Exploring the material basis of genipin-induced hepatotoxicity in vitro / 药学学报

The current study explored the hepatotoxicity among closed-ring genipin, open-ring tautomer of genipin and gardenia blue that generated from genipin and amino acid reaction using HepaRG cells to identify the material basis of genipin-induced hepatotoxicity in vitro. The effects of temperature, pH value and different kinds of amino acids on the chemical structure tautomerism between closed-ring and open-ring tautomer of genipin and the production of gardenia blue were investigated firstly, which aimed to explicit the conditions that could distinguish the closed-ring genipin and its open-ring tautomer, and the conditions generating gardenia blue, which were applied to prepare different kinds of gardenia blue; the CCK-8 kit was employed to analyze the hepatotoxicity of closed-ring genipin, open-ring tautomer of genipin and gardenia blue. From the results, it was found that, the structure transformation from close-ring to open-ring of genipin could be inhibited under the condition with acid environment; being essential groups to generate gardenia blue, the primary amino group and the open-ring tautomer of genipin reacting to generate the dihydropyridine ring was probably the key structure of gardenia blue; the structure characteristics existed apparent distinction at the reactive temperature of 37 ℃ and 80 ℃; compared to the culture condition with pH = 7.4, the concentration of genipin with close-ring in culture medium was significantly increased at pH = 5, but the cell viability did not decreased; the cell toxicity of gardenia blue was apparently lower than open-ring tautomer of genipin, and even some kinds of gardenia blue showed growth promoting effect on HepaRG cells. Here, it was suggested potentially that open-ring tautomer of genipin be the important material basis to induce hepatotoxicity, which could provide a cue and lay a foundation for the elucidation of the underlying mechanism of genipin-induced hepatotoxicity.

Advances in the efficacy and safety of scleral collagen cross-linking in the prevention and treatment of pathologic myopia / 国际眼科杂志(Guoji Yanke Zazhi)

In 2004, it was the first time that Wollensak and Spoerl had applied physical and chemical cross-linking methods to scleral tissue. They found that the biomechanical strength of cross-linked sclera, induced by riboflavin/ultraviolet A, glyceraldehyde and glutaraldehyde, could be improved and proposed that scleral collagen cross-linking is expected to be a new method for the treatment of pathologic myopia. In recent years, a series of explorations have been made on the effectiveness and adverse reactions of physical and chemical cross-linking in the prevention and treatment of pathologic myopia, including the establishment of various animal models and different myopia modeling methods, the improvement of cross-linking methods, the amelioration of the measurement of biomechanical strength of scleral tissue and the attention of biological parameters such as the thickness of retinal nerve fiber layer and the amplitude of electroretinogram in vivo. Genipin-crosslinking of the scleral collagen combined with posterior scleral contraction/reinforcement has been applied to clinical research. This review summarizes physical cross-linking and the genipin-crosslinking of scleral collagen to explore the effectiveness and safety of the methods in the prevention and treatment of the pathologic myopia.

Preparation and detection of VEGF chitosan microspheres with different concentrations / 中华医学美学美容杂志

Objective To evaluate the effects of chitosan microspheres as the ball wall,pre pared by emulsion cross-linked VEGF chitosan microspheres.Methods 1 ％ chitosan aqueous solution as the aqueous solution and genipin as a cross-linking agent were used to detect the drug loading rate and cumulative release rate of drug-loaded microspheres by ELISA.Results VEGF chitosan microsphere size was about 5.25-20.34 μm;electron microscopy showed that microspheres surface was smooth,decentralized,and had small adhesions each other.The highest encapsulation efficiency was (56.33±3.42) ％ and the highest drug loading rate was (18.75±0.32) ng/mg.In vitro release experiments showed that the concentration of VEGF increased gradually with the passage of time.The release rate of microspheres was (32.32±2.31)％ on the first day and (86.42±1.68)％ on the 21st day.The cumulative release rate was 88.7％.Conclusions Genipin can be used to prepare chitosan microspheres loaded with different concentrations of VEGF with good performance and simple preparation process.The drug-loading rate and entrapment efficiency of the obtained microspheres are both high.

