龙胆苦苷对大鼠血管平滑肌细胞增殖、迁移的影响
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摘要
目的观察龙胆苦苷对血管平滑肌细胞(vascular smooth muscle cells,VSMC)增殖、迁移能力的影响,并探讨其可能的作用机制。方法体外分离培养大鼠血管平滑肌细胞,将血管平滑肌细胞随机分为对照组、龙胆苦苷(GPS)处理组、PF-3758309+龙胆苦苷组。采用噻唑蓝(MTT)法观察各组细胞增殖能力,计算细胞增殖抑制率;采用细胞划痕实验观察各组细胞迁移能力,计算细胞迁移率。采用Western blotting法检测各组细胞Pak4及MMP-2表达情况。结果与对照组相比,龙胆苦苷组细胞增殖率及迁移率明显降低(P<0.01),龙胆苦苷组细胞内Pak4、MMP-2表达水平明显降低(P<0.01);而与龙胆苦苷组相比,PF-3758309+龙胆苦苷组可明显增加细胞增殖率及迁移率(P<0.05),血管平滑肌细胞内MMP-2表达水平明显上调(P<0.05)。结论龙胆苦苷具有抑制血管平滑肌细胞增殖及迁移,其机制可能与抑制Pak4相关。
Objective To investigate the effects of gentiopicroside(GPS) on proliferation and migration of vascular smooth muscle cells.Methods Vascular smooth muscle cells(VSMC) were obtained from rat thoracic aorta,and then randomly divided into control group,GPS group and PF-3758309+GPS group.MTT was used to detected the effects of GPS on proliferation of VSMCs;The cell scratch test was used to detected the effects of GPS on cell migration; The indicator were detected by Western blot,which included the expression level of p-21 activated kinase-4(Pak4) and Metal matrix protein-2(MMP-2).Results Compared with control group,the proliferation inhibitory and migration rates of cells after treatment with GPS for 48 h were significantly decreased(P<0.01),and the expression of Pak4 and MMP-2 protein of cells after treatment with GPS for 48 h were significantly down-regulated(P<0.01).After adding PF-3758309,the proliferation inhibitory and migration rates were higher than these of GPS group,and the expression of MMP-2 was higher than that of the GPS group,and the differences were statistically significant(P<0.05).Conclusion The GPS can inhibit the proliferation and migration of vascular smooth muscle cells through inhibition of abnormal Pak4 expression.
Objective To investigate the effects of gentiopicroside(GPS) on proliferation and migration of vascular smooth muscle cells.Methods Vascular smooth muscle cells(VSMC) were obtained from rat thoracic aorta,and then randomly divided into control group,GPS group and PF-3758309+GPS group.MTT was used to detected the effects of GPS on proliferation of VSMCs;The cell scratch test was used to detected the effects of GPS on cell migration; The indicator were detected by Western blot,which included the expression level of p-21 activated kinase-4(Pak4) and Metal matrix protein-2(MMP-2).Results Compared with control group,the proliferation inhibitory and migration rates of cells after treatment with GPS for 48 h were significantly decreased(P<0.01),and the expression of Pak4 and MMP-2 protein of cells after treatment with GPS for 48 h were significantly down-regulated(P<0.01).After adding PF-3758309,the proliferation inhibitory and migration rates were higher than these of GPS group,and the expression of MMP-2 was higher than that of the GPS group,and the differences were statistically significant(P<0.05).Conclusion The GPS can inhibit the proliferation and migration of vascular smooth muscle cells through inhibition of abnormal Pak4 expression.
引文
[1]ROSAS-PERALTA M,BORRAYO-SNCHEZ G,MADRID-MILLER A,et al.Cardiovascular complications of hypertensive crisis[J].Rev Med Inst Mex Seguro Soc,2016,54(Suppl1):s67-s74.
