摘要
目的基于microRNA155-SOCS1信号通路探讨“心受气于脾”理论对高脂血症(HLP)脾虚痰浊大鼠血管内皮的保护机制。方法SPF级雄性SD大鼠60只按随机数字表法随机分为正常组,模型组,阿托伐他汀组和健脾祛痰中药低、中、高剂量组。正常组予普通饲料喂饲,模型组、健脾祛痰中药组和阿托伐他汀组均予高脂饲料喂饲联合腹腔注射维生素D3,活动受限,建立高脂血症脾虚痰浊大鼠模型,同时进行药物干预,健脾祛痰中药各组以及阿托伐他汀组分别给予相应的药物灌胃,空白组与模型组给予等量的生理盐水,共计12周。12周后采用紫外分光光度法检测主动脉ATP含量,采用ABTS法检测主动脉总抗氧化能力(T-AOC),采用ELISA法检测各组大鼠血清总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)含量以及氧化低密度脂蛋白(oxidation low lipoprotein,ox-LDL)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-PX)、过氧化氢酶(catalase,CAT)、超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)、细胞间黏附分子1(intercellular cell adhesion molecule 1,ICAM-1)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、大鼠γ干扰素(interferon-γ,IFN-γ)水平。HE染色法检测主动脉脂质沉积;实时荧光定量PCR检测主动脉及斑块内microRNA155、细胞因子信号抑制蛋白1(suppressor of cytokine signaling 1,SOCS1)mRNA的表达。结果与正常组比较,模型组TC、TG、LDL-C、ox-LDL、MDA、ICAM-1、TNF-α、IFN-γ、microRNA155水平显著升高(P<0.01);ATP、T-AOC、GSH-PX、CAT、SOD,SOCS1 mRNA水平显著降低(P<0.01);HE染色主动脉壁明显增厚,内膜不光滑不完整,形成纤维增生性纤维斑块。与模型组比较,健脾祛痰各组大鼠TC、LDL-C、ox-LDL、MDA、ICAM-1、TNF-α、IFN-γ、microRNA155水平降低(P<0.01),ATP、T-AOC、GSH-PX、CAT、SOD、SOCS1 mRNA水平上升(P<0.01),健脾祛痰中、高剂量组TG水平降低(P<0.01)。结论健脾祛痰中药具有良好的保护血管内皮功能的作用,其机制可能是通过激活microRNA155-SOCS1轴信号通路实现的。
Objective To explore the protective mechanism of the theory of“heart receiving Qi from spleen”on the vascular endothelium of hyperlipidemia(HLP)rats with spleen deficiency and phlegm turbid based on the microRNA155-SOCS1 signaling pathway.Methods Sixty SPF male SD rats were divided into normal group,model group,atorvastatin group and low,medium and high dose groups of traditional Chinese medicine by random number table method.The normal group was fed with basic feed.The model group,the strengthening spleen and reducing phlegm groups and atorvastatin group were fed with high-fat feed combined with intraperitoneal injection of vitamin D3,with the activity being restricted,to establish the rat model of hyperlipidemia with spleen deficiency and phlegm turbidity.Simultaneous drug intervention was given.The strengthening spleen and reducing phlegm and the atorvastatin group were given the corresponding drugs by gavage respectively,and the blank group and the model group were given the same amount of normal saline for 12 weeks.After 12 weeks,ATP content in aorta was detected by ultraviolet spectrophotometry.The total antioxidant capacity(T-AOC)of aorta was detected by ABTS method,and serum total cholesterol(TC),triglyceride(TG),low-density lipoprotein(LDL),oxidized low-density lipoprotein(ox-LDL),glutathione peroxidase(GSH-PX),catalase(CAT),superoxide dismutase(SOD),malondialdehyde(MDA),intercellular adhesion molecule 1(ICAM-1),tumor necrosis factorα(TNF-α)and interferonγ(IFN-γ)were measured.Aortic lipid deposition was detected by HE staining.The microRNA155 and SOCS1 mRNA expressions in aorta and plaque were detected by real-time fluorescent quantitative PCR.Results Compared with those of the normal group,the levels of TC,TG,LDL-C,ox-LDL,MDA,ICAM-1,TNF-α,IFN-γand microRNA155 in the model group were significantly increased(P<0.01).The levels of ATP,T-AOC,GSH-PX,CAT,SOD and SOCS1 mRNA were significantly reduced(P<0.01).HE staining showed the aortic wall was obviously thickened and the intima was not smooth and incomplete,forming fibroproliferative fibrous plaques.Compared with those of the model group,the levels of TC,LDL-C,ox-LDL,MDA,ICAM-1,TNF-α,IFN-γand microRNA155 in each strengthening spleen and reducing phlegm group decreased(P<0.01).The levels of ATP,T-AOC,GSH-PX,CAT,SOD and SOCS1 mRNA were significantly increased(P<0.01).The levels of TG in the middle and high dose strengthening spleen and reducing phlegm groups decreased(P<0.01).Conclusion Chinese medicine for strengthening spleen and reducing phlegm has a good protective effect on vascular endothelial function,and its mechanism may be realized by activating the microRNA155-SOCS1 axis signal pathway.
作者
刘悦
赵誉
徐宁阳
韩晓萌
王佳楠
王洋
孔德昭
张艳
卢秉久
LIU Yue;ZHAO Yu;XU Ningyang;HAN Xiaomeng;WANG Jianan;WANG Yang;KONG Dezhao;ZHANG Yan;LU Bingjiu(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Affiliated Hospital of Liaoning University of traditional Chinese Medicine,Shenyang 110032,Liaoning,China)
出处
《中华中医药学刊》
CAS
北大核心
2023年第3期33-37,I0019,共6页
Chinese Archives of Traditional Chinese Medicine
基金
国家重点基础研究发展计划(“973”计划)项目(2013CB531704)
国家自然科学基金项目(81703907)
中国博士后科学基金面上项目(272830)
辽宁省兴辽英才计划项目(XLYC2007057)。