A Comparative Study of the Effects of Different Decellularization Methods and Genipin-Cross-Linking on the Properties of Tracheal Matrices

BACKGROUND: Different decellularization methods can affect the integrity and the biomechanical and biocompatible properties of the tracheal matrix. Natural cross-linking with genipin can be applied to improve those properties. The goals of this study were to evaluate the effects of different decellularization methods on the properties of genipin-cross-linked decellularized tracheal matrices in rabbits. METHODS: The tracheas of New Zealand rabbits were decellularized by the Triton-X 100-processed method (TPM) and the detergent-enzymatic method (DEM) and were then cross-linked with genipin. Mechanical tests, haematoxylin-eosin staining, Masson trichrome staining, Safranin O staining, DAPI staining, scanning electronic microscopy (SEM), and biocompatibility tests were used to evaluate the treatment. The bioengineered trachea and control trachea were then implanted into allogeneic rabbits for 30 days. The structural and functional analyses were performed after transplantation. RESULTS: The biomechanical tests demonstrated that the biomechanical properties of the decellularized tracheas decreased and that genipin improved them (p < 0.05). The histological staining results revealed that most of the mucosal epithelial cells were removed and that the decellularized trachea had lower immunogenicity than the control group. The analysis of SEM revealed that the decellularized trachea retained the micro- and ultra-structural architectures of the trachea and that the matrices cross-linked with genipin were denser. The biocompatibility evaluation and in vivo implantation experiments showed that the decellularized trachea treated with the DEM had better biocompatibility than that treated with the TPM and that immunogenicity in the cross-linked tissues was lower than that in the uncross-linked tissues (p < 0.05). CONCLUSION: Compared with the trachea treated with the TPM, the rabbit trachea processed by the DEM had better biocompatibility and lower immunogenicity, and its structural and mechanical characteristics were effectively improved after the genipin treatment, which is suitable for engineering replacement tracheal tissue.

Genipin Inhibits Hypoxia-Induced Accumulation of HIF-1α and VEGF Expressions in Human Cervical Carcinoma Cells / 고신대학교의과대학학술지

OBJECTIVES: Hypoxia—a characteristic of almost all types of solid tumors—has been associated with poor outcomes in several human malignancies. Genipin—an active constituent of Gardenia fruit— has been reported to exert an anti-tumor effect in several cancers. In this study, we investigated inhibition of angiogenesis using Genipin-mediated hypoxia-induced hypoxia inducible factor (HIF-1) and VEGF expression in human cervical cancer cells.METHODS: Under normoxic and hypoxic conditions, the expression of HIF-1α and VEGF in cervical cancer HeLa cells was detected by quantitative reverse transcription polymerase chain reaction and western blotting. Luciferase reporter assays were used to investigate the molecular mechanisms underlying the hypoxia-induced survivin activation.RESULTS: Surprisingly, we found that Genipin suppressed the HIF-1α accumulation during hypoxia in human liver cancer cell line (HepG2), human prostate cancer cell line (LNCaP), colon cancer cell line (HCT116), and breast cancer cell line (MDA231). Genipin treatment also significantly reduced hypoxia-induced secretion of VEGF.CONCLUSIONS: Suppression of HIF-1α accumulation following treatment with Genipin under hypoxia was associated with PI3K and MAPK pathways. Taken together, these results suggested that Genipin inhibits HIF-1α expression through inhibition of PI3K and MAPK signaling pathways. These results provide new insights into a potential mechanism of the anticancer properties of Genipin.

Genipin cross-linked decellularized scaffold for allogenic transplantation in situ / 国际生物医学工程杂志

Objective@#To explored the bio-compatibility and cartilage regeneration of the rabbits genipin cross-linked decellularized scaffold, to provide experimental and theoretical support for the clinical application of genipin cross-linked decellularized scaffold.@*Methods@#Detergent-enzyme method was used to prepare decellularized tracheal scaffolds. Cellular content of native trachea and decellularized trachea were compared by 4′, 6-diamidino-2-phenylindole(DAPI) staining. Masson trichrome staining was used to compare the histological structure of the progenitor tube, decellularized trachea, and genipin cross-linked decellularized trachea. Nine adult New Zealand white rabbits were randomly divided into autologous tracheal transplantation group (negative control group), allogeneic tracheal transplantation group (positive control group), and genipin cross-linked decellularized tracheal transplantation group (experimental group). Autologous bone marrow mesenchymal stem cells were implanted on the surface of trachea in each group. The blood cells and type II collagen were detected to compare the inflammatory response and chondrocyte regeneration after tracheal orthotopic transplantation in the three groups.@*Results@#After DAPI staining and light microscope observation (×200), the cell content of the acellular 7-cycle trachea [(143.0 ± 71.1) cells/field] was significantly lower than that of the native trachea [(853.5 ± 149.6) cells/field], and the difference was statistically significant (P<0.001). Masson′s trichrome staining showed that the tissue structure of genipin cross-linked decellularized trachea was more complete. Blood cell analysis and type II collagen test results showed that genipin cross-linked decellularized trachea transplanted with bone marrow mesenchymal stem cells after transplantation in situ has little rejection and can be converted into chondrocytes by the action of related growth factors in vivo.@*Conclusions@#Genipin cross-linked decellularized tracheal scaffold combined with stem cell transplantation can successfully construct a tracheal in situ replacement model. This study provides a strong support for the research of tissue engineering trachea.

