[关键词]
[摘要]
目的 探讨黄连解毒汤有效组分通过激活Caspase-11靶点抗脓毒症的作用机制。方法 实验分为脂多糖(LPS)组、对照组及黄连解毒汤上清液醇洗(HLJDT-7)40、80、120 mg/mL组和黄连解毒汤沉淀水洗(HLJDT-5)40、80、120 mg/mL组。MTT实验检测各组对细胞活力的影响,Western blotting检测Caspase-1和Caspase-11蛋白在黄连解毒汤组分各浓度中的相对表达量。结果 黄连解毒汤组分对LPS处理的RAW264.7细胞进行干预后能够明显的降低LPS对细胞的抑制。当Caspase-11和Caspase-1蛋白敲除后,HLJDT-7 120 mg/mL组和HLJDT-5 40、80、120 mg/mL组对Caspase-11和Caspase-1蛋白表达水平具有明显的抑制作用(P<0.05、0.01)。结论 黄连解毒汤有效组分可降低细胞内的LPS,进而抑制Caspase-11,降低Caspase-11介导的Caspase-1依赖的细胞焦亡作用。
[Key word]
[Abstract]
Objective To investigate the mechanism of effective components of Huanglian Jiedu Decoction against sepsis by activating Caspase-11 target. Methods The experiment was divided into LPS group, control group, HLJDT-7 40, 80, and 120 mg/mL groups, and water extract of Huanglian Jiedu Decoction 40, 80, and 120 mg/mL groups. The effect of each group on cell viability was detected by MTT assay. The relative expression levels of Caspase-1 and Caspase-11 proteins in various concentrations of Huanglian Jiedu Decoction were detected by Western blotting. Results The intervention of Huanglian Jiedu Decoction on LPS-treated RAW264.7 cells could significantly reduce the inhibition rate of LPS on cells. When Caspase-1 and Caspase-11 proteins were knocked out, expressions of Caspase-11 and Caspase-1 proteins in HLJDT-7 120 mg/mL group and HLJDT-5 40, 80, and 120 mg/mL groups were significantly inhibited (P<0.05, 0.01). Conclusion The effective component of Huanglian Jiedu Decoction can reduce LPS in cells and inhibit Caspase-11, and reduce the pyroptosis mediated by Caspase-11 and dependent on Caspase-1.
[中图分类号]
R966
[基金项目]
国家自然科学基金青年科学基金资助项目(81303205);辽宁省高等学校创新人才计划项目(LR2017002);沈阳市中青年科技创新人才支持计划项目(RC170344);沈阳市科技计划项目(18-013-078)