang Chinese Medical University,No.2023FSYYZZ01 and No.2023RCZXZK49.
文摘Current treatment strategies for diabetic retinopathy(DR),an eye condition that can lead to blindness,have mainly focused on proliferative DR,including vitreous injection,retinal photocoagulation,and vitrectomy.Vitreous injections mainly depend on anti-vascular endothelial growth factor therapy.In this editorial,we comment on the article by Sun et al.We focus specifically on the mechanisms of the protective effect of genipin on the retina.Genipin is a gardenia extract used in traditional Chinese medicine(TCM).In their study,the authors suggest that controlling advanced glycosylation by the intraocular injection of genipin may be a strategy for preventing retinopathy.The innovative use of a Chinese medicine extract injected into the eye to achieve a curative effect has attracted our attention.Although TCM is effective in treating DR,the topical application of DR,especially intraocular injections,is not yet feasible.Herein,we present a brief analysis of effective Chinese medicines for the treatment of DR.The effectiveness of local injections of TCM applied directly into the eyes holds promise as an effective treatment approach for DR.