Quality evaluation of Gardeniae Fructus from different areas by using chemical pattern recognition based on combination of chromatographic fingerprints and multi-component content / 中草药

Objective: To evaluate the quality of Gardeniae Fructus from different producing areas and optimize the best place of production by using chemical pattern recognition method based on HPLC fingerprint and multi-component content determination. Methods: The RP-HPLC using acetonitrile-0.1% phosphoric acid aqueous solution as mobile phase based on the wavelength switching (234 nm and 440 nm) and gradient elution method was developed to evaluate the quality of 30 batches of Gardeniae Fructus from 10 different producing areas. The combination of chromatographic fingerprints and quality determination of five active ingredients, as well as chemometrics including hierarchical cluster analysis (HCA) and principal component analysis (PCA), were further employed for the quality assessment. Results: The fingerprint similarity of 30 batches of Gardeniae Fructus from 10 different producing areas were all above 0.95. The content ranges of deacetylated methyl oxalate, genipin gentiobioside, geniposide, crocin I, and crocin II were 0.61-4.26 mg/g, 1.73-12.92 mg/g, 51.79-82.76 mg/g, 5.03-12.80 mg/g, and 0.71-2.28 mg/g. Samples can be divided into three categories by HCA: The first category is Yongzhou in Hunan, Yong’ an in Hunan, Fengcheng in Jiangxi, Hukou in Jiangxi, Guangde in Anhui, and Fuding in Fujian; The second category is Lichuan in Jiangxi, Jurong in Jiangsu and Tanghe in Henan; And the third category is Lu’ an in Anhui. PCA was used to further evaluate the quality differences of the samples from different producing areas, and it was found that the medicinal materials from Lu’ an in Anhui, Hukou in Jiangxi, and Yong’ an in Hunan were of superior quality. Conclusion: The quality of Gardeniae Fructus from different producing areas was different to a certain extent, while the quality of same batches was stable. By combination of fingerprint and content determination, the chemical pattern recognition method can be used to evaluate the quality of the Gardeniae Fructus comprehensively. The establishment of this method provided an effective reference for the quality control and evaluation of Gardeniae Fructus.

Anti-cerebral ischemia mechanisms of baicalein and genipin based on network pharmacology / 中草药

Objective: Using network pharmacology and molecular docking technology along with scutellarein (SE) as a reference, this study predicted the anti-cerebral ischemia targets of both baicalein (BE) and genipin (GE). It is hoped that these will provide a reference for clinical prevention and development of ischemic diseases. Methods: SE, BE and GE targets were predicted using TCMSP, Swiss Target Prediction, Stitch database search and literature mining methods. Targets related to cerebral ischemia diseases could be predicted by DisGeNET, CTD, NCBI Gene, OMIM, DrugBank and PharmGkb databases. Cytoscape 3.3.0 was used to construct the small molecule-target network. GO function enrichment and KEGG pathway analysis of SE, BE and GE specific anti-cerebral ischemia targets were analyzed with the DAVID database. Autodock Vina software was used for molecular docking, testing the binding energy of BE, GE and SE to targets of cerebral ischemia. The optimal target protein was selected according to the binding energy and inhibition concentration of receptor and ligand. Results: A total of 30 potential targets of SE, 59 potential targets of BE and 35 potential targets of GE were found. Common anti-cerebral ischemia targets of SE and BE were PIK3CG, CYP1A2, VEGFA, ALOX5 and PTGS2, while common anti-cerebral ischemia targets of SE and GE were PTGS2. Molecular docking results demonstrated that the binding energy and inhibitory concentration of receptor PTGS2 to the three drugs were relatively low. Enrichment of GO function showed that common targets of BE-SE were mainly distributed in cytoplasm, organelle membrane, endoplasmic reticulum and other elements. These elements had binding functions with metal ions and cations, catalytic and oxidoreductase activities, and they participated in cell lipid, carboxylic acid, oxygenic acid, organic acid metabolism and fatty acid synthesis. Results: of the KEGG analysis demonstrated that receptor PTGS2 mainly acted on the arachidonic acid metabolism pathway and the vascular endothelial growth factor signaling pathway. A combination of BE and GE functioned in the treatment of cerebral ischemia disease by inhibiting expression of PTGS2 (COX-2) and vascular endothelial growth factor protein, thus reducing brain injury caused by inflammatory factors and improving the permeability of the blood brain barrier (BBB). Conclusion: This study predicted potential targets of BE and GE compatibility in the treatment of cerebral ischemia diseases and preliminarily verified mechanisms of action in the treatment of cerebral ischemia diseases. It provides valuable data for further study into the mechanisms of BE and GE compatibility for the treatment of cerebral ischemia as well as developing a basis for the next synthesis of new derivatives.

