Altered integrity of hepatocyte tight junctions in rats with triptolide-induced cholestasis / 中国天然药物

Triptolide (TP), an active component of Tripterygium wilfordiiHook. f. (TWHF), has been widely used for centuries as a traditional Chinese medicine. However, the clinical application of TP has been restricted due to multitarget toxicity, such as hepatotoxicity. In this study, 28 days of oral TP administration (100, 200, or 400 μg·kg

Study on potential hepatotoxicity of main monomers of Polygonum multiflorum based on liver micro-tissue / 中国中药杂志

In this study, we aimed to establish a rat liver micro-tissue evaluation system to evaluate the hepatotoxicity of the main monomers in Polygonum multiflorum. Rat primary hepatocytes were isolated and purified by two-step in situ perfusion method to prepare hepatic parenchymal cells. The ultra-low adsorption plate and the inverted model were used to establish an in vitro hepatotoxicity evaluation system. After the system was established, the main monomer components(monanthone with emodin type, rhein, emodin, emodin-8-O-β-D-glucopyranoside, physcion) of P. multiflorum were selected for in vitro hepatotoxicity evaluation. This study showed that the primary cells of the liver can form liver micro-tissues in the low adsorption plate method and the mold perfusion method, with good liver structure and function, which can be used to evaluate the hepatotoxicity of the drug to be tested after long-term administration. The five monomers to be tested in P. multiflorum can significantly affect the proliferation of primary liver micro-tissues in rats in a dose-and time-dependent manner. The hepatotoxic effects were as follows: monanthone with emodin type > rhein > emodin > emodin-8-O-β-D-glucopyranoside > physcion. The results suggested that the emodin-type monoterpene and rhein might be the potential hepatotoxic components, while the metabolites of emodin-8-O-β-D-glucoside and emodin methyl ether showed more toxic risks. The rat primary hepatocyte micro-tissue model system established in this experiment could be used to achieve long-term drug administration in vitro, which was consistent with the clinical features of liver injury caused by long-term use of P. multiflorum. The experimental results provided important information and reference on the clinical application and toxic component of P. multiflorum.

Effects of isorhamnetin on human liver microsomes CYPs and rat primary hepatocytes / 药物评价研究

Objective To study the inhibitory effects ofisorhamnetin on six kinds of CYPs of liver in vitro,and the toxic effect on rat hepatocytes Methods This report uses warm incubation of human liver microsomes in vitro to investigate the inhibition of isorhamnetin on 6 kinds of CYPs (CYP2C19,CYP2D6,CYP3A4,CYP2E1,CYP1A2 and CYP2C9),and using HPLC-MS/MS to detect product of metabolism as well as analysing of the pathways of metabolic.At the same time,using rat primary hepatocytes which has low CYPs activity in vitro to explore whether the use of isorhamnetin will cause effects on the ALT,AST and LDH of hepatocytes.Results Isorhamnetin has inhibition effects on CYP2E1 and CYP1A2,the inhibition rate were 59.48％ and 39.91％,respectively.Methylated metabolite is produced after incubating of isorhamnetin and HLMs.The isorhmnetin becomes high polarity and water solubility metabolite 3,3',4',5,7-hydroxyflavone.Isorhamnetin of 30,100 and 300 μmol/L cause a significant rise of ALT and LDH in primary cultured rat hepatocytes cultured (P ＜ 0.01).isorharnnetin of 100 μmol/L cause a rise of AST in primary cultured rat hepatocytes cultured (P ＜ 0.05) and 300 μmol/L cause a significant rise (P ＜ 0.01).It was a dose-dependent manner.Conclusion Isorhamnetin in vitro mainly metabolized by HLMs,and at the same time have a certain inhibitory effect on CYP2E1 and CYP1A2,which may cause the drugs which are metabolized by CYP2E1 and CYP1A2 in vivo accumulation that lead to a series of drug interactions.The results also indicate that heavy use of isorhamnetin cause some adverse effects on hepatocytes,and it was a dose-dependent manner.Individuals need to pay attention to the dose ofisorhamnetin and the potential drug interactions.

