目的 观察苦参碱(MT)对α-萘异硫氰酸酯(ANIT)诱导的胆汁淤积性大鼠的预防性保护作用,并探索其可能的作用机制。方法 30只SD大鼠随机分为正常对照组,模型组,MT低剂量(5 mgkg-1),MT高剂量组(10 mgkg-1)和阳性药对照组(熊去氧胆酸,UDCA 100 mgkg-1),每组6只,连续给药7 d。除正常对照组外,其余各组于实验第5天给予60 mgkg-1 ANIT;末次给药后禁食24 h取血检测总胆红素(TBIL)、天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)、碱性磷酸酶(ALP),并行HE染色,免疫组化(IHC)分析CYP3A4表达,蛋白质免疫印迹法(Western blot,WB)检测大鼠肝脏中CYP3A4和PXR蛋白的表达。结果 MT组较模型组显著降低TBIL、AST、ALT和ALP,呈剂量依赖性;IHC法显示,MT高剂量组肝组织中CYP3A4表达较模型组显著上调(P＜0.05);WB法结果显示,MT组CYP3A4、PXR蛋白表达较模型组显著上调(P＜0.05),UDCA组CYP3A4显著上调(P＜0.05)。结论 MT能改善胆汁淤积性肝损伤,可能的机制在于MT上调CYP3A4表达,而CYP3A4的表达上调可能与PXR诱导有关。
Abstract
OBJECTIVE To investigate the preventive protective effect of matrine(MT) on α-naphthl isocyanate(ANIT)-induced cholestasis in rat, and to explore its possible mechanism. METHODS Thirty SD rats were randomly divided into five groups with six rats in each group: normal control group, model group, low dose matrine(5 mgkg-1), high dose matrine group(10 mgkg-1) and positive control group(ursodeoxycholic acid 100 mgkg-1), which were administered continuously for 7 d. All groups except the normal control group were given 60 mgkg-1 ANIT at the fifth day. After the last administration, all rats were fasted 24 h and arterial blood were collected to detect the indexes of total bilirubin(TBIL), aspartate aminotransferase(AST), alanine aminotransferase(ALT) and alkaline phosphatase(ALP). The livers were picked for HE staining, immunohistochemistry(IHC) analysis and Western blot(WB), to indentify the protein expressions of CYP3A4 and PXR. RESULTS Compared to model group, low-dose and high-dose matrine decreased TBIL, AST, ALT and ALP significantly; IHC analysis showed that the expression of CYP3A4 in high-dose group was significantly higher than that in model group(P＜0.05). The results of WB showed that the expressions of CYP3A4 and PXR in two matrine groups were significantly higher than that in model group(P＜0.05), however, only CYP3A4 expression in UDCA group was significantly higher than that in model group(P＜0.05). CONCLUSION Matrine could improve cholestatic liver injury in rats and its mechanism might be related to the upregulation of CYP3A4 expression controled by inducing PXR expression.
关键词
苦参碱 /
胆汁淤积 /
CYP3A4 /
孕烷X受体 /
α-萘异硫氰酸酯
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Key words
matrine /
cholestasis /
CYP3A4 /
pregnane X receptor /
α-naphthl isocyanate
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中图分类号：
R965
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参考文献
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脚注
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基金
福建省医学创新课题资助(2016-CX-15)
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