摘要
目的基于微小RNA-126(miR-126)/解整合素金属蛋白酶9(ADAM9)信号通路研究五脏温阳化瘀汤抗大鼠动脉粥样硬化的作用。方法取SPF级SD雄性大鼠38只,随机选择6只大鼠设为空白组,剩余32只大鼠通过腹腔注射维生素D 3注射液联合高脂饲料喂养方法制备动脉粥样硬化模型。造模6周后将造模成功的大鼠随机分为模型组,阿托伐他汀组及五脏温阳化瘀方低、中、高剂量组,每组6只。空白组和模型组给予生理盐水灌胃,阿托伐他汀组给予阿托伐他汀钙片27.3 mg/(kg·d)灌胃,五脏温阳化瘀方低、中、高剂量组分别给予五脏温阳化瘀汤1.25 g/(kg·d)、2.5 g/(kg·d)、5 g/(kg·d)灌胃,均1次/d,连续灌胃12周。灌胃结束后,采用ELISA法检测血清总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)水平,HE染色观察主动脉病理形态,qRT-PCR法检测主动脉组织中miR126及ADAM9 mRNA表达情况,Western blot法检测主动脉组织中ADAM9蛋白表达情况。结果与空白组比较,模型组大鼠血清TC、TG、LDL-C、TNF-α、IL-1β水平和主动脉组织中ADAM9 mRNA及蛋白相对表达量均明显升高(P均<0.05),血清HDL-C水平和主动脉组织中miR-126相对表达量均明显降低(P均<0.05);与模型组比较,各药物组大鼠血清TC、TG、LDL-C、TNF-α、IL-1β水平和主动脉组织中ADAM9 mRNA及蛋白相对表达量均明显降低(P均<0.05),血清HDL-C水平和主动脉组织中miR-126相对表达量均明显升高(P均<0.05);与五脏温阳化瘀方低、中剂量组比较,阿托伐他汀组与五脏温阳化瘀方高剂量组大鼠血清TC、TG、LDL-C、TNF-α、IL-1β水平和主动脉组织中ADAM9 mRNA及蛋白相对表达量均更低(P均<0.05),血清HDL-C水平和主动脉组织中miR-126相对表达量均更高(P均<0.05),五脏温阳化瘀方高剂量组与阿托伐他汀组各指标比较差异均无统计学意义(P均>0.05)。模型组大鼠主动脉血管内皮缺损、内膜增厚,大量泡沫细胞聚集和炎细胞浸润;各药物组大鼠主动脉病理改变较模型组轻,泡沫细胞及炎性细胞浸润少。结论五脏温阳化瘀方可以通过激活miR-126/ADAM9信号通路,降低炎症因子水平,减轻动脉粥样硬化大鼠炎症反应,从而发挥抗动脉粥样硬化作用。
Objective It is to study the anti atherosclerosis effect and mechanism of decoction for warming yang and removing stasis in five organs in rats based on miR-126/A distegrinin and ametalloprotease 9(ADAM9)signal pathway.Methods Thirty-eight SPF-grade SD male rats were selected,in which 6 rats were randomly selected as the blank group,and the remaining 32 rats were given vitamin D3 injection by intraperitoneal injection and fed with high-fat chow to establish models of atherosclerosis.After modeling for 6 wees,the successfully modeled rats were randomly divided into model group,atorvastatin group,and low,medium and high dose groups of decoction for warming yang and removing stasis in five organs,with 6 rats in each group.The blank group and model control group were given physiological saline by gavage,the atorvastatin group was given atorvastatin calcium tablets 27.3 mg/(kg·d)by gavage,and the low,medium,and high dose groups of decoction for warming yang and removing stasis in five organs were respectively given decoction for warming yang and removing stasis in five organs 1.25,2.5,and 5 g/(kg·d)by gavage,all once daily,continuously treated for 12 weeks.At the end of gavage,the serum levels of total cholesterol(TC),triacylglycerol(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),tumor necrosis factor-α(TNF-α),and interleukin-1β(IL-1β)were determined by ELISA,the pathological morphology of aorta was observed by HE staining,and the expressions of miR126 and ADAM9 mRNA in aortic tissues were detected by qRT-PCR,and the expression of ADAM9 protein in aortic tissues was detected by Western blot.Results Compared with the blank group,the serum levels of TC,TG,LDL-C,TNF-α,IL-1βand relative expressions of ADAM9 mRNA and protein in aortic tissues of rats in the model group were significantly increased(all P<0.05),and the serum level of HDL-C and relative expression of miR-126 in aortic tissues were significantly decreased(all P<0.05);compared with the model group,the serum levels of TC,TG,LDL-C,TNF-α,IL-1βand relative expressions of ADAM9 mRNA and protein in aortic tissues of rats in the treatment groups were significantly decreased(all P<0.05),and the serum level of HDL-C and relative expression of miR-126 in aortic tissues were significantly increased(all P<0.05);compared with the low and medium dose groups of decoction for warming yang and removing stasis in five organs,the serum levels of TC,TG,LDL-C,TNF-α,IL-1βand relative expressions of ADAM9 mRNA and protein in aortic tissues of rats in the atorvastatin group and high dose group of decoction for warming yang and removing stasis in five organs were more lower(all P<0.05),and the serum level of HDL-C and relative expression of miR-126 in aortic tissues were more higher(all P<0.05),the differences in all indicators between the atorvastatin group and high dose group of decoction for warming yang and removing stasis in five organs were not statistically significant(all P>0.05).The rats in the model group had endothelial defects in the aortic vessels,thickening of the intima,and a large number of foam cell aggregates and inflammatory cell infiltration;the pathological changes in the aorta of rats in each treatment group were milder than those in the model group,and there were fewer foam cells and inflammatory cell infiltration.Conclusion Decoction for warming yang and removing stasis in five organs can can exert anti-atherosclerotic effect by activating miR-126/ADAM9 signaling pathway,decreasing the level of inflammatory factors,and reducing the inflammatory response in atherosclerotic rats.
作者
姜明贺
朱欢欢
张鼎
李方存
宋晨曦
陈炜
胡跃强
JIANG Minghe;ZHU Huanhuan;ZHANG Ding;LI Fangcun;SONG Chenxi;CHEN Wei;HU Yueqiang(Guangxi University of Chinese Medicine,Nanning 530001,Guangxi,China;Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530011,Guangxi,China;The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,Guangxi,China)
出处
《现代中西医结合杂志》
CAS
2023年第24期3364-3370,共7页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
国家自然科学基金地区基金项目(82260904)
桂派杏林拔尖人才项目(04B2300807)
广西中医药大学2022年研究生创新教育计划项目(YCBXJ2022012)。