Effects of Aplysin on ethanol-induced oxidative damage in rat primary hepatocytes / 中国药理学通报

Aim To investigate the protective effects of Aplysin on ethanol-induced oxidative damage in rat pri-mary hepatocytes. Methods Rat primary hepatocytes were obtained via the portal vein collagenaseⅣin situ perfusion technique followed by a Percoll density gradi-ent centrifuge. MTT test was used to determine the op-timum dose of Aplysin and ethanol, and detect the cell vitality in primary hepatocytes. Supernatants of primary hepatocytes were harvested to measure AST and LDH level, and the SOD, GSH-PX activities and MDA con-tent in primary hepatocytes were observed. Flow cy-tometry was used to detect the cell apoptosis rate. DNA damage in primary hepatocytes was detected by single-cell gel electrophoresis assay. The level of mitochon-drial membrane potential in primary hepatocytes was tested by fluorogenic probe JC-1 . The CYP2 E1 activity in primary hepatocytes was detected by colorimetry. The proteins of CYP2 E1 were detected by Western blot. Results 300 mmol·L-1 dose of ethanol and 30 mg·L-1 dose of Aplysin were the optimal dosages and were used in the subsequent experiments. Hepatocyte vitality was significantly increased in Aplysin group compared to that in ethanol group, and Aplysin inhibi-ted the release of AST and LDH(P<0. 05). For Apl-ysin treatment group, the activities of hepatocyte SOD and GSH were significantly increased, and MDA was markedly lowered as compared with those in ethanol group( P <0. 05 ) . Aplysin could alleviate hepatocyte apoptosis significantly, and hepatocyte DNA damage rates of Ⅱ ~Ⅲ level and Ⅳ level were significantly lowered in Aplysin treatment group as compared with those in ethanol group, and Aplysin had evident im-provement in alcohol induced mitochondria damage of hepatocyte. Primary hepatocyte activities and protein expression of CYP2 E1 were markedly lowered in Aply-sin treatment group as compared with those in ethanol group(P<0. 05). Conclusion Aplysin has protective effects on liver oxidative damage induced by alcohol of primary cultured rat hepatocytes by blocking CYP2 E1 activation, relieving oxidative stress, and sharpening the oxidation resistance ability.

Study on the infection of primary tupaia hepatocytes with hepatitis B virus in vitro / 中华微生物学和免疫学杂志

Objective To provide a better cell model of closely nature infectious state for further research of hepatitis B virus(HBV). Methods Primary tupaia hepatocytes were isolated by the two-step perfusion method. The hepatocytes were then infected with purified serum from patients with hepatitis B. DNA and RNA isolated from the hepatocytes were detected with Southern blot and Northern blot. HBsAg in supernatant was tested by immunohistochemical method. Results cccDNA, pgRNA and sgRNA could be detected by Southern blot and Northem blot, and strong signals could be seen from day 7 to day 14 post-in-fection. The S/CO value of HBsAg in supernatant decreased from day 1 to day 5 and then increased after 5 day. Conclusion Primary tupaia hepatocytes are competent for infection with HBV. HBV can stably repli-cate and express in HBV-infected tupaia hepatocytes.

Mutagenicity of Organic Extract From Drinking Water in a Certain City / 环境与健康杂志

Objective To have a knowledge of the mutagenicity of contaminates in drinking water in a certain city. Methods Mutagenic activity of organic extracts of source water chloridized water in water plant and tap water from the city were detected with single cell gel-electrophoresis mouse primary hepatocytes in culture were used and Ames test. Results For Ames test the organic extract in 3 liters chloridized water in water plant and tap water samples produced positive result and that in 6 liters source water sample still gave a negative result. For single cell gel-electrophoresis test the organic extract in 0.1 liter chloridized water in water plant tap water and 0.5 liter source water produced positive result. Conclusion The sensitivity of single cell gel-electrophoresis with primary hepatocytes in detection of genotoxicity of cloridized water and source water is much higher than Ames test.