[1]秦润禾,陶 辉,倪世豪,等.microRNA-200b下调DNMT3A的表达对SD大鼠心肌纤维化抑制作用的机制研究[J].中国药理学通报,2018,(10):1465-1470.[doi:10.3969/j.issn.1001-1978.2018.10.027]
 QIN Run-he,TAO Hui,NI Shi-hao,et al.Down-regulation of DNMT3A expression by microRNA-200b inhibits myocardial fibrosis in SD rats[J].Chinese Pharmacological Bulletin,2018,(10):1465-1470.[doi:10.3969/j.issn.1001-1978.2018.10.027]
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microRNA-200b下调DNMT3A的表达对SD大鼠心肌纤维化抑制作用的机制研究()
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《中国药理学通报》[ISSN:/CN:]

卷:
期数:
2018年10期
页码:
1465-1470
栏目:
论著
出版日期:
2018-09-26

文章信息/Info

Title:
Down-regulation of DNMT3A expression by microRNA-200b inhibits myocardial fibrosis in SD rats
文章编号:
1001-1978(2018)10-1465-06
作者:
秦润禾12陶 辉12倪世豪12代 晨12丁季飞12施 鹏12石开虎12
1.安徽医科大学第二附属医院心胸外科,安徽 合肥 230601; 2.安徽医科大学心血管病研究中心,安徽 合肥 230601
Author(s):
QIN Run-he12 TAO Hui12 NI Shi-hao12 DAI Chen12 DING Ji-fei12 SHI Peng12 SHI Kai-hu12
1.Dept of Cardio-Thoracic Surgery, the Second Hospital of Anhui Medical University, Hefei 230601, China; 2.Dept of Cardiovascular Disease Research Center, Anhui Medical University, Hefei 230601, China
关键词:
miR-200b α-SMA Col1A1 DNMT3A 心肌纤维化 心肌成纤维细胞
Keywords:
miR-200b α-SMA Col1A1 DNMT3A myocardial fibrosis cardiac fibroblasts
分类号:
R-332; R322.11; R329.24; R342.22; R342.3; R542.23; R977.6
DOI:
10.3969/j.issn.1001-1978.2018.10.027
文献标志码:
A
摘要:
目的 研究微小RNA200b(miR-200b)靶向下调DNA甲基化转移酶3A(DNMT3A)对SD大鼠心肌纤维化的影响。方法 SD大鼠构建心肌纤维化模型,HE和Masson法观察病理学改变; qRT-PCR检测miR-200b的表达; Western blot法测定α-SMA、Col1A1、DNMT3A的表达。提取SD乳鼠心肌成纤维细胞(CFs),将miR-200b促进剂和抑制剂分别转染于CFs中,同时设立空白及阴性对照组。qRT-PCR检测miR-200b、DNMT3A、Col1A1和α-SMA水平; Western blot测定DNMT3A、Col1A1和α-SMA的蛋白表达; MTT法观察CFs的增殖情况。结果 HE 和Masson染色结果显示,模型组SD大鼠心肌组织胶原纤维明显增多,心肌细胞排列杂乱; qRT-PCR显示miR-200b表达降低; Western blot显示,α-SMA、Col1A1、DNMT3A蛋白水平升高。体外实验qRT-PCR结果显示,α-SMA、Col1A1、DNMT3A 在miR-200b促进剂组中的表达降低,miR-200b抑制剂组则相反; Western blot显示,miR-200b促进剂组中DNMT3A、Col1A1和α-SMA的表达降低,miR-200b抑制剂组结果则相反; MTT结果显示,外源性过表达miR-200b 24、48、72、96 h后,CFs增殖水平降低,而miR-200b抑制剂组结果相反。结论 miR-200b可能通过下调DNMT3A来影响CFs的活化增殖,从而改善心肌纤维化。
Abstract:
Aim To investigate the effect of microRNA200b(miR-200b)targeting down-regulation of DNA methyltransferase 3A(DNMT3A)on myocardial fibrosis in SD rats.Methods In vivo: Myocardial fibrosis model was established in SD rats, HE and Masson method were used to observe the pathological changes, qRT-PCR was used to detect the expression of miR-200b, and Western blot was used to determine the expression of α-SMA, Col1A1 and DNMT3A.In vitro: Cardiomyocyte fibroblasts(CFs)were isolated from SD rats and miR-200b mimics and inhibitors were transfected into CFs.At the same time, blank and negative control groups were established.The levels of miR-200b, DNMT3A, Col1A1 and α-SMA were determined by qRT-PCR.The protein expressions of DNMT3A, Col1A1 and α-SMA were determined by Western blot.The proliferation of CFs was observed by MTT assay.Results In vivo: The results of HE and Masson staining in SD rats in the model group showed that the myocardial tissue showed a significant increase of collagen fibers, and the myocardial cells were arranged disorderly; qRT-PCR showed that the expression of miR-200b decreased; Western blot showed α-SMA, Col1A1, DNMT3A protein The level rises.In vitro: qRT-PCR showed that the expression of α-SMA, Col1A1, and DNMT3A decreased in the miR-200b mimics group, and in the miR-200b inhibitors group.Western blot showed that the expression of DNMT3A, Col1A1, and α-SMA in the miR-200b mimics group was reduced, but the results in the miR-200b inhibitors group were reversed.The results of MTT showed that after exogenous overexpression of miR-200b 24h, 48h, 72h, 96h, the proliferation of CFs decreased, while the results of miR-200b inhibitors group were opposite.Conclusion miR-200b may affect the activation and proliferation of CFs by down-regulating DNMT3A and improve myocardial fibrosis.

