摘要
目的探究微小RNA(miR)-363调控1-磷酸鞘氨醇受体1(S1PR1)/信号传导及转录激活蛋白3(STAT3)对风湿性心脏病(RHD)的影响及作用。方法选择雌性Lewis大鼠60只,随机分为对照组(Con组)、Con组1、RHD组、RHD组1、阴性对照组(转染antagomir-NC)和拮抗剂组(转染miR-363 antagomir),每组10只。后4组通过皮下注射灭活A组乙型溶血性链球菌诱导建立RHD模型,阴性对照组和拮抗剂组分别于尾静脉注射体内导入10 mg/kg的antagomir-NC和miR-363 antagomir,每周2次,持续7周。取心脏瓣膜组织,苏木精-伊红染色检测组织病理改变,用Ⅲ型胶原与Ⅰ型胶原比值和Ⅲ型胶原α1、成纤维细胞特异蛋白1(FSP1)评估心脏瓣膜纤维化,ELISA检测血清白细胞介素(IL)-6、IL-17、IL-21水平,RT-PCR检测miR-363和S1PR1基因表达。用荧光素酶实验检测miR-363与S1PR1靶向作用关系,Western blot检测S1PR1和STAT3蛋白表达。结果与Con组比较,RHD组大鼠组织出现明显的病理性改变和纤维化,Ⅱ型胶原与Ⅰ型胶原比值、Ⅲ型胶原α1和FSP1表达水平升高,miR-363表达水平升高(2.24±0.33 vs 1.00±0.20),S1PR1表达水平下降(0.24±0.09 vs 0.86±0.11,P<0.01)。miR-363可靶向作用于S1PR1。与RHD组1比较,阴性对照组各项无明显变化(P>0.05),拮抗剂组肌酸激酶同工酶和血清肌钙蛋白I水平降低,组织病理改变明显减轻,Ⅱ型胶原与Ⅰ型胶原比值、Ⅲ型胶原α1和FSP1表达降低(12.10±2.92 vs 18.05±3.52、3.13±0.30 vs 4.20±0.41和1.72±0.17 vs 2.37±0.19),IL-6、IL-17和IL-21水平降低,S1PR1基因和蛋白表达水平升高,磷酸化STAT3/STAT3比值降低(P<0.05)。结论miR-363可靶向作用于S1PR1,调控S1PR1/STAT3通路的激活,减轻RHD引起的心脏瓣膜损伤。
Objective To study the effect of miR-363 on rheumatic heart disease(RHD)by regulating the S1 PR1/STAT3.Methods Sixty female Lewis rats were divided into control group and control group 1,RHD group and RHD group 1,negative control group and antagonist group(10 in each group).A RHD model of rats was established by subcutaneous injection with inactivated group B hemolytic streptococcus for RHD group,RHD group 1,negative control group and antagonist group.The animals in negative control group and antagonist group were killed and cardiac valve tisse samples were taken after they received intravenous injection with antagomir NC or miR-363 antagomir for 7 weeks.The pathological changes of cardiac valve tissue were detected with HE staining.The fibrosis of cardiac valve tissue was assessed according to the COLⅢ/Ⅰratio,COLⅢα1 and FSP1.The serum levels of IL-6,IL-17 and IL-21 were measured by ELISA.The expressions of miR-363 and S1 PR1 genes were detected by RT-PCR.Results The pathological changes and fibrosis of cardiac valve tissue were more significant,the COLⅢ/Ⅰratio and expression levels of COLⅢα1 and FSP1 were significantly higher in RHD group than in control group(P<0.01).The expression level of miR-363 was significantly higher while that of S1 PR1 was significantly lower in RHD group than in control group(P<0.01).The miR-363 can target the S1 PR1.The heart rate was significantly higher,the serum CK-MB and cTnI levels were significantly lower,the pathological changes of cardiac valve tissue were significantly milder,the COLⅢ/Ⅰratio and expression levels of COLⅢα1 and FSP1 were significantly lower(12.10±2.92 vs 18.05±3.52,3.13±0.30 vs 4.20±0.41,1.72±0.17 vs 2.37±0.19),the expression levels of IL-6,IL-17 and IL-21 were significantly lower and those of S1 PR1 gene and protein were significantly higher,the phosphorylation rate of STAT3 was significantly lower in antagonist group than in other groups(P<0.05).Conclusion miR-363 can target the S1 PR1,regulate the activation of S1 PR1/STAT3 pathways and reduce RHD-induced heart valve damage in rats.
作者
杨兵
陈博
陈健
李龙山
冯建钦
李娜
Yang Bing;Chen Bo;Chen Jian;Li Longshan;Feng Jianqin;Li Na(Department of Cardiology,Jinan No.3 People's Hospital,Jinan 250132,Shandong Province,China)
出处
《中华老年心脑血管病杂志》
北大核心
2021年第11期1200-1204,共5页
Chinese Journal of Geriatric Heart,Brain and Vessel Diseases
基金
山东省医药卫生科技发展计划项目(2017WS433)。
