基于ROS-MAPK信号通路探讨桃叶珊瑚苷抗心肌纤维化作用
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摘要
目的研究桃叶珊瑚苷(AU)对血管紧张素Ⅱ(AngⅡ)诱导的大鼠心肌成纤维细胞(CFs)增殖的影响及作用机制。方法采用胰酶消化法及差速贴壁法培养大鼠CFs,实验分为空白对照组、AngⅡ(1×10~(-6) mol/L)干预组、低浓度AU干预组[AngⅡ(1×10~(-6) mol/L)+AU(1μmol/L)]、中浓度AU干预组[AngⅡ(1×10~(-6) mol/L)+AU(10μmol/L)]、高浓度AU干预组[AngⅡ(1×10~(-6) mol/L)+AU(100μmol/L)]。用CCK8法测定细胞增殖活性,酶联免疫吸附法(ELISA)测定细胞上清中Ⅰ、Ⅲ型胶原含量,荧光酶标仪检测活性氧(ROS)水平,Western blot法检测p38MAPK和p-p38MAPK蛋白表达。结果与AngⅡ干预组比较,中、高浓度AU干预组CFs增殖和p38MAPK磷酸化被明显抑制(P <0.05)。与AngⅡ干预组比较,低、中、高浓度AU干预组ROS、Ⅰ、Ⅲ型胶原的生成明显减少(P <0.05)。结论 AU可通过ROS-MAPK通路途径抑制AngⅡ诱导的CFs增殖和胶原生成。
Objective To investigate the effect and mechanism of aucubin(AU) on cardiac fibroblasts(CFs) proliferation induced by angiotension Ⅱ(Ang Ⅱ). Methods Rat CFs were cultured by the methods of trypsin digestion and differential adhesion. They were divided into blank control group, Ang Ⅱ(1×10~(-6) mol/L) intervention group, low concentration of AU intervention group [Ang Ⅱ(1×10~(-6) mol/L) + AU(1 μmol/L)], medium concentration of AU intervention group [AngⅡ(1×10~(-6) mol/L) + AU(10 μmol/L)], high concentration of AU intervention group [Ang Ⅱ(1×10~(-6) mol/L) + AU(100 μmol/L)].CCK8 method was used to test cell proliferation activity, collagen type Ⅰ, Ⅲ content in cell supernatant were determed by enzyme linked immunosorbent(ELISA), fluorescence enzyme standard instrument was used to detect reactive oxygen species(ROS) level, Western blot method was used to detect p38 MAPK and p-p38 MAPK protein expression.Results Compared with Ang Ⅱ intervention group, CFs proliferation and p38 MAPK phosphorylation were significantly inhibited in the medium and high concentration AU intervention groups(P < 0.05). Compared with Ang Ⅱ intervention group, low, medium and high concentration of AU intervention group ROS, the production of collagen type Ⅰ, Ⅲ decreased significantly(P < 0.05). Conclusion AU can restrain by ROS-MAPK pathway Ang Ⅱ induction of CFs proliferation and collagen formation.
Objective To investigate the effect and mechanism of aucubin(AU) on cardiac fibroblasts(CFs) proliferation induced by angiotension Ⅱ(Ang Ⅱ). Methods Rat CFs were cultured by the methods of trypsin digestion and differential adhesion. They were divided into blank control group, Ang Ⅱ(1×10~(-6) mol/L) intervention group, low concentration of AU intervention group [Ang Ⅱ(1×10~(-6) mol/L) + AU(1 μmol/L)], medium concentration of AU intervention group [AngⅡ(1×10~(-6) mol/L) + AU(10 μmol/L)], high concentration of AU intervention group [Ang Ⅱ(1×10~(-6) mol/L) + AU(100 μmol/L)].CCK8 method was used to test cell proliferation activity, collagen type Ⅰ, Ⅲ content in cell supernatant were determed by enzyme linked immunosorbent(ELISA), fluorescence enzyme standard instrument was used to detect reactive oxygen species(ROS) level, Western blot method was used to detect p38 MAPK and p-p38 MAPK protein expression.Results Compared with Ang Ⅱ intervention group, CFs proliferation and p38 MAPK phosphorylation were significantly inhibited in the medium and high concentration AU intervention groups(P < 0.05). Compared with Ang Ⅱ intervention group, low, medium and high concentration of AU intervention group ROS, the production of collagen type Ⅰ, Ⅲ decreased significantly(P < 0.05). Conclusion AU can restrain by ROS-MAPK pathway Ang Ⅱ induction of CFs proliferation and collagen formation.
引文
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[13]赖银璇,王明蕊,杨海丽,等.柚皮素通过ROS/JNK/Bcl2通路抑制宫颈癌Hela细胞增殖和迁移[J].中药药理与临床,2018,34(1):40-43.
[14]李梦,钱海兵.ROS-MAPK信号通路与心肌纤维化的研究进展[J].长春中医药大学学报,2017,33(1):166-168.
[15]孟哲,李海禹,陶海龙.基于TGF-β1/Smad3信号通路探讨虾青素对心肌成纤维细胞Ⅰ、Ⅲ型胶原表达的影响[J].中国药理学通报,2018,34(6):841-845.
