Effect of Geniposide on Liver Injury and Bile Acid Profile in Serum, Liver and Excrement of Rats / 中国实验方剂学杂志

Objective:To investigate the hepatotoxicity of different doses of geniposide on the liver of rats and the effects on bile acid profile in serum， liver tissue and feces. Method:The 60 Sprague Dawley rats， half male and half female， were randomly divided into 5 groups according to body weight： blank group and four different doses （50， 100， 200， 400 mg·kg^-1） geniposide groups， 12 rats in each group. The rats were treated by gavage once a day for 7 consecutive days， and the serum， liver and cecal contents were collected on the 8^th day of treatment. The activities of alanine aminotransferase （ALT）， aspartate aminotransferase （AST） and alkaline phosphatase （ALP）， the contents of albumin （ALB）， total bilirubin （TBIL）， total bile acid （TBA）， creatinine （Crea） and carbamide （Urea） were detected in each group. The sections of liver tissue were stained with hematoxylin-eosin（HE）， and the protein expressions of cytokeratin 7（CK7） and cytokeratin 19（CK19） were detected by immunohistochemistry. The protein expressions of CK7 and CK19 in the liver tissue were detected by Western blot. And the mRNA expressions of cholesterol 7<italic>α</italic>-hydroxylase （CYP7A1）， cholesterol 27<italic>α</italic>-hydroxylase （ CYP27A1） and cholesterol 12<italic>α</italic>-hydroxylase （CYP8B1） were detected by real-time PCR. The contents of 18 kinds of bile acids in serum， liver and cecal contents were determined by ultra-performance liquid chromatography-mass spectrometry（UPLC-MS）. Result:Compared with the control group， TBIL level in each dose of geniposide group was increasesd significantly （<italic>P</italic><0.01）. ALT， AST activity and TBA content in 400 mg·kg^-1 geniposide group were increased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）. HE staining showed that， compared with control group， there was bile duct reaction in the portal area and inflammatory cells infiltrate around bile duct in 200 mg·kg^-1 and 400 mg·kg^-1 geniposide groups， especially 400 mg·kg^-1. The expressions of CK7 and CK19 in liver tissue of 400 mg·kg^-1 geniposide group were significantly higher than those in the control group （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the control group， the contents of glycoursodeoxycholic acid （GUDCA） and glycohyodeoxycholic acid （GHDCA） in liver tissue of 400 mg·kg^-1 geniposide group decreased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）， the contents of sodium taurochenodeoxycholate （TCDCA）， hyodeoxycholic acid （HDCA）， cholic acid （CA） and chenodeoxycholic acid （CDCA） in liver tissue increased significantly （<italic>P</italic><0.01）， the contents of glycocholic acid hydrate （GCA）， glycochenodeoxycholic acid （GCDCA）， glycodeoxycholic acid hydrate （GDCA）， glycocholic acid （GLCA）， tauroursodeoxycholic acid （TUDCA）， GUDCA， GHDCA， ursodeoxycholic （UDCA） and taurolithocholic acid （TLCA） decreased， the proportions of TCDCA， HDCA， CA， CDCA and deoxycholic acid （DCA） in liver tissue increased， the contents of GHDCA and lithocholic acid （LCA） in serum decreased significantly （<italic>P</italic><0.01）， while sodium taurohyodeoxycholate hydrate （THDCA）， taurocholic acid （TCA）， GCA， TCDCA， UDCA， CA， CDCA， DCA in serum decreased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）. The contents of CA， UDCA， CA， CDCA and DCA increased significantly （<italic>P</italic><0.05）， the ratio of CA/DCA increased significantly （<italic>P</italic><0.05）， and the ratio of CA and CDCA increased by 19.60% and 4.63%， respectively； Compared with the control group， the contents of all bile acids in cecal contents of 400 mg·kg^-1 were decreased， and the contents of GCA， UDCA， HDCA， GCDCA， GDCA， TLCA， GLCA， CDCA， DCA and LCA were decreased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）. In addition， real-time PCR results showed that the mRNA expressions of CYP7A1， CYP27A1 in the 400 mg·kg^-1 geniposide group were significantly higher than those in the control group （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:The 400 mg·kg^-1geniposide can cause obvious hepatotoxicity in rats， and the bile acid profile in liver， serum and excrement changes significantly， and the changes of the each bile acid in liver， serum and feces are different. However， the causal relationship between the gardenoside-induced liver injury and the changes in bile acid profile are<italic> </italic>not clear. It needs to be further studied.

