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柴胡皂苷b_2对脂多糖/氨基半乳糖诱导的急性肝损伤小鼠炎症及能量代谢的影响

[Effects of saikosaponin b_2 on inflammation and energy metabolism in mice with acute liver injury induced by LPS/GalN].

作者信息

You Man, Li Rui-Fang, Gao Zi-Han, Li Yuan-Ye, Liu Wei-Yi, Wang Jian-Gang, Wang Hong-Wei, Li San-Qiang

机构信息

Medical College,Henan University of Science and Technology Luoyang 471023,China.

School of Nursing,Henan University of Science and Technology Luoyang 471023,China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2019 Jul;44(14):2966-2971. doi: 10.19540/j.cnki.cjcmm.20190521.415.

Abstract

To study the effects of saikosaponin b2( SS-b2) on inflammatory factors and energy metabolism against lipopolysaccharide/galactosamine( LPS/Gal N) induced acute liver injury in mice. Mice were randomly divided into normal group( equal amount of normal saline),model group( 100 g·kg(-1) LPS and 400 mg·kg(-1) Gal N),low,medium,high dose group of SS-b2( SS-b25,10,20 mg·kg(-1)·d-1) and positive control group( dexamethasone,10 mg·kg(-1)). All of the groups except for the normal group were treated with LPS/Gal N though intraperitoneally injection to establish the acute liver injury model. The organ indexes were calculated. The levels of serum transaminases( ALT and AST) and the activities of ATPase( Na+-K+-ATPase,Ca2+-Mg2+-ATPase) in liver were detected. The activity of tumor necrosis factor-α( TNF-α),interleukin-1β( IL-1β) and interleukin-6( IL-6) were determined by the enzyme-linked immunosorbent assay( ELISA). The contents of lactate dehydrogenase( LDH) in liver were determined by micro-enzyme method. HE staining was used to observe the histopathological changes of the liver. Histochemical method was used to investigate the protein expression of liver lactate dehydrogenase-A( LDH-A). The protein expressions of Sirt-6 and NF-κB in the liver were detected by Western blot. According to the results,compared with the model group,there were significant changes in organ indexes in the high-dose group of SS-b2( P<0. 05). The level of ALT,AST,TNF-α,IL-1β,IL-6 and the activities of LDH in serum of mice with liver injury were significantly reduced in the medium and high dose groups of SS-b2( P<0. 01). With the increase of the concentration of SS-b2,the range of hepatic lesions and the damage in mice decreased. The activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase in liver of mice were significantly enhanced in each dose group( P<0. 01). The expression of NF-κB in liver tissues was significantly down-regulated in the medium and high dose group( P<0. 01). Meanwhile,the expression of Sirt-6 protein in the liver of mice with acute liver injury was significantly increased in each dose group( P<0. 01).In summary,SS-b2 has a significant protective effect on LPS/Gal N-induced acute liver injury in mice,which may be related to the down-regulation of NF-κB protein expression and up-regulation of Sirt-6 protein expression to improve inflammatory injury and energy metabolism.

摘要

研究柴胡皂苷b2(SS-b2)对脂多糖/氨基半乳糖(LPS/Gal N)诱导的小鼠急性肝损伤炎症因子及能量代谢的影响。将小鼠随机分为正常组(等量生理盐水)、模型组(100 g·kg⁻¹ LPS和400 mg·kg⁻¹ Gal N)、SS-b2低、中、高剂量组(SS-b2 5、10、20 mg·kg⁻¹·d⁻¹)和阳性对照组(地塞米松,10 mg·kg⁻¹)。除正常组外,其余各组均通过腹腔注射LPS/Gal N建立急性肝损伤模型。计算脏器指数。检测血清转氨酶(ALT和AST)水平及肝脏中ATP酶(Na⁺-K⁺-ATP酶、Ca²⁺-Mg²⁺-ATP酶)活性。采用酶联免疫吸附测定法(ELISA)测定肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)活性。采用微量酶法测定肝脏中乳酸脱氢酶(LDH)含量。采用HE染色观察肝脏组织病理学变化。采用组织化学方法检测肝脏乳酸脱氢酶-A(LDH-A)蛋白表达。采用蛋白质免疫印迹法检测肝脏中Sirt-6和NF-κB蛋白表达。结果显示,与模型组相比,SS-b2高剂量组脏器指数有显著变化(P<0.05)。SS-b2中、高剂量组肝损伤小鼠血清中ALT、AST、TNF-α、IL-1β、IL-6水平及LDH活性显著降低(P<0.01)。随着SS-b2浓度升高,小鼠肝脏病变范围及损伤程度减小。各剂量组小鼠肝脏中Na⁺-K⁺-ATP酶和Ca²⁺-Mg²⁺-ATP酶活性显著增强(P<0.01)。中、高剂量组肝脏组织中NF-κB表达显著下调(P<0.01)。同时,各剂量组急性肝损伤小鼠肝脏中Sirt-6蛋白表达显著增加(P<0.01)。综上所述,SS-b2对LPS/Gal N诱导的小鼠急性肝损伤具有显著保护作用,其机制可能与下调NF-κB蛋白表达、上调Sirt-6蛋白表达从而改善炎症损伤及能量代谢有关。

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