The aim of this paper was to screen out relevant genes of geniposide-induced hepatotoxicity based on genomics,in order to provide a scientific basis for the non-clinical evaluation of drugs containing Gardeniae Fructus and geniposide. Fifty-five SD rats were randomly divided into normal control group,24 h group and 72 h group. The changes of appearance,behavior and weight of rats were observed after administration by gavage for 3 days. The activities of ALT and AST were detected. Molecular mechanism of geniposideinduced hepatotoxicity was investigated by Affymetrix miRNA 4. 0 and Affymetrix Rat Gene 2. 0 to examine the gene expression levels in Sprague-Dawley rat livers at 24 h and 72 h after administration of overdose-geniposide( 300 mg·kg-1 daily),and then verified by Realtime quantitative PCR. Compared with the normal control group,the activities of ALT and AST were markedly increased. In addition,experimental results indicated that 324 genes were differentially expressed,among which 259 were up-regulated and 65 down-regulated.Nine candidate genes were verified by qRT-PCR,including Bcl2,Il1 b,Tpm3,MMP2,Col1α1,Ifit1,Aldob,Nr0 b2,Cyp2 c23. And Bcl2,Col1α1,Aldob,Nr0 b2 and Cyp2 c23 were found to be correlated with geniposide-induced hepatotoxicity. This study provides an important clue for mechanism of geniposide-induced hepatotoxicity.
Animals ; Biomarkers/metabolism* ; Chemical and Drug Induced Liver Injury ; Genomics ; Iridoids/toxicity* ; Liver/metabolism* ; Rats ; Rats, Sprague-Dawley

Investigated the changes of crosslinking index, swelling ratio, degradation rate and cytotoxicity of genipin crosslinked gelatin accompany with crosslinking time. 1% genipin crosslinked gelatin were divided into 7 groups by crosslinking time: 10 min group, 30 min group, 1 h group, 2 h group, 12 h group, 24 h group, 72 h group. The results proved that genipin could crosslink gelatin effectively. Accompany with increasing of crosslinking time, crosslinking index increased, and swelling ratio, degradation rate decreased. In 10 min group, crosslinking index was low(26.7%), swelling ratio was high, (265%), completely degraded within 1 week. This indicated that biomaterials of 10 min group was instable and degraded easily. Compared with 10 min group, biomaterials of 30 min group changed significantly with crosslinking index(45.7%), swelling ratio (206%) and degration rate (completely degraded between 4 weeks and 8 weeks). This indicated that genipin could change the properties of gelatin within 30 min. Biomaterials after 30 min, crosslinking index increased, and swelling ratio, degradation rate decreased gradually accompanied with increasing of crosslinking time. Biomaterials of 72 h, crosslinking index was 73.1%, swelling ratio was 152%, and degradated 18.9% after 12 weeks. RGR (relative cell growth rate) of every group measured by MTT assay changed between 87.9% and 105.4%, indicated that the cytotoxicity of genipin crosslinked gelatin was very low.
Animals ; Biodegradation, Environmental ; Cell Survival ; drug effects ; Cricetinae ; Cricetulus ; Cross-Linking Reagents ; chemistry ; toxicity ; Gelatin ; chemistry ; Iridoid Glycosides ; Iridoids ; chemistry ; toxicity ; Time Factors

OBJECTIVETo purify the extract of fructus gardeniae for injection by macroreticular resins in purification process of traditional Chinese medicine (TCM) Injections.

METHODUsing fructus gardeniae as sample, on base of obtaining the extract by employing macroreticular resin, quality evaluation and rationality of purification methods had been studied by the quantitative analysis of active ingredients and the characteristics of micromeritics, safety and stability of the extract.

RESULTThe experiment showed the extract of fructus gardeniae for injection had been produced successfully by macroreticular resin.

CONCLUSIONUsing macroreticular resins is a promising purification way of TCM injections, whereas a more consummate method of quality evaluation must be established to ensure safety, efficiency and stability of the preparation in the process.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; toxicity ; Fruit ; chemistry ; Gardenia ; chemistry ; Injections ; Iridoids ; isolation & purification ; Mice ; Plants, Medicinal ; chemistry ; Pyrans ; isolation & purification ; Quality Control ; Resins, Synthetic ; Technology, Pharmaceutical ; methods

OBJECTIVETo study the hepatotoxicity effects in rats with different extract of Fructus Gardeniae.

METHODObserve the change of appearance, behavior and weight of rats through oral gavage daily for 3 d. Weigh the liver and calculate the liver index. Detect the ALT, AST and TBIL. Observe the liver tissue by optical microscope.

