补阳还五汤苷类组分调控动脉粥样硬化炎性反应及脂质代谢的作用机制  

Mechanism of glycosides of Buyang Huanwu Decoction on regulating atherosclerotic inflammation and lipid metabolism

作　　者：傅馨莹 杨仁义[1] 孙正骥 李艳玲[1] 龙清吟 李俊熙 吴露[2] 张伟[1] FU Xin-ying;YANG Ren-yi;SUN Zheng-ji;LI Yan-ling;LONG Qing-yin;LI Jun-xi;WU Lu;ZHANG Wei(Hunan University of Traditional Chinese Medicine,Changsha 410208,China;Liuyang Hospital of Traditional Chinese Medicine,Liuyang 410300,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]浏阳市中医医院,湖南浏阳410300

出　　处：《中草药》2021年第14期4221-4231,共11页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81774032);湖南省自然科学杰出青年基金资助项目(2020JJ2024);湖南省自然科学青年基金资助项目(2018JJ3391);湖南省教育厅科学研究重点项目(19A374);湖南中医药大学研究生培养质量工程专项(2020CX62)。

摘　　要：目的采用网络药理学结合体外实验验证的方法,探讨补阳还五汤苷类组分治疗动脉粥样硬化(atherosclerosis,AS)的作用机制。方法采用网络药理学方法对补阳还五汤苷类组分与阿托伐他汀治疗AS的靶点和通路进行预测;SD大鼠随机分为对照组、补阳还五汤(10.8 g/kg)组、苷类组分(0.64 g/kg)组和阿托伐他汀(0.9 mg/kg)组,各给药组ig相应药物,2次/d,连续7次,制备空白血清、补阳还五汤血清、苷类组分血清及阿托伐他汀血清。采用试剂盒考察含药血清对氧化型低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)诱导的小鼠单核巨噬细胞RAW264.7脂质含量的影响;采用油红O染色考察含药血清对ox-LDL诱导的RAW264.7细胞泡沫细胞形成率的影响;采用ELISA试剂盒观察含药血清对ox-LDL诱导的RAW264.7细胞上清液肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白细胞介素-6(interleukin-6,IL-6)水平的影响;采用Western blotting考察含药血清对ox-LDL诱导的RAW264.7细胞Janus蛋白酪氨酸激酶2(Janus kinase-2,JAK2)、磷酸化JAK2(p-JAK2)、信号转导和转录激活因子3(signaltransducer and activator of transcription 3,STAT3)和p-STAT3蛋白表达的影响;采用qRT-PCR考察含药血清对ox-LDL诱导的RAW264.7细胞JAK2和STAT3 mRNA表达的影响。结果与模型组比较,各给药组细胞内脂质沉积减少,RAW264.7细胞吞噬脂质受到抑制,泡沫化细胞数量减少,上清液中TNF-α和IL-6水平显著降低(P<0.01),细胞内总胆固醇(total cholesterol,TC)、游离胆固醇(free cholesterol,FC)和胆固醇酯(cholesteryl ester,CE)含量显著减少(P<0.01),JAK2/STAT3通路相关蛋白及m RNA表达水平显著降低(P<0.01)。结论补阳还五汤苷类组分是治疗AS的主要药效物质,其作用机制可能与降低细胞内脂质沉积、抑制JAK2/STAT3信号通路、抑制炎性因子分泌有关。Objective To explore the mechanism of Buyang Huanwu Decoction(补阳还五汤) glycosides on treatment of atherosclerosis(AS) by network pharmacology combined with in vitro experiment. Methods Network pharmacology method was used to predict the target and pathway of Buyang Huanwu Decoction glycosides and atorvastatin in the treatment of AS. SD rats were randomly divided into control group, Buyang Huanwu Decoction(10.8 g/kg) group, glycoside component(0.64 g/kg) group and atorvastatin(0.9 mg/kg) group, rats in each administration group were ig corresponding drugs, twice one day for 7 d. The blank serum, Buyang Huanwu Decoction serum, glycoside component serum and atorvastatin serum were prepared. Kit was used to investigate the effect of drug-containing serum on lipid content of RAW264.7 cells induced by oxidized low-density lipoprotein(ox-LDL);Oil red O staining was used to investigate the effect of drug-containing serum on foam cell formation rate of RAW264.7 cells induced by ox-LDL;ELISA kit was used to investigate the effect of drug-containing serum on levels of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) in the supernatant of RAW264.7 cells induced by ox-LDL;Western blotting was used to investigate the effect of drug-containing serum on expressions of Janus protein tyrosine kinase 2(JAK2), phosphorylated JAK2(p-JAK2), signal transducer and activator of transcription 3(STAT3) and p-STAT3 in RAW264.7 cells induced by ox-LDL;qRT-PCR was used to investigate the effect of drug-containing serum on JAK2 and STAT3 m RNA expressions in RAW264.7 cells induced by ox-LDL. Results Compared with model group, the intracellular lipid deposition of each administration group was reduced, the phagocytic lipid of RAW264.7 cells was inhibited, the number of foamed cells was reduced;Levels of TNF-α and IL-6 in the supernatant were significantly reduced(P < 0.01);Intracellular total cholesterol(TC), free cholesterol(FC) and cholesterol ester(CE) content were significantly reduced(P < 0.01);Expressions of JAK2/STAT3 pa

关 键 词：补阳还五汤 苷类组分 阿托伐他汀 动脉粥样硬化 炎性反应 脂质代谢 网络药理学 

分 类 号：R285.5[医药卫生—中药学]