参考文献/References:

[1] Nagpal V, Rai R, Place A T, et al.MiR-125b is critical for fibroblast-to-myofibroblast transition and cardiac fibrosis[J].Circulation, 2016,133(3):291-301.
[2] Ng K M, Mok P Y, Butler A W, et al.Amelioration of X-linked related autophagy failure in Danon disease with DNA methylationinhibitor[J].Circulation, 2016, 134(18):1373-89.
[3] Karakikes I, Chaanine A H, Kang S, et al.Therapeutic cardiac-targeted delivery of miR-1 reverses pressure overload-induced cardiac hypertrophy and attenuates pathological remodeling[J].J Am Heart Assoc, 2013, 2: e000078.
[4] Lewis B P, Burge C B, Bartel D P.Conserved seed pairing, often flanked by adenosines,indicates that thousands of human genes are microRNA targets[J].Cell,2005, 120: 15-20.
[5] 汪裕琪,石开虎,吴君旭,等.DNMT3A 和Hyp 在ISO 诱导大鼠心肌纤维化中的表达及相关性研究[J].安徽医科大学学报,2014,49(5):606-9.
[5] Wang Y Q, Shi K H, Wu J X, et, al.Expression and correlation of DNMT3A and Hyp of isoprenaline induced myocardial fibrosis in rats[J].Acta Univ Med Anhui, 2014,49(5):606-9.
[6] 杨健康, 石开虎, 陶 辉,等.DNMT3A与CTGF在AAC致大鼠心肌纤维化中的表达及相关性研究[J].安徽医科大学学报, 2017, 52(9):1302-5.
[6] Yang J K, Shi K H, Tao H, et, al.Expression and correlation of DNMT3A and CTTGF of abdominal aortic coarctation induced myocardial fibrosis in rats[J].Acta Univ Med Anhui, 2017, 52(9):1302-5.
[7] 罗卫民, 罗湘玉, 郭家龙, 等.miR-200b靶向DNMT3A抑制非小细胞肺癌细胞增殖与诱导凋亡[J].天津医药, 2016, 44(8):984-8.
[7] Luo W M, Luo Y X, Guo J L, et al.miR-200b suppresses proliferation and induces apoptosis in non-small cell lung cancer cells by targeting DNMT3A[J].Tianjin Med J, 2016, 44(8):984-8.
[8] 刘小丽, 邢晓为, 黄利华,等.β1肾上腺素受体与心衰、高血压和心肌纤维化之间的关系研究进展[J].中国药理学通报, 2013, 29(12):1640-4.
[8] Liu X L, Xing X W, Huang L H, et al.Research advances on the role of β1-adrenergic receptor in heart failure, hypertension and myocardial fibrosis[J].Chin Pharmacol Bull, 2013, 29(12):1640-4.
[9] 邢晓倩, 徐 健, 吕雄文,等.氯沙坦和辛伐他汀对逆转压力负荷心力衰竭大鼠心室心肌纤维化的作用[J].中国药理学通报, 2009, 25(10):1376-9.
[9] Xing X Q, Xu J, Lyu X W, et al.Combination of simvastatin and losartan attenuates left ventricular myocardial fibrosis after pressure overload state in rats[J].Chin Pharmacol Bull, 2009, 25(10):1376-9.
[10] Baas R, van Teeffelen HAAM, Tjalsma SJD, Timmers H T M.The mixed lineage leukemia 4(MLL4)methyltransferase complex is involved in transforming growth factor beta(TGF-β)-activated gene transcription[J].Transcription, 2018, 9(2):67-74.
[11] Elkouris M, Kontaki H, Stavropoulos A, et al.SET9-mediated regulation of TGF-β signaling links protein methylation to pulmonary fibrosis[J].Cell Rep, 2016,15(12):2733-44.
[12] 彭 丰, 王 敏, 江建新,等.微RNA-200b在胆管癌中的表达及其对癌细胞凋亡与侵袭转移特性的影响[J].中华肝胆外科杂志, 2014, 20(2):123-7.
[12] Peng F, Wang M, Jiang J X, et al.Expression of microRNA-200b in cholangiocarcinoma and its effect on apoptpsis and invasivness of cholangiocarcinoma cells[J].Chin J Hepatobiliary Surg, 2014, 20(2):123-7.
[13] Dou C, Sun L, Jin X, et al.Long non-coding RNA CARLo-5 promotes tumor progression in hepatocellular carcinoma via suppressing miR-200b expression[J].Oncotarget, 2017, 8(41):70172.
[14] Zhou B, Yu J W.A novel identified cirular RNA, circRNA_010567, promotes myocardial fibrosis via suppressing miR-141 by targeting TGF-β1[J].Biochem Biophys Res Commun, 2017,487(4):769-75.

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备注/Memo

备注/Memo:
收稿日期:2018-06-11,修回日期:2018-07-22
基金项目:国家自然科学基金资助项目(No 81570295)
作者简介:秦润禾(1992-),男,硕士生,研究方向:心肌纤维化药理学,E-mail:18855193667@163.com;
石开虎(1963-),男,博士,教授,主任医师,博士生导师,研究方向:心肌纤维化药理学,通讯作者,E-mail: shikaihu@gmail.com
更新日期/Last Update: 2018-08-26