Inhibitory effect of scleral crosslinking using genipin on form-deprivation myopia in rabbits / 中华实验眼科杂志

Objective@#To investigate the effects of scleral crosslinking using genipin on ocular biological parameters and scleral biomechanics of form-deprivation myopia rabbits.@*Methods@#Sixty healthy New Zealand rabbits of 14 days old were collected.The right eyes were selected as experimental eye.The rabbits were randomly divided into three groups: control group with no treatment; myopia model group with eyelid suture procedure performed on the right eye; genipin injection group with eyelid suture procedure performed on the right eye combined with subconjunctival injection of genipin.The suture was removed 60 days after the eyelid suture procedure.The diopter, length of vitreous cavity, and axial length were measured.The sclera at 1: 00 and 7: 00 position of the experimental eye was used to make a scleral strip.The thickness, elastic modulus, creep rate, ultimate stress and ultimate strain of the sclera were measured.This study was approved by the animal experimental Ethics Committee of Bethune International Peace Hospital (2018-ky-09).@*Results@#The diopters of genipin injection group, myopia model group and control group were (2.50±1.38), (0.33±0.52) and (2.08±0.52)D, respectively, the axial lengths of the three groups were (15.33±0.82), (15.83±0.41) and (15.00±0.43)mm, respectively; the changes in vitreous cavity lengths were (1.50±0.79), (2.59±0.83) and (1.48±0.66)mm, respectively; and the ratios of vitreous cavity length to axial length were 0.46±0.02, 0.51±0.02 and 0.47±0.02, respectively.The diopter in myopia model group was significantly lower than those in control group and genipin injection group, the axial length in myopia model group was significantly longer than that in control group, the change in vitreous cavity lengths and ratio of vitreous cavity length to axial length in myopia model group were significantly higher than those in control group and genipin injection group, the axial length in myopia model group was significantly longer than that in control group, the differences were statistically significant (all at P<0.05). The elasticity modulus was (9.10±3.12)MPa and ultimate stress was (1.42±0.57)MPa in genipin injection group, which were significantly higher than (6.98±3.30)MPa and (1.15±0.57)MPa in control group, and (6.25±3.17)MPa and (0.82±0.38)MPa in myopia model group (all at P<0.05). The ultimate strain was (20.99±5.60)% in genipin injection group, which was significantly lower than (25.08±6.35)% in control group and (27.78±8.20)% in myopia model group, with significant differences between them (both at P<0.05).@*Conclusions@#Posterior sub-Tenon capsule injection of genipin for collagen crosslinking in the sclera increases the biomechanical strength of sclera and effectively prevents the development of form-deprivation myopia in rabbits, which provides a new idea for the prevention and treatment of myopia in the future.

A sensitive LC-MS/MS method for simultaneous assay of three iridiods in Gardenia jasminoides Ellis / 药学实践杂志

