天麻钩藤饮对自发性高血压大鼠血管衰老中N6-甲基腺苷甲基化的影响  

Effects of Tianma Gouteng Decoction on N6-methyladenosine methylation in vascular aging of spontaneous hypertensive rats

作　　者：姚佳梅[1] 钟博宇 时拥月 秦钦 杨海燕[1] 张丹 王东生[3] 钟广伟[1,3] YAO Jiamei;ZHONG Boyu;SHI Yongyue;QIN Qin;YANG Haiyan;ZHANG Dan;WANG Dongsheng;ZHONG Guangwei(Department of Geriatrics,Xiangya Hospital,Central South University,Changsha 410008,China;Hunan University of Chinese Medicine,Changsha 410208,China;Department of Integrated TCM&Western Medicine,Xiangya Hospital,Central South University,Changsha 410008,China)

机构地区：[1]中南大学湘雅医院老年医学科,长沙410008 [2]湖南中医药大学,长沙410208 [3]中南大学湘雅医院中西医研究所,长沙410008

出　　处：《中华中医药杂志》2025年第3期1098-1105,共8页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81673858);湖南省自然科学基金项目(No.2023JJ30979);湖南省中医药科研计划项目(No.C2022032);湖南省生态环境科研项目(No.HBKYXM-2023017)。

摘　　要：目的:观察天麻钩藤饮对自发性高血压大鼠(SHR)血管衰老中N6-甲基腺苷(m^(6)A)甲基化的影响,探讨其可能的作用机制。方法:10只WKY大鼠为正常组,30只SHR按随机数表法分为模型组、西药对照组和中药组,每组10只,西药对照组和中药组分别给予缬沙坦胶囊和天麻钩藤饮干预8周。观察大鼠血压及形态学改变,β-半乳糖苷酶(SA-β-gal)染色检测血管衰老,比色法检测组织m6A甲基化总体水平;采用实时荧光定量PCR法检测血管组织中甲基转移酶样蛋白3(Mettl3)、甲基转移酶样蛋白14(Mettl14)、肥胖相关蛋白(Fto)、Alkb同系物5(Alkbh5)、YthN6-甲基腺苷RNA结合蛋白1(Ythdf1)、YthN6-甲基腺苷RNA结合蛋白2(Ythdf2)和YTH结构域包含蛋白1(Ythdcl)基因表达及血管衰老相关p21和Sirtl基因表达水平,运用免疫组织化学法检测衰老相关p21和Sirt1蛋白表达,采用免疫组织荧光法检测m°A甲基化相关Mettl3和Ythdc1蛋白表达。结果:与模型组比较,西药对照组和中药组中血压明显下降,内中膜增厚减轻、内中膜厚度/内径降低(P<0.05),SA-β-gal阳性细胞数显著减少(P<0.01),血管组织mA甲基化水平显著升高,药组中p21和Ythdc1表达下降(P<0.01,P<0.05),Sirt1和Mettl3表达增强(P<0.01)。结论:天麻钩藤饮可能通过调控Mettl3与Ythdcl表达,提高m6A甲基化水平,达到延缓血管衰老的作用。Objective:To observe the effects of Tianma Gouteng Decoction on N6 methyladenosine(m^(6)A)methylation during vascular aging in spontaneously hypertensive rats(SHR)and explore its possible mechanism of action.Methods:Ten WKY rats were divided into the normal group,and thirty SHR were randomly divided into model group,western medicine control group and traditional Chinese medicine group,with 10 rats in each group.The western medicine control group and traditional Chinese medicine group were respectively treated with valsartan capsules and Tianma Gouteng Decoction for 8 weeks.The blood pressure was examined and morphology(HE and Masson)and senescence-associatedβ-galactosidase(SA-β-gal)staining were performed to observe the pathological changes of vascular aging.The level of m^(6)A methylation in vascular tissue was detectde by colorimetric method.Real time fluorescence quantitative PCR was used to detect genes expression of methyltransferase like protein 3(Mettl3),methyltransferase like protein 14(Mettl14),obesity related protein(Fto),Alkb homolog 5(Alkbh5),Yth N6 methyladenosine RNA binding protein 1(Ythdf1),Yth N6 methyladenosine RNA binding protein 2(Ythdf2),and YTH domain containing protein 1(Ythdc1),p21 and Sirtl in vascular tissue.Immunohistochemical and immunofifluorescence were used to detect proteins expression of p21,Sirtl,Mettl3 and Ythdc1.Results:After drug intervention,compared with the model group,the western medicine control group and the traditional Chinese medicine group had showed a significant decrease in blood pressure,reduced thickening of the intima-media and thickness/inner diameter of the intima-media(P<0.05),significantly reduced number of SA-β-gal positive cells(P<0.01),and significantly increased m6A methylation level in vascular tissue.The expression of p21 and Ythdc1 were significantly decreased(P<0.01,P<0.05),the expression of Sirtl and Mettl3 were significantly increased(P<0.01).Conclusion:Tianma Gouteng Decoction could delay the vascular aging of SHR,the mechanism may be related to

关 键 词：天麻钩藤饮 自发性高血压大鼠 m^(6)A甲基化 血管衰老 

分 类 号：R285.5[医药卫生—中药学]