清脂通脉颗粒对动脉粥样硬化病变NF-κB信号通路的影响

Effect of Qingzhi Tongmai Granules(清脂通脉颗粒)on NF-κB Signaling Pathway in Atherosclerotic Lesions

目的考察清脂通脉颗粒药物对细胞活性的影响,探究其抗动脉粥样硬化的作用及机制。方法通过CCK-8法实验判断清脂通脉颗粒的细胞毒性;通过PMA诱导的人THP-1细胞实验,ox-LDL(氧化低密度脂蛋白)诱导后研究清脂通脉颗粒对脂质吞噬的影响;进行Westblot实验进行清脂通脉颗粒对蛋白影响的测定;通过NF-KB通路抑制剂JSH-23联合给予验证清脂通脉颗粒作用靶点。结果CCK-8法实验研究结果表明:不同浓度的清脂通脉颗粒对THP-1细胞活性均无影响;模型组细胞总胆固醇显著增加(P<0.001);清脂通脉颗粒不同浓度组细胞中总胆固醇含量有减少的趋势,且100、200μg/mL组内胆固醇含量更低,两组间无显著差异;清脂通脉颗粒可减少NF-κB信号通路中半胱氨酸蛋白酶3(caspase 3,CASP3)、半胱氨酸蛋白酶9(caspase 9,CASP-9)、核转录因子κB-p65蛋白(p65)含量表达(P<0.01),减少B细胞淋巴相关X蛋白(Bax)蛋白含量表达(P<0.05),增加B细胞淋巴瘤-2(Bcl-2)蛋白含量表达(P<0.01);清脂通脉颗粒作用于巨噬细胞,与NF-κB通路抑制剂JSH-23作用靶点相同。结论细胞水平验证清脂通脉颗粒对其活性无影响、无毒性、安全可靠;清脂通脉颗粒可吞噬脂质、减少巨噬细胞中胆固醇含量;清脂通脉颗粒可通过调控NF-κB通路内相关分子蛋白表达发挥抗AS(动脉粥样硬化)作用;清脂通脉颗粒与NF-κB通路抑制剂JSH-23作用相同。

Objective To investigate the effect of Qingzhi Tongmai Granules(清脂通脉颗粒)on cell activity and explore its anti-atherosclerosis effect and mechanism.Methods The cytotoxicity of Qingzhi Tongmai Granules was determined by CCK-8 assay.The effect of Qingzhi Tongmai Granules on lipid phagocytosis was studied by human THP-1 cells induced by PMA and ox-LDL.Western Blot test was carried out to determine the effect of Qingzhi Tongmai Granules on protein.The target of Qing-zhi Tongmai Granules was verified by the combination of NF-κB pathway inhibitor JSH-23.Results CCK-8 method showed that Qingzhi Tongmai Granules of different concentrations had no effect on THP-1 cell activity.The total cholesterol in the mod-el group was significantly increased(P<0.001).The total cholesterol content in cells of different concentrations of Qingzhi Tongmai Granules showed a decreasing trend,and the cholesterol content in 100μg/mL and 200μg/mL groups was lower,and there was no significant difference between the two groups.Qingzhi Tongmai Granules could reduce the expressions of caspase 3(CASP-3),caspase 9(CASP-9),P65 and protein in NF-κB signaling pathway(P<0.01)and the expression of Bax protein(P<0.05),and increase the expression of Bcl-2 protein(P<0.01).Qingzhi Tongmai Granules acted on macrophages and tar-get the same as NF-κB pathway inhibitor JSH-23.Conclusions At the cell level,Qingzhi Tongmai Granules have no effect on its activity,with safety and reliability but no toxicity.Qingzhi Tongmai Granules have the effect of phagocytic lipid and can reduce the cholesterol content in macrophages.Qingzhi Tongmai Granules can play an anti-atherosclerosis role by regulating the ex-pressions of related molecular proteins in NF-κB pathway.Qingzhi Tongmai Granules have the same effect as NF-κB pathway inhibitor JSH-23.