Objective To develop a simple, rapid and sensitive method for the determination of major compounds from Gardenia jasminoides Ellis using liquid chromatography tandem mass spectrometry (LC-MS/MS).Methods The analysis was performed on Dikma Diamonsil?C18 (100mm×4.6mm, 5μm) column with acetonitrile-0.1％acetic acid and 0.1％acetic acidwater as mobile phase at a rate of 0.4ml/min.The column temperature was set at 40℃and the injection volume was 2μl.Quantification of these compounds was performed by LC-MS/MS with positive or negative ion mode electrospray ionization (ESI) in the multiple reaction monitoring (MRM) mode.Nebulizer gas, 3L/h;drying gas, 15L/h;desolvation line (DL) temperature, 240℃;heat block temperature, 300℃;CID, 230kPa.The mass transition of the precursor/product ions was monitored at m/z 391.10→149.30for shanzhiside, 573.40→365.05for genipin-1-gentiobioside, 447.30→225.15for geniposide.Results The regress equation of shanzhiside, genipin-1-gentiobioside and geniposide were Y=243.810 X-289.957, r=0.999 9;Y=137.125 X+2 092.76, r=0.999 6;Y=2 030.32 X+823 213, r=0.999 8in the range of 10.76-215.2ng/ml;516-4 128ng/ml;2 000-20 000ng/ml respectively.This validated method has good repeatability, precision, recovery and stability.The results meet the requirements by regulation.Conclusion This method shortened the analysis time and improved efficiency.It assayed the three iridoid glycosides in Gardenia jasminoides Ellis sensitively and precisely.This method can be used for the quality control of Gardenia jasminoides Ellis.

Genipin-Crosslinked Gelatin Scaffold in Tissue Engineering: A Systematic Review

@#Gelatin sering digunakan dalam pembuatan kerangka kejuruteraan tisu kerana ia mempunyai ciri-ciri biologi yang baik, termasuk mempercepatkan penyembuhan luka. Genipin, bahan semula jadi yang diperolehi dari tumbuhan Gardenia, terbukti berkesan dalam memperkukuhkan ciri-ciri fizikokimia gelatin. Ulasan sistematik ini melaporkan pengetahuan terkini mengenai pengunaan genipin sebagai agen penyilangan gelatin. Dua pangkalan data elektronik telah digunakan, iaitu Scopus dan MEDLINE melalui Ebscohost. Carian dilakukan untuk penerbitan antara Januari 1999 hingga Disember 2018 menggunakan kata kunci ‘gelatin’ dan ‘genipin’. Makalah berbahasa Inggeris, yang melaporkan kegunaan genipin untuk pembuatan span gelatin telah dipilih. Terdapat 830 makalah dijumpai melalui carian kata kunci di mana 14 makalah telah dipilih dan dibincangkan dalam ulasan sistematik ini. Dapatan kajian termasuk kepekatan, suhu penyilangan, dan cara pembuatan yang optima untuk genipin. Kepekatan genipin yang optima adalah 0.5% dan suhu penyilangan yang optima adalah 25˚C. Keputusan kajian ini menunjukkan jurang pengetahuan dalam pengunaan genipin sebagai agen penyilangan gelatin dan lebih kajian diperlukan untuk mengisi jurang ini. Kajian ini menyediakan tinjauan meluas berkenaan pengetahuan terkini mengenai penggunaan genipin sebagai agen penyilangan gelatin.

Genipin inhibits oxidative stress and apoptotic damage in high glucose-induced H9c2 cardiomyocytes / 中国病理生理杂志

AIM:To explore the effects of genipin (GEN) on high glucose (HG) -induced oxidative stress injury and apoptosis in H9c2 cardiomyocytes.METHODS:H9c2 cells were cultured in vitro and HG-induced injury model was established.H9c2 cells were divided into 4 groups:normal control (NC) group (glucose at 5.6 mmol/L) , HG group (glucose at 50 mmol/L) , NG+GEN group and HG+GEN group.The concentration of genipin was used at 10μmol/L.The viability of the H9c2 cells was measured by CCK-8 assay.The intracellular malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were determined by enzyme labeling and WST-1 methods, respectively.The activity of lactate dehydrogenase (LDH) in the cell culture supernatant was detected by microplate method.Fluorescent probe DCF was used to detect intracellular levels of reactive oxygen species (ROS).Nucleosome fragments was measured to evaluate cell apoptosis by ELISA.The intracellular mitochondrial membrane potential was detected by JC-1 method.The protein levels of Mn-SOD, cytochrome C (Cyt C) , Bax and cleaved caspase-3 were determined by Western blot.RESULTS:Compared with HG group, the cell viability in HG+GEN group was increased significantly (P<0.05) , the levels of MDA and LDH were decreased (P<0.05) , SOD activity was increased (P<0.05) , the levels of ROS and nucleosome fragments in HG+GEN group were decreased (P<0.05) , and the mitochondrial membranes potential was notably increased (P<0.05).Compared with NG group, the activation of Mn-SOD was decreased, but the protein levels of Cyt C, Bax and cleaved caspase-3 were increased in HG group (P<0.05).Compared with HG group, the activation of Mn-SOD was increased, and the protein levels of Cyt C, Bax and cleaved caspase-3 were decreased in HG+GEN group (P<0.05).CONCLUSION:Genipin protects HG-induced H9c2 cardiomyocytes against oxidative stress injury and apoptosis.

