文献类型：期刊文献

中文题名：当归红芪超滤物对miR-21-5P靶向调控TGF-β1介导放射性心肌纤维化的干预效应

英文题名：Interventional effect of Radix Angelica Sinensis and Radix Hedysari ultrafiltration on miR-21-5P targeting TGF-β1-mediated radiation-induced myocardial fibrosis

作者：贾成文[1];蒋虎刚[1];高翔[2];韩金晏[1];徐晓东[2];赵信科[2]

第一作者：贾成文

机构：[1]甘肃中医药大学,甘肃兰州730000;[2]甘肃中医药大学附属医院,甘肃兰州730000

第一机构：甘肃中医药大学

年份：2022

卷号：27

期号：11

起止页码：1221

中文期刊名：中国临床药理学与治疗学

外文期刊名：Chinese Journal of Clinical Pharmacology and Therapeutics

收录：CSTPCD;;CSCD:【CSCD_E2021_2022】；

基金：甘肃省青年博士基金(2021QB-080);2021年度教育揭榜挂帅项目(2021jyjbgs-03);2022年陇原青年创新创业人才项目;甘肃省中医药研究中心开放课题计划任务书(zyzx-2020-zx9)。

语种：中文

中文关键词：当归红芪超滤物;放射性心肌纤维化;miR-21-5P;TGF-β1

外文关键词：Radix Angelica Sinensis and Radix Hedysari ultrafiltration;radiation-induced myocardial fibrosis;miR-21-5P;TGF-β1

摘要：目的:揭示miR-21-5P靶向调控TGF-β1是介导心肌成纤维细胞活化的关键机制,明确当归红芪超滤物对miR-21-5P靶向调控TGF-β1机制的干预效应。方法:(1)将细胞随机分为正常组、辐照组,辐照组接受6Gry单次照射,后采用RT-PCR检测miR-21-5P,Western Blot检测α-SMA、TGF-β1的表达;(2)细胞随机分为正常组、辐照组、miR-21-5P _(NC)+辐照组、miR-21-5P _(inhibitor)+辐照组,后采用RT-PCR检测miR-21-5P,Western Blot检测TGF-β1、Smad3、Smad7的表达,免疫荧光染色检测MMP-9、TIMP1表达,EDU染色检测细胞凋亡;(3)将细胞随机分为正常组、辐照组、RAS-RH+辐照组,后采用RT-PCR检测miR-21-5P,Western Blot检测TGF-β1、Smad3、Smad7的表达,免疫荧光染色检测MMP-9、TIMP1表达,EDU染色检测细胞增殖。结果:(1)辐照组miR-21-5P、α-SMA、TGF-β1的表达明显高于正常组(P<0.01),表明X线辐射可诱导心肌成纤维细胞活化,同时miR-21-5P、TGF-β1与心肌纤维化的发生密切相关;(2)miR-21-5P _(inhibitor)+辐照组中miR-21-5P、TGF-β1、Smad3、TIMP1表达明显低于辐照组、miR-21-5P_( NC)+辐照组,MMP-9、Smad7表达明显高于辐照组,EDU染色阳性率降低、细胞数目减少,表明miR-21-5P靶向调控TGF-β1是介导心肌纤维化的关键机制;(3)RAS-RH+辐照组miR-21-5P、α-SMA、TGF-β1、Smad3、TIMP1表达明显低于辐照组,MMP-9、Smad7表达明显高于辐照组,EDU染色阳性率、细胞数目略有降低,表明RAS-RH可通过抑制miR-21-5P靶向调控TGF-β1机制干预放射性心肌纤维化的发生。结论:RAS-RH可通过抑制miR-21-5P靶向调控TGF-β1的机制干预心肌纤维化的发生。
AIM:To reveal that the targeted regulation of TGF-β1 by miR-21-5P is the key mechanism that mediates the activation of myocardial fibroblasts,and to clarify the intervention ofRadix Angelica Sinensis and Radix Hedysari ultrafiltration on the mechanism of miR-21-5P targeting to regulate TGF-β1 effect.METHODS:(1)The cells were randomly divided into normal group and irradiation group.The irradiation group received 6Gry single irradiation,and then RT-PCR was used to detect miR-21-5P,and Western Blot was used to detect the expression ofα-SMA and TGF-β1.(2)The cells were randomly divided into normal group,irradiation group,miR-21-5P_(NC)+irradiation group,miR-21-5P_(inhibitor)+irradiation group,and RT-PCR was used to detect miR-21-5P and Western Blot to detect the expression of TGF-β1,Smad3 and Smad7,the expression of MMP-9 and TIMP1 were detected by immunofluorescence staining,and the apoptosis was detected by EDU staining.(3)The cells were randomly divided into the normal group,the irradiation group,and the RAS-RH+irradiation group.miR-21-5P was detected by RT-PCR,the expressions of TGF-β1,Smad3 and Smad7 were detected by Western Blot,the expressions of MMP-9 and TIMP1 were detected by immunofluorescence staining,and cell proliferation was detected by EDU staining.RESULTS:(1)The expressions of miR-21-5P,α-SMA,and TGF-β1 in the irradiation group were significantly higher than those in the normal group(P<0.05),indicating that X-ray radiation could induce the activation of myocardial fibroblasts,while miR-21-5P and TGF-β1 were closely related to the occurrence of myocardial fibrosis;(2)The expressions of miR-21-5P,TGF-β1,Smad3 and TIMP1 in the miR-21-5P _(inhibitor)+irradiation group were significantly lower than those in the irradiation group,miR-21-5P _(NC)+irradiation group,the expression of MMP-9 and Smad7 was significantly higher than that in the irradiation group,and the positive rate of EDU staining and the number of cells decreased,indicating that miR-21-5P targeted regulation of TGF-β1 was the key to mediating myocardial fibrosis(3)The expressions of miR-21-5P,α-SMA,TGF-β1,Smad3 and TIMP1 in the RAS-RH+irradiation group were significantly lower than those in the irradiation group,and the expressions of MMP-9 and Smad7 were significantly higher than those in the irradiation group.The positive rate of staining and the number of cells decreased slightly,indicating that RAS-RH could interfere with the occurrence of radiation-induced myocardial fibrosis by inhibiting the mechanism of miR-21-5P targeting and regulating TGF-β1.CONCLUSION:RAS-RH can interfere with the occurrence of myocardial fibrosis by inhibiting the mechanism of miR-21-5P targeting and regulating TGF-β1.