异烟肼、利福平对小鼠胆汁酸转运体Ntcp、Bsep的表达影响研究

发布时间：2018-05-12 15:56

本文选题：异烟肼 + 利福平　；参考：《中南大学》2014年硕士论文

【摘要】：目的：研究异烟肼(INH)与利福平(RFP)单用及合用致小鼠肝损伤模型中对肝细胞上胆汁酸有关的转运体Ntcp、Bsep表达的影响。探讨利福平与异烟肼合用引起肝损伤加重是否与这些转运体表达变化有关。方法：选用SPF级56只成年雌性ICR小鼠(ICR mouse)(29.8±1.4g),按照完全随机分组原则分为7组,实验组分别给与50mg/kg INH,100mg/kg RFP,50mg/kg+100mg/kg INH+RFP,100mg/kg INH,200mg/kg RFP,100mg/kg+200mg/kg INH+RFP,对照组给予空白溶剂,连续给药2周,第15天将小鼠以水合氯醛麻醉,摘除眼球取血,离心取上清,用于肝功能生化指标测定,每组每只留取肝脏组织分别于10%福尔马林固定用于病理切片、HE染色；4%多聚甲醛固定用于目标蛋白免疫组化测定；Western blot测定目标蛋白Ntcp及Bsep的表达。结果：高剂量INH+RFP能够引起生化指标ALT、AST、TBil、DBil、ALP、TBA、MDA显著变化,肝脏形态及病理结果显示有肝损伤,低剂量INH、RFP、INH+RFP则变化较小。免疫组化及Western blot结果示对照组和各实验组Ntcp及Bsep均有表达,单用组及合用组较正常对照组相比,Ntcp蛋白表达均下调(P0.01),H-INH+RFP较L-INH+RFP组Bsep表达明显下调(P0.01)。Ntcp、Bsep蛋白的表达与血清中胆汁酸的分泌量呈负相关,相关系数分别为-0.320,-0.391,差异有统计学意义(P0.05)。结论：高剂量INH、RFP单用及合用均造成小鼠肝损伤,INH单用及与RFP合用均能有下调胆汁酸有关转运体Ntcp及Bsep蛋白的表达,高剂量时对转运体Bsep的表达影响更为明显,INH单用及与RFP合用诱导小鼠肝损伤模型的血清生化指标与组织病理学的变化明显滞后于转运体的显著变化,INH、RFP引起肝损伤病理状态可能是转运体与内源性物质(胆红素、胆汁酸等)改变共同作用的结果。图13幅,表4个,参考文献89篇。
[Abstract]:Aim: to study the effect of isoniazid (INH) and rifampicin (RFP) alone or in combination on the expression of bile acid-related transporter NtcpnBsep in hepatocytes. To investigate whether rifampicin combined with isoniazid can aggravate liver injury and change the expression of these transporters. Methods: 56 adult female ICR mice of SPF grade were randomly divided into 7 groups according to the principle of complete randomized grouping. The experimental group was given 100 mg / kg 100mg/kg INH 100mg/kg INH 100 mg / kg 100mg/kg INH rpm 100 mg / kg 100mg/kg INH 100 mg / kg 200mg/kg INH RFP, respectively. The control group was given blank solvent for 2 weeks. On the 15th day, the mice were anesthetized with chloral hydrate, blood was removed from eyeball, supernatant was centrifuged, and used to determine the biochemical index of liver function. Each liver was fixed with 10% formalin for HE staining and 4% paraformaldehyde fixation to determine the expression of Ntcp and Bsep by immunohistochemistry. Results: high dose of INH RFP could cause significant changes of MDA in the biochemical index of ALTAST TBildlb TBAM, liver morphology and pathological results showed liver injury, while low dose of INHG RFPINH RFP showed little change. The results of immunohistochemistry and Western blot showed that Ntcp and Bsep were expressed in the control group and each experimental group. Compared with the control group, the expression of ntcp protein decreased significantly in the single group and the combination group compared with the control group. The expression of Bsep in the single group and the combination group was significantly lower than that in the group of L-INH RFP. There was a negative correlation between the expression of the protein and the secretion of bile acid in serum, the correlation coefficient was -0.320-0.391.The difference was significant (P 0.05). Conclusion: the expression of Ntcp and Bsep protein of bile acid-related transporter can be down-regulated by both high-dose INHP-RFP alone and its combination with RFP in mice liver injury. The effect of high dose on the expression of transporter Bsep was more obvious. The changes of serum biochemical indexes and histopathology in mice model of liver injury induced by INH alone and combined with RFP were obviously lagging behind the significant changes of transporter. The pathological state may be transporters and endogenous substances (bilirubin, bilirubin, Bile acids, etc.) change the result of a common action. 13 figures, 4 tables, 89 references.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965

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