Efficiency and safety of genipin collagen crosslinking on rabbit cornea stroma model in vivo / 中华实验眼科杂志

Objective To evaluate the efficiency and safety of genipin collagen crosslinking (G-CXL) on rabbit corneas in vivo.Methods Forty healthy New Zealand white rabbits were randomly divided into 0.20％ G-CXL,0.25％ G-CXL,standard UVA-CXL and normal control group.And the right eyes were treated in different grouping.No procedures were performed in the normal control group.The corneal curvature (Km) and central corneal thickness (CCT) of right eyes were evaluated before,7 days and 14 days after crosslinking treatment.Cornea strips were harvested from the right eyes and tensile strain measurements were performed 7 days and 14 days after crosslinking treatment.The structure of corneal stroma was observed under light microscope (LM) and transmission electron microscope (TEM).Results No statistically significant differences in Km were observed among different groups or different timepoints (Fgroup =0.301,P=0.825;Ftime =1.287,P=0.284).Significant difference in CCTs was noticed among different time pionts (Ftime =3.786,P =0.029).Compared with preoperative,the CCTs of all the groups were significantly increased 7 days after crosslinking (all at P＜0.05).No significant difference in CCT was found among the groups (Fgroup =0.557,P=0.646).Seven days after crosslinking treatment,the Young's modulus at 10％ strain was (1 1.96±5.74),(21.24±6.77),(18.76±3.34) and (11.56±4.37) MPa in 0.20％ G-CXL group,0.25％ G-CXL group,UVA-CXL group and normal control group,respectively;the stress at 10％ strain was (0.68 ±0.24),(1.20 ± 0.25),(1.0l ± 0.30) and (0.69 ± 0.26) MPa,respectively;the Young's modulus and stress in 0.25％ G-CXL group was significantly increased when compared with those in 0.20％ G-CXL and normal control group (both at P＜0.05).No significant difference in Young's modulus and stress was observed between 0.25％ G-CXL group and UVA-CXL group (all at P＞0.05).Forteen days after crosslinking treatment,Young's modulus at 10％strain was (16.65±3.19),(19.12±2.39),(22.83 ±4.38) and (12.70±2.72)MPa in 0.20％ G-CXL group,0.25％ G-CXL group,UVA-CXL group and normal control group,respectively;stress at 10％ strain was (0.83 ±0.12),(0.97±0.04),(1.23±0.30) and (0.65±0.20) MPa,respectively;the Young's modulus and stress in UVA-CXL group was significantly increased,when compared with 0.20％ G-CXL group and normal control group (all at P＜0.05).Statistical significance of stress was observed between 0.25％ G-CXL group and UVA-CXL group (P =0.046).There is no significant difference in Young's modulus between 0.25％ G-CXL and UVA-CXL group (P =0.090).LM showed the reduction of keratocytes existed in superficial stroma of 0.20％ and 0.25 ％ G-CXL groups,while the reduction of keratocyte was found in anterior and intermediate stroma of UVA-CXL group.In 0.20％ and 0.25％ G-CXL groups,the ultrastructure of keratocytes was normal except vacuole in some keratocytes.Keratocytes apoptosis was noticed in UVA-CXL group and keratocytes was normal in deep stroma under TEM.Conclusions 0.25％ has a similar biomechanics effect when compared to UVA-CXL.Moreover,histological observation proves a better safety of G-CXL in comparison of UVA-CXL.

H NMR-based metabolomics approach to investigating the renal protective effects of Genipin in diabetic rats / 中国天然药物

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.

H NMR-based metabolomics approach to investigating the renal protective effects of Genipin in diabetic rats / 中国天然药物

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.

Influence of genipin in abilities of proliferation and invasion and apoptosis of gastric cancer SGC 7901 cells and their mechanisms in vitro / 吉林大学学报(医学版)