RESULTThe weight and index of liver were increased by 3.08 g x kg(-1) aqueous extract, 1.62 g x kg(-1) alcoholic extract and 0.28 g x kg(-1) geniposide, compared to those of the blank group (P < 0.005, P < 0.001) and the activities of ALT, AST and the content of TBIL were also increased, compared to those of the blank group (P < 0.05, P < 0.001). The liver cells were obviously swell, necrotic and changed with inflammatory infiltrate.

CONCLUSIONAqueous extract, alcoholic extract and geniposide displayed hepatotoxicity, and the geniposide which was the main substance of the Fructus Gardeniae might be mainly responsible for the hepatotoxicity.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Bilirubin ; blood ; Drugs, Chinese Herbal ; isolation & purification ; toxicity ; Female ; Fruit ; chemistry ; Gardenia ; chemistry ; Iridoids ; isolation & purification ; toxicity ; Liver ; pathology ; Male ; Organ Size ; drug effects ; Plants, Medicinal ; chemistry ; Pyrans ; isolation & purification ; toxicity ; Random Allocation ; Rats ; Rats, Sprague-Dawley

BACKGROUNDUncoupling protein (UCP) 2 is related to the dysfunction of beta cells induced by fatty acids. However, whether UCP2 has similar effects on alpha cell is still not clear. This study aimed to investigate the effects of UCP2 and its possible mechanisms in lipotoxicity-induced dysfunction of pancreatic alpha cells.

METHODSThe alpha TC1-6 cells were used in this study to evaluate the effects of palmitate and/or UCP2 inhibit factors on the glucagon secretory function, glucagon content, the glucagon mRNA level and the nitrotyrosine level in the supernatant. Meantime, the expression levels of UCP2 and peroxisome proliferator-activated receptor-γ coactivator-1 alpha (PGC-1 alpha) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. Furthermore, the possible relationship between UCP2 and insulin signal transduction pathway was analyzed.

RESULTSPalmitate stimulated alpha cell glucagon secretion and the expression of UCP2 and PGC-1 alpha, which could be partially decreased by the inhibition of UCP2. Palmitate increased nitrotyrosine level and suppressed insulin signal transduction pathway in alpha cells. Inhibition of UCP2 influenced the effects of free fatty acid on alpha cells and may relate to glucagon secretion.

CONCLUSIONUCP2 played an important role on alpha cell dysfunction induced by free fatty acid in vitro, which may be related to its effects on oxidative stress and insulin signal transduction pathway.

Animals ; Cells, Cultured ; Glucagon ; secretion ; Glucagon-Secreting Cells ; drug effects ; physiology ; Insulin ; pharmacology ; Insulin Receptor Substrate Proteins ; metabolism ; Ion Channels ; genetics ; physiology ; Iridoid Glycosides ; pharmacology ; Iridoids ; Mice ; Mitochondrial Proteins ; genetics ; physiology ; Oxidative Stress ; Palmitic Acid ; toxicity ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Phosphorylation ; RNA, Messenger ; analysis ; Signal Transduction ; Trans-Activators ; genetics ; physiology ; Transcription Factors ; Tyrosine ; analogs & derivatives ; metabolism ; Uncoupling Protein 2

BACKGROUNDA number of studies have shown that oxidative stress and mitochondrial involvement are major triggering factors in the development of neurodegenerative diseases. Cobalt chloride (CoCl(2))-induced cell death in PC12 cells may serve a simple and convenient in vitro model of hypoxia-induced neuronal cytotoxicity. To explore the effect of geniposide on CoCl(2) which induced cytotoxicity and mitochondrial function in rat pheochromocytoma PC12 cells, we analyzed the influence of geniposide on the expression of apoptosis-related proteins.

METHODSPC12 cells and RNAi PC12 cells were treated with 0, 12.5, 25, 50, 100 micromol/L geniposide for 12 hours and then exposure to 400 micromol/L CoCl(2) for 12 hours. Cell viability, cell morphology, and expression of Bcl-2, Bax, P53 and caspase-9 were determined using Western blotting.

RESULTSPretreatment with geniposide markedly improved the cells viability and morphology, decreased the expression of Bax, P53 and caspase-9, and increased the expression of Bcl-2 in PC12 cells challenged by CoCl(2)2. However, in the RNAi PC12 cells, geniposide had no significant effect on the expression of these proteins.

CONCLUSIONGeniposide protects PC12 cells from CoCl(2) involved in mitochondrial mediated apoptosis, and GLP-1R might play a critical role in the neuroprotection of geniposide in PC12 cells.

Animals ; Apoptosis ; drug effects ; Cobalt ; toxicity ; Glucagon-Like Peptide-1 Receptor ; Iridoids ; pharmacology ; Mitochondria ; physiology ; Neuroprotective Agents ; pharmacology ; PC12 Cells ; Proto-Oncogene Proteins c-bcl-2 ; physiology ; Rats ; Receptors, Glucagon ; drug effects ; physiology ; Signal Transduction ; bcl-2-Associated X Protein ; physiology