[2]CONWAY B D,STAMOU S C,KOUCHOUKOS N T,et al.Improved clinical outcomes and survival following repair of acute type A aortic dissection in the current era[J].Interact Cardiovasc Thorac Surg,2014,19(6):971-976.
[3]CONRAD M F,CARVALHO S,ERGUL E,et al.Late aortic remodeling persists in the stented segment after endovascular repair of acute complicated type B aortic dissection[J].J Vasc Surg,2015,62(3):600-605.
[4]NIU Y T,ZHAO Y P,JIAO Y F,et al.Protective effect of gentiopicroside against dextran sodium sulfate induced colitis in mice[J].Int Immunopharmacol,2016(39):16-22.
[5]TANG X,YANG Q,YANG F,et al.Target profiling analyses of bile acids in the evaluation of hepatoprotective effect of gentiopicroside on ANIT-induced cholestatic liver injury in mice[J].J Ethnopharmacol,2016(194):63-71.
[6]SENOL F S,TUZUN C Y,TOKER G,et al.An in vitro perspective to cholinesterase inhibitory and antioxidant activity of five Gentiana species and Gentianella caucasea[J].Int JFood Sci Nutr,2012,63(7):802-812.
[7]WEI S,CHEN G,HE W,et al.Inhibitory effects of secoiridoids from the roots of Gentiana straminea on stimulusinduced superoxide generation,phosphorylation and translocation of cytosolic compounds to plasma membrane in human neutrophils[J].Phytother Res,2012,26(2):168-173.
[8]王戈,王宁,易蔚,等.龙胆苦苷对压力负荷致小鼠心肌肥厚的保护作用[J].中国动脉硬化杂志,2014,22(10):981-987.
[9]YANG Y,DUAN W,LIANG Z,et al.Curcumin attenuates endothelial cell oxidative stress injury through Notch signaling inhibition[J].Cell Signal,2013,25(3):615-629.
[10]SEGUCHI R,OHTAKE H,YOSHIMURA T,et al.Emergency Thoracic Endovascular Aortic Repair of Ruptured Kommerell's Diverticulum with an Acute Aortic Dissection[J].Kyobu Geka,2016,69(6):443-446.
[11]BARBETSEAS J,ALEXOPOULOS N,BRILI S,et al.Atherosclerosis of the aorta in patients with acute thoracic aortic dissection[J].Circ J,2008,72(11):1773-1776.
[12]NASTASIJEVI C'B,LAZAREVI C'-PATI T,DIMITRIJEVI C'-BRANKOVI S,et al.Inhibition of myeloperoxidase and antioxidative activity of Gentiana lutea extracts[J].J Pharm Biomed Anal,2012(66):191-196.
[13]乔慧莲,陈浩,陈文豪,等.龙胆苦甙后处理对心脏缺血再灌注损伤的防治作用[J].山东医药,2013,53(35):1-4.
[14]满枋霖,赵良智,孙新.p21活化激酶在肿瘤中的作用[J].中国生物化学与分子生物学报,2015,31(6):579-585.
[15]TYAGI N,MARIMUTHU S,BHARDWAJ A,et al.p-21 activated kinase 4(PAK4)maintains stem cell-like phenotypes in pancreatic cancer cells through activation of STAT3 signaling[J].Cancer Lett,2016,370(2):260-267.
[16]NEKRASOVA T,MINDEN A.Role for p21-activated kinase PAK4 in development of the mammalian heart[J].Transgenic Res,2012,21(4):797-811.
[2]CONWAY B D,STAMOU S C,KOUCHOUKOS N T,et al.Improved clinical outcomes and survival following repair of acute type A aortic dissection in the current era[J].Interact Cardiovasc Thorac Surg,2014,19(6):971-976.
[3]CONRAD M F,CARVALHO S,ERGUL E,et al.Late aortic remodeling persists in the stented segment after endovascular repair of acute complicated type B aortic dissection[J].J Vasc Surg,2015,62(