Objective: To investigate the regulatory effect of genipin (GP) on the gastric cancer SGC 7901 cells, and to clarify its mechanism. Methods: The SGC 7901 cells in logarithmic growth phase were selected and divided into control group and different concentrations (5. 0, 10. 0, and 20. 0 mg · L-1) of GP groups. MTT was used to detect the inhibitory rates of proliferation of SGC 7901 cells at different time points (24, 48, and 72 h). Transwell chamber cell invasion assay was used to detect the invasion ability of SGC 7901 cells in vitro. The expression levels of Bcl-2, Bax and caspase-3 proteins in the SGC 7901 cells were detected by Western blotting method. Results: The results of MTT assay showed that the inhibitory rates of proliferation of the cells in different concentrations of GP groups after treated with GP for different time were increased significantly compared with control group (P< 0. 01). The Transwell cell invasion assay results showed that compared with control group, the number of transmembrane cells in 10. 0 and 20. 0 mg · L-1 GP groups was decreased significantly after treated for 72 h (P< 0. 01). The results of Western blotting method showed that compared with control group, the expression level of Bcl-2 protein in 20. 0 mg · L-1 GP group was decreased after treated for 72 h (P<0. 01), and the expression levels of caspase-3 and Bax proteins were increased (P<0. 05). Conclusion; GP can inhibit the abilities of proliferation and invasion in vitro of SGC 7901 cells, and induce the apoptosis; its mechanism may be related to the regulation of Bcl-2, caspase-3, and Bax protein expressions.

Biophysical characteristics of genipin-crosslinked amniotic membrane bio-scaffold / 中华实验眼科杂志

Objective To investigate the characteristics and feasibility of genipin-crosslinked amniotic membrane(AM) as bio-scaffold.Methods Human umbilical cord mesenchymal stem cells (hUCMSCs) were isolated from fresh umbilical cord and cultured by adherent method.The expressions of PE-CD34,PE-CD45,PE-CD90,FITC-105 and FITC-Oct-4,the markers of hUCMSCs,were detected by flow cytometry.Alizarin red and oil red O staining were performed to identify the cells after adipogenesis and osteogenesis induction on the third-generation cells.Human AMs were treated at 37 ℃ and 45 ℃ by 0.5％ and 1％ genipin solution for 24,36 and 48 hours respectively,and the mechanical properties of AM in each group were measured and compared.The hUCMSCs were divided into only hUCMSCs culture group,fresh AM group,crosslinked AM group,gelatin group and crosslinked AM+gelatin group,and the cells were cultured in the corresponding medium.The content of hydroxyproline among the groups was detected with hydroxyproline kit,and proliferation of the cells (absorbance) was assayed by MTT method to evaluate the biological compatibility of crosslinked AM.Results The maximum tensile displacement of the crosslinked-AM by 0.5％ and 1％ genipin was (8.31±0.43)mm and (4.49±0.37)mm respectively,and those after crosslinked with 0.5％ genipin under the 37 ℃ and 45 ℃ for 24 hours was (9.89±1.09)mm and (5.39±0.59)mm,respectively,showing a significant difference between them (t =6.389,P＜0.05).The maximum tensile displacement of the crosslinked-AM was gradually decreased as the lapse of crosslinking time,and an insignificant difference was found among 24,36 and 48 hours after 0.5％ genipin treatment under the 37 ℃ (P＞0.05).The loading force of the crosslinked-AM was significantly higher in the 1％ genipin treated group than that in the 0.5％ genipin treated group (P＜0.05),and the loading force of the AM was significantly increased in 45 ℃,0.5％ genipin,24 hours crosslinked group compared with the 37 ℃,0.5％ genipin,24 hours crosslinked group (t =5.528,P＜0.05).The content of hydroxyproline in the AM was (1.28±0.36),(2.03 ±0.49) and (2.11 ±0.10) mg/g in the 1％ genipin crosslinked AM group,0.5％ genipin crosslinked AM group and fresh AM group,respectively,and the content of hydroxyproline in the AM in the 1％ genipin group was significantly lower than that in the 0.5％ genipin group in the fresh AM group (both at P＜0.05).The proliferative values of the hUCMSCs were significantly lower in the only hUCMSCs culture group,fresh AM group and gelatin group were significantly reduced in comparison with the crosslinked AM group and crosslinked AM+gelatin group (all at P＜0.05).There was no significant difference in the proliferative values of the hUCMSCs between crosslinked AM group and crosslinked AM+gelatin group (P＞0.05).Conclusions Different crosslinked temprature,crosslinking period and concentration of genipin impact the mechanical properties of AM.Crosslinked AM with genipin is feasible as a carrier scaffold of artificial cornea because of less tissue toxicity and better plasticity.

