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利用脂多糖(lipopolysaccharide,LPS)介导的药物特异质肝损伤模型,评价壮骨关节丸(Zhuangguguanjie wan,ZGW)诱导的肝损伤并初步探索其机制。采用SD大鼠尾静脉注射LPS(2.8 mg·kg~(-1))方法制备药物特异质肝损伤评价模型,实验分为正常对照组、LPS组、ZGW组和LPS+ZGW组。检测分析血清谷丙转氨酶(alanine aminotransferase,ALT)、天冬氨酸氨基转氨酶(aspartate aminotransferase,AST)、肝脏病理变化(hematoxylin-eosin staining,HE染色)、肝脏组织细胞因子、全血及肝脏组织免疫细胞亚群比例。结果表明,与正常对照组相比,ZGW组和LPS组的ALT、AST、肝脏组织病理学无明显改变(P>0.05),ZGW+LPS组ALT、AST均显著升高(P<0.05),肝脏病理可见肝小叶排列紊乱,肝细胞呈单个或不规则的岛屿状或团块状坏死;肝组织细胞因子分析发现,与正常对照组相比,LPS组和ZGW+LPS组的多种细胞因子发生显著改变(P<0.05或P<0.01),但ZGW+LPS组细胞因子改变的数目和程度更高;血液及肝脏组织免疫细胞亚群分析发现,LPS组血液中CD3~+T cell/lymphocyte比例较对照组显著降低(P<0.01),而肝脏中CD3~+T细胞较对照组显著升高(P<0.05);ZGW+LPS组血液中各免疫细胞较LPS组无明显变化(P>0.05),但肝脏中CD3~+T细胞较LPS组显著升高(P<0.05),表明ZGW联合LPS增加了肝组织中CD3~+T细胞的聚集或活性。上述结果表明机体免疫活化状态下,ZGW进一步促进T淋巴细胞募集至肝脏,导致细胞因子过表达和过度炎症反应,从而诱发药物特异质肝损伤。
The liver injury induced by Zhuangguguanjie wan (ZGW) was evaluated by lipopolysaccharide (LPS) -mediated drug-specific liver injury and its mechanism was explored. The model of drug-specific liver damage was established by tail vein injection of LPS (2.8 mg · kg -1) into SD rats. The experiment was divided into normal control group, LPS group, ZGW group and LPS + ZGW group. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), hematoxylin-eosin staining (HE staining), liver tissue cytokines, whole blood and liver tissue immunity Cell subpopulation ratio. The results showed that compared with the normal control group, ALT, AST and liver histopathology did not change significantly in ZGW group and LPS group (P> 0.05), while ALT and AST in ZGW + LPS group were significantly increased (P <0.05) Liver histopathology showed disorder of hepatic lobules and single or irregular island or mass necrosis of liver cells. Compared with normal control group, liver cytokines showed that LPS group and ZGW + LPS group had more cytokines (P <0.05 or P <0.01). However, the number and the degree of cytokines changed in ZGW + LPS group were higher than those in ZGW + LPS group. The results of immunohistochemistry in blood and liver showed that the levels of CD3 + T cell / lymphocyte (P <0.01), but the level of CD3 ~ + T cells in the liver was significantly higher than that in the control group (P <0.05). There was no significant difference in the number of immune cells in the ZGW + LPS group compared with the LPS group (P> 0.05). However, CD3 + T cells in the liver were significantly increased compared with LPS group (P <0.05), indicating that ZGW combined with LPS increased the aggregation or activity of CD3 + T cells in liver tissue. The above results indicate that ZGW further promotes the recruitment of T lymphocytes to the liver under the condition of immune activation, leading to over-expression of cytokines and over-inflammation, thereby inducing drug-specific liver damage.