红景天总黄酮对大鼠心肌成纤维细胞增殖的抑制作用

doi:10.13481/j.1671-587x.20150618

摘要/Abstract

摘要：

目的:研究红景天总黄酮对转化生长因子β1(TGF-β1)诱导的新生大鼠心肌成纤维细胞(CFB)增殖的抑制作用,探讨红景天总黄酮改善心肌纤维化的作用机制。方法:采用TGF-β1(5 μg·L^-1)诱导大鼠CFB增殖,构建心肌纤维化细胞模型。将正常培养的大鼠CFB分为对照组、模型组和25、50及100 mg·L^-1红景天总黄酮组。给药48 h后,MTT法检测细胞活力,ELISA法检测细胞培养上清液中Ⅰ型胶原蛋白(ColⅠ)和Ⅲ型胶原蛋白(Col Ⅲ)的水平,检测上清液中总超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化物酶(GSH-Px)的活性以及丙二醛(MDA)、谷胱甘肽(GSH)的水平。结果:MTT法检测,与对照组比较,模型组细胞活力明显升高(P<0.01);与模型组,红景天总黄酮各剂量组细胞活力均明显下降(P<0.05)。T-SOD和MDA检测,与对照组比较,模型组细胞T-SOD活性下降(P<0.05),MDA水平明显升高(P<0.05);与模型组比较,50和100 mg·L^-1组细胞T-SOD活性明显升高(P<0.01),25和50 mg·L^-1红景天总黄酮组MDA水平明显下降(P<0.05)。GSH-Px和GSH检测,与对照组比较,模型组细胞GSH-Px活性及GSH水平均明显降低(P<0.01);与模型组比较,红景天总黄酮各剂量组细胞GSH-Px活性和GSH水平均升高(P<0.05)。ColⅠ和Col Ⅲ水平测定,模型组ColⅠ和Col Ⅲ水平明显升高(P<0.01);与模型组比较,红景天总黄酮各剂量组细胞2种蛋白水平均明显下降(P<0.05)。结论:红景天总黄酮可以抑制大鼠CFB的增殖,并可能经抗氧化应激途径改善心肌纤维化。

关键词: 红景天总黄酮, 心肌纤维化, 心肌成纤维细胞, 转化生长因子β1

Abstract:

Objective To investigate the inhibitory effect of Rhodiola total flavonoids on transforming growth factor (TGF)-β1-induced proliferation of cardiac fibroblasts (CFB) in the rats,and to explore its mechanisms of improving myocardial fibrosis. Methods 5 μg·L-1 TGF-β1 was used to induce the proliferation of CFB to build the cell models of myocardial fibrosis.The cultured CFB were divided into control group,model group,and low,medium,high doses(25,50 and 100 mg·L^-1) of Rhodiola total flavonoids groups.The cells were treated for 48 h,the vitalities of cells were detected by MTT, the levels of collagen proteinⅠ (ColⅠ) and collagen proteinⅢ (Col Ⅲ) in supernatant were measured by ELISA,and the activities of total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) were detected, the levels of malondialdehyde (MDA) and glutathione (GSH) were also measured. Results The MTT results indicated that compared with control group,the cell vitality in model group was significantly increased (P<0.01). compared with model group, the cell vitalities of CFB in Rhodiola total flavonoids groups were significantly decreased (P<0.05).Compared with control group,the T-SOD activity in model group was decreased,while the MDA level was significantly increased (P<0.05); compared with model group,the T-SOD activities in 50 and 100 mg·L^-1 Rhodiola total flavonoids groups were increased (P<0.01); the MDA levels in 25 and 50 mg·L^-1 groups were decreased significantly (P<0.05).Compared with control group,the GSH-Px activity and GSH level in model group were significantly decreased (P<0.01). Compared with model group, the GSH-Px activities and GSH levels in Rhodiola total flavonoids groups were increased (P<0.05).Compared with control group, the ColⅠand Col Ⅲ levels in model group were significantly increased (P<0.01); compared with model group, the ColⅠand Col Ⅲ levels in Rhodiola total flavonoids groups were significantly decreased (P<0.05). Conclusion Rhodiola total flavonoids can inhibit the proliferation of rat CFB.The development of myocardial fibrosis may be inhibited by Rhodiola total flavonoids through anti-oxidative stress pathway.