Quantitative study of nine main components in Yueju Tablets by HPLC coupled with wavelength switching and gradient elution method / 中草药

Objective: To establish HPLC coupled with wavelength switching and gradient elution method (HPLC-DVD) for simultaneous determination of nine main components (α-cyperone, genipin-1-β-D-gentiobioside, geniposide, crocin I, senkyunolide H, senkyunolide I, senkyunolide A, ligustilide, and atractylodin) in Yueju Tablets. Methods: The chromatographic separation was achieved on Zorbax Eclipse Plus C18 (250 mm × 4.6 mm, 5 μm) column with methanol-acetonitrile (2∶1) (A)-0.2% glacial acetic acid solution (B) as mobile phases for gradient elution, at the flow rate of 0.9 mL/min; The detection wavelength was set at 240 nm for α-cyperone, genipin-1-β-D-gentiobioside and geniposide, 440 nm for crocin I, 280 nm for senkyunolide H, senkyunolide I, senkyunolide A and ligustilide, and 340 nm for atractylodin. The volume of sample injection was 10 μL. Results: The nine active components were well separated and showed good linearity, such as α-cyperone 2.58—51.60 μg/mL (r = 0.999 4), genipin-1-β-D- gentiobioside 11.99—239.80 μg/mL (r = 0.999 9), geniposide 17.96—359.20 μg/mL (r = 0.999 6), crocin-I 3.98—79.60 μg/mL (r = 0.999 7), senkyunolide H 2.82—56.40 μg/mL (r = 0.999 9), senkyunolide I 2.38—47.60 μg/mL (r = 0.999 9), senkyunolide A 6.04—120.80 μg/mL(r = 0.999 5), ligustilide 7.98—159.60 μg/mL (r = 0.999 3), and atractylodin 6.51—130.20 μg/mL (r = 0.999 2). The precision was good, and RSD was not more than 1.22%. The repeatability was good, and RSD was not more than 1.75%. The stability was good in 18 h, and RSD was not more than 1.37%. The average recoveries and corresponding RSD values were 97.64% (0.98%), 99.09% (1.46%), 100.11% (1.03%), 97.87% (0.80%), 98.59% (1.19%), 96.89% (1.34%), 99.38% (0.58%), 98.50% (1.22%), and 99.71% (0.85%), respectively. The contents of 10 batches of α-cyperone, genipin-1-β-D-gentiobioside, geniposide, crocin I, senkyunolide, H, senkyunolide I, senkyunolide A, ligustilide and atractylodin were 0.282—0.344, 2.099—2.445, 3.628—4.225, 0.758—0.913, 0.241—0.286, 0.217—0.266, 1.077—1.291, 1.386—1.623, and 1.137—1.434 mg/tablet. Conclusion: HPLC coupled with wavelength switching and gradient elution method has been established for simultaneous determination of nine components in Yueju Tablets. The method is simple, quick, accurate, and it can be used for content determination and quality control of Yueju Tablets.

A pilot study of genipin cross-linking effects on bullous keratopathy in rabbits / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To evaluate the effects of genipin cross-linking on bullous keratopathy in rabbits. <p>METHODS: Nine female New Zealand white rabbits with bullous keratopathy were used as an experimental model. They were randomized into three groups. Corneas in Group A(treatment group, <i>n</i>=3)were immersed in 0.25% genipin at 24℃ for 40min; those in Group B(control group, <i>n</i>=3)were immersed in 0.9% sodium chloride solution at 24℃ for 40min; and those in Group C(blank control group, <i>n</i>=3)received no treatment. Follow-up examinations were performed within 2wk after treatment, including slit-lamp microscopy, central corneal thickness(CCT), evaluations of body weight and stress responses, histopathological analyses, and terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)for detecting stromal cell apoptosis. <p>RESULTS: Compared to Groups B and C, remission of corneal edema, corneal healing, disappearance of epithelial bullae, a significant decrease in CCT(<i>P</i><0.05), and a significant increase in body weight(<i>P<</i>0.05)were found in Group A. Animals in Group A became more active and showed less aggression and violent resistance behavior. More regular and dense arrangement of collagen fibers in the corneal stroma and formation of blue strips of cross-linking products were observed in Group A. Cell apoptosis occasionally occurred in the corneal stroma of Group A, while no cell apoptosis was observed in Groups B and C. <p>CONCLUSION: Genipin cross-linking treatment for bullous keratopathy in rabbits results in remission of corneal edema and relief of pain. We hypothesize that genipin cross-linking strengthens collagen fibers in corneal stroma to avoid the formation of corneal edema and bullae.

Investigation on Endogenous Metabolites in Pancreas of Diabetic Rats after Treatment by Genipin through 1H-NMR-based Metabolomic Profiles / 中草药·英文版

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.