半夏白术天麻汤通过AngⅡ/TGF-β1/smad通路干预合并血脂异常自发性高血压病大鼠心肌纤维化  

Banxia Baizhu Tianma Decoction(半夏白术天麻汤)Intervenes in Myocardial Fibrosis in SHR Rats Combined with Dyslipidemia through AngⅡ/TGF-β1/smad Pathway

作　　者：英哲铭 宋囡[1] 曹媛[1] 贾连群[1] 杨关林[1] YING Zheming;SONG Nan;CAO Yuan;JIA Lianqun;YANG Guanin(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847

出　　处：《中华中医药学刊》2024年第9期120-123,I0015-I0019,共9页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82074145);辽宁省自然科学基金项目(2020-MS-228);辽宁省教育厅项目(L202016)。

摘　　要：目的探究半夏白术天麻汤通过调控AngⅡ/TGF-β1/smad通路对高脂喂饲的自发性高血压病大鼠心肌纤维化的影响。方法选取10只Wistar-Kyoto(WKY)大鼠和48只自发性高血压病大鼠(SHR),将SHR大鼠随机分为6组,每组8只,分别为对照组(WKY-ND)、高血压病组(SHR-ND)、高脂高血压病组(SHR-HF)、半夏白术天麻汤低(BDL)、中(BDM)、高(BDH)剂量组和西药(Telmisartan)组。所有大鼠适应性喂养1周后,给予WKY-ND组和SHR-ND组大鼠普通饲料,其余各组给予高脂饲料,喂养8周。造模成功后,给予WKY-ND组、SHR-ND组和SHR-HF组等体积生理盐水,其余各组给予药物干预12周。之后进行大鼠心肌组织取材,采用苏木素-伊红染色(HE)观察大鼠心肌细胞形态、采用马松(Masson)染色观察心肌组织胶原纤维的含量;ELISA法检测血清中AngⅡ、IL-6、TNF-α、NO、ox-LDL、LOX-1、ROS的含量;免疫组织化学方法检测心肌组织α-SMA、ColⅠ、ColⅢ的表达;采用Western blot检测心肌组织TGF-β1、p-smad2、p-smad3、MMP2、TIMP1蛋白表达水平。结果HE、Masson染色显示,给药后心肌细胞大小更均匀,形态更完整,细胞核清晰,胶原纤维面积减少,以半夏白术天麻汤高剂量组给药后改变更为明显;与SHR-ND组相比,SHR-HF组大鼠血清中AngⅡ、IL-6、TNF-α、ox-LDL、LOX-1、ROS水平明显升高,NO水平明显降低;与SHR-HF组相比,各给药组AngⅡ、IL-6、TNF-α、ox-LDL、LOX-1、ROS水平明显降低,NO水平明显升高;与SHR-ND组相比,SHR-HF组大鼠心肌组织中ColⅠ、ColⅢ、α-SMA蛋白水平显著升高;与SHR-HF组相比,各给药组ColⅠ、ColⅢ、α-SMA蛋白水平显著降低;与SHR-ND组相比,SHR-HF组大鼠心肌组织中p-smad2、p-smad3、MMP2的蛋白水平显著升高,TIMP1的蛋白水平显著降低。与SHR-HF组相比,BD组大鼠心肌组织中的TGF-β1、p-smad2、p-smad3、MMP2显著降低,TIMP1显著升高。结论半夏白术天麻汤可通过调节AngⅡ/TGF-β1/smad信号通路,影响高Objective To explore the effect of Banxia Baizhu Tianma Decoction(半夏白术天麻汤,BD)on the myocardial fi-brosis of the spontaneously hypertensive rats(SHR)fed with high fat by regulating angiotensinⅡ(AngⅡ)/transforming growth factor-β1(TGF-β1)/smad pathway.Methods Ten Wistar Kyoto(WKY)rats were being as the control group(WKY-ND)and 48 SHR were were randomly divided into six groups(eight in each group)including hypertension(SHR-ND),high-fat hy-pertension(SHR-HF),low dose(BDL),medium dose(BDM),high dose(BDH)and western medicine(telmisartan)groups.After 1 week of adaptive feeding,all rats in the WKY-ND and SHR-ND groups were given the regular feed,while the other groups were given high-fat feed for 8 weeks.After successful modeling,the WKY-ND,SHR-ND and SHR-HF groups were given the equal volumes of physiological saline,while the other groups were given medication intervention for 12 weeks.And then,the myocardial tissue samples were taken out from the rats.The morphology of myocardial cells was observed by hematoxylin eosin staining(HE),and the contents of collagen fibers in myocardial tissue was observed by Masson staining.ELISA was used to detect the levels of AngⅡ,tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),nitric oxide(NO),oxidative modified low-density lipoprotein(ox-LDL),lectin like oxidized low density lipoprotein receptor 1(LOX-1)and reactive oxygen spe-cies(ROS)in the rat serums.The immunohistochemistry(IHC)method was used to detect the protein expressions of collagen type I(ColⅠ),collagen typeⅢ(ColⅢ)andα-smooth muscle actin(α-SMA).Western blot was used to detect the protein expressions of TGF-β1,phosphorylation signal transduction molecule 2(p-smad2),phosphorylation signal transduction mole-cule 3(p-smad3),matrix metallopeptidase 2(MMP2)and recombinant tissue inhibitors of metalloproteinase 1(TIMP1).Re-sults HE and Masson stainings showed that the myocardial cells were more uniform in size,more complete in shape,clear in nu-cleus,and reduced in collagen fiber area after admi

关 键 词：半夏白术天麻汤 自发性高血压病大鼠 心肌纤维化 血脂异常 TGF-β1/smad通路 

分 类 号：R256.2[医药卫生—中医内科学]