Effect of Xiayuxue Tang with Different Processing Method on CCl4-induced Hepatic Fibrosis in Rats / 中国实验方剂学杂志

Objective::To compare the anti-hepatic fibrosis effect of ethanol extract of honey Xiayuxue Tang (H-Xiayuxue Tang)and prepared slices of crude drug-Xiayuxue Tang (C-Xiayuxue Tang). Method::Totally 40 wistar rats were randomlydivided into normal group, model group, H-Xiayuxue Tang group, C-Xiayuxue Tang group, and Fuzheng Huayu group. Liver fibrosis model was induced by hypodermic injection of 50% carbon tetrachloride for 9 weeks. From the seventh week, the normal group and the model group were given 0.3% sodium carboxymethyl cellulose by gavage. The other three groups were given H-Xiayuxue Tang(0.58 g·kg-1), C-Xiayuxue Tang (0.43 g·kg-1), Fuzheng Huayu (2 g·kg-1), respectively. At the end of the ninth week, liver tissues and serum samples were collected for follow-up studies. The pathological changes and fibrosis degree were observed by HE and Sirius red staining, hydroxyproline content in liver tissues was detected by alkaline hydrolysis, serum liver function was determined by automatic analysis, the expressions of collagen type I (COL-I) and α-smooth muscle actin (α-SMA) in liver tissues were detected by Real-time PCR. The expressions of α-SMA in liver tissues was detected by Western blot and Immunohistochemistry, and serum inflammatory factors and oxidantion indexes were determined by ELISA, respectively. Result::Compared with control group, the severity of liver fibrosis was observed, the area ratio of Sirius red positive staining, hydroxyproline (Hyp) content, the protein and mRNA expressions of α-SMA, COL-I in the liver tissue were significantly increased in the model group (P<0.01). And the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TBil), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-17A (IL-17A) and malondialdehyde (MDA) in the serum of the rats were markedly increased (P<0.01). The levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were apparently decreased (P<0.01). Compared with model group, the degree of liver fibrosis was significantly improved, the area ratio of Sirius red positive staining, the content of Hyp, the protein and mRNA expression of α-SMA and COL-I, and the serum levels of AST, ALT, TBil, TNF-α, IL-1β, IL-17A and MDA were significantly decreased (P<0.05, P<0.01). The serum contents of SOD and GSH-Px were markedly increased (P<0.05, P<0.01), respectively, by H-Xiayuxue Tang or C-Xiayuxue Tang. Among them, the improvement of liver histopathological changes was more obvious in the H-Xiayuxue Tang group than in the C-Xiayuxue Tang group. Meanwhile, the percentage of Sirius red positive staining area, the content of hydroxyproline, the expressions of COL-I and α-SMA mRNA and protein in liver tissue were significantly decreased compared with the C-Xiayuxue Tang group(P<0.05, P<0.01). In addition, there was no significant difference in serum liver function, inflammatory factors and oxidantion related indicators between groups, but the improvement trend of H-Xiayuxue Tang was more obvious. Conclusion::H-Xiayuxue Tang and C-Xiayuxue Tang have a better effect in hepatic fibrosis, especially H-Xiayuxue Tang, suggesting that honey can increase the anti-hepatic fibrosis effect of Xiayuxue Tang.

A Novel Screening Method for Isolating High Cholesterol Oxidase Producing Strain / 微生物学通报

The cholesterol oxidase producing strain Brevibacterium sp.DGCDC-82 was treated with NTG (1 mg/mL)under ultrasonicztion(200 W,50 kHz).A red mutant named Brevibacterium sp.DGCCN-25 showed higher and stable production of cholesterol oxidase was obtained.The enzyme activity was increased by 140%,it is 1.24 U/mL.Then dealed with DGCCN-25 using the same method,two revertants were obtained,one was white and the other was rose pink.The enzyme activity of two revertants was obvious decrease,they are 0.17 U/mL and 0.69 U/mL.The results showed the positive correlation between COD acticity and red pigment producing by Brevibacterium sp..The relativity model can be used as a method of screening for mutation and directed evolution.