[16]韦立群,李清,甘嘉亮,等.迷迭香酸衍生物RAD-9通过PI3K/Akt和p38 MAPK信号通路诱导胃癌MGC-803细胞凋亡[J].中国药理学通报,2018,34(2):256-259.
[17]Dai H,Jia G,Lu M,et al.Astragaloside IV inhibits isoprenaline-induced cardiac fibrosis by targeting the reactive oxygen species/mitogen activated protein kinase signaling axis[J].Mol Med Rep,2017,15(4):1765-1770.
[18]辛博,陈力,万丽丽,等.野黄芩苷对血管紧张素Ⅱ诱导的新生大鼠心肌成纤维细胞的增殖及ERK1/2、p38MAPK信号通路的影响[J].中国药房,2018,29(5):629-633.
[19]胡哲夫,唐其柱,刘源,等.橙皮素对转化生长因子-β1诱导心肌成纤维细胞增殖的影响[J].疑难病杂志,2015,14(4):376-379.
[20]张羽飞,孟娜娜,李厚忠,等.柚皮苷对糖尿病大鼠心肌纤维化及STAT3磷酸化水平的影响[J].药物评价研究,2018,41(1):48-54.
[2]陈伟伟,高润霖,刘力生,等.《中国心血管病报告2015》概要[J].中国循环杂志,2016,31(6):521-528.
[3]董浩,韩旭晨,刘巍.抗心肌纤维化治疗的研究进展[J]现代医学,2017,45(10):1521-1524.
[4]马金,丁春华.心脏成纤维细胞与心肌纤维化[J].中华心血管病杂志,2014,42(3):269-272.
[5]韩曼飞,张刘强,李医明.天然桃叶珊瑚苷及其衍生物的化学结构和药理作用研究进展[J].中草药,2017,48(19):4105-4113.
[6]阮德功,马龙,许晖,等.杜仲翅果桃叶珊瑚甙抗氧化活性的研究[J].食品工业科技,2011,32(8):120-122.
[7]郑杰,刘端,赵肃清,等.杜仲叶桃叶珊瑚苷的酶法提取及其抑菌活性[J].中药材,2012,35(2):304-306.
[8]Xue HY,Gao GZ,Lin QY,et a1.Protective effects of aucubin on H202.Induced apoptosis in PCI2 cells[J].Phytother Res,2012,26(3):369-374.
[9]Lyu PY,Feng H,Huang WH,et al.Aucubin and its hydrolytic derivative attenuate activation of hepatic stellate cells via modulation of TGF-βstimulation[J].Environ Toxicol Pharmacol,2017,50(3):234-239.
[10]于瑞,王幼平,崔琳,等.心肌纤维化的发病机制及其研究进展[J].中国现代医生,2015,53(13):157-160.
[11]李乾,常亮,苏冬梅,等.粉防己碱对心肌成纤维细胞增殖、活化的影响[J].北京大学学报:医学版,2018,50(2):331-334.
[12]董文珠,曹晓倩,王璐,等.AngⅡ-ROS-PKC/Nrf2/HO-1通路调控LX-2细胞增殖的机制研究[J].浙江中医药大学学报,2018,42(4):319-325.
[13]赖银璇,王明蕊,杨海丽,等.柚皮素通过ROS/JNK/Bcl2通路抑制宫颈癌Hela细胞增殖和迁移[J].中药药理与临床,2018,34(1):40-43.
[14]李梦,钱海兵.ROS-MAPK信号通路与心肌纤维化的研究进展[J].长春中医药大学学报,2017,33(1):166-168.
[15]孟哲,李海禹,陶海龙.基于TGF-β1/Smad3信号通路探讨虾青素对心肌成纤维细胞Ⅰ、Ⅲ型胶原表达的影响[J].中国药理学通报,2018,34(6):841-845.
[16]韦立群,李清,甘嘉亮,等.迷迭香酸衍生物RAD-9通过PI3K/Akt和p38 MAPK信号通路诱导胃癌MGC-803细胞凋亡[J].中国药理学通报,2018,34(2):256-259.
[17]Dai H,Jia G,Lu M,et al.Astragaloside IV inhibits isoprenaline-induced cardiac fibrosis by targeting the reactive oxygen species/mitogen activated protein kinase signaling axis[J].Mol Med Rep,2017,15(4):1765-1770.
[18]辛博,陈力,万丽丽,等.野黄芩苷对血管紧张素Ⅱ诱导的新生大鼠心肌成纤维细胞的增殖及ERK1/2、p38MAPK信号通路的影响[J].中国药房,2018,29(5):629-633.
[19]胡哲夫,唐其柱,刘源,等.橙皮素对转化生长因子-β1诱导心肌成纤维细胞增殖的影响[J].疑难病杂志,2015,14(4):376-379.
[20]张羽飞,孟娜娜,李厚忠,等.柚皮苷对糖尿病大鼠心肌纤维化及STAT3磷酸化水平的影响[J].药物评价研究,2018,41(1):48-54.
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