Key words: Rhodiola total flavonoids, myocardial fibrosis, cardiac fibroblasts, transforming growth factor β1

中图分类号:

R285.5

安英, 路倩, 徐丛飞, 王艳春, 韩丽琴. 红景天总黄酮对大鼠心肌成纤维细胞增殖的抑制作用[J]. 吉林大学学报(医学版), 2015, 41(06): 1190-1194.

AN Ying, LU Qian, XU Congfei, WANG Yanchun, HAN Liqin. Inhibitory effect of Rhodiola total flavonoids on proliferation of cardiac fibroblasts in rats[J]. Journal of Jilin University Medicine Edition, 2015, 41(06): 1190-1194.

参考文献

[1] 周凡,陈雪梅,范文玺,等.红景天属植物化学成分研究进展[J].中国中医药信息杂志,2013,20(7):108-109.

[2] 陈雪梅,周凡,马芹,等.不同红景天属植物的药理活性研究进展[J].新疆中医药,2013,31(3):88-90.

[3] 龚晓武,李炳奇,李丹丹,等.红景天黄酮提取及其抗氧化活性研究[J].西北林学院学报,2011,26(3):136-137.

[4] Zhang P,Su J,Mende U.Cross talk between cardiac myocytes and fibroblasts:from multiscale investigative approaches to mechanisms and functional consequences[J].Am J Physiol Heart Circ Physiol,2012,303(12):1385-1396.

[5] Saxena A,Bujak M,Frunza O,et al.CXCR3-independent actions of the CXC chemokine CXCL10 in the infarcted myocardium and in isolated cardiac fibroblasts are mediated through proteoglycans[J].Cardiovasc Res,2014,103(2):217-227.

[6] Nagai T,Anzai T,Kaneko H,et al.C-reactive protein overexpression exacerbates pressure overload-induced cardiac remodeling through enhanced inflammatory response[J].Hypertension,2011,57(2):208-215.

[7] Chen TL,Liao JW,Chan WH,et al.Induction of cardiac fibrosis and transforming growth factor-β1 by motorcycle exhaust in rats[J].Inhal Toxicol,2013,25(9):525-535.

[8] Gong W,Yan M,Chen J,et al.Chronic inhibition of cyclic guanosine monophosphate-specific phosphodiesterase 5 prevented cardiac fibrosis through inhibition of transforming growth factor β-induced Smad signaling[J].Front Med,2014,8(4):445-455.

[9] 李永胜,严丽,雍永权,等.丹参酮ⅡA对高血压大鼠肥厚心肌TGF-β1/Smads信号通路的影响[J].中国中西医结合杂志,2010,30(5):499-503.

[10] Panossian A,Wikman G,Sarris J.Rosenroot(Rhodiola rosea):traditional use,chemical composition,pharmacology and clinical efficacy[J].Phytonmedicine,2010,17(7):481-493.

[11] 刘敏,陈志武.槲皮素-3-葡萄糖苷对心肌缺血缺氧损伤的保护作用[J].安徽医科大学学报,2008,46(3):683-685.

[12] 李贵芝,周红,房彩霞,等.法舒地尔抑制氧化应激反应减轻2型糖尿病大鼠心肌纤维化[J].基础医学与临床,2014,34(2):216-221.

[13] van Deel ED,Lu Z,Xu X,et al.Extracellular superoxide dismutase protects the heart against oxidative stress and hypertrophy after myocardial infarction[J].Free Radic Biol Med,2008,44(7):1305-1313.

[14] Kliment CR,Suliman HB,Tobolewski JM,et al.Extracellular superoxide dismutase regulates cardiac function and fibrosis[J].J Mol Cell Cardiol,2009,47(5):730-742.

[15] 郭蕴苹,杨硕,殷睿,等.滇橄榄的总黄酮含量和SOD活性研究[J].昆明医科大学学报,2012,33(6):35-38.

[16] Jia N,Dong P,Ye Y,et al.Allopurinol attenuates oxidative stress and cardiac fibrosis in angiotensin Ⅱ-induced cardiac diastolic dysfunction[J].Cardiovasc Ther,2012,30(2):117-123.

[17] 韩亚男,程新宇,侯俊玲,等.大孔树脂纯化甘草地上部分总黄酮的工艺[J].吉林中医药,2014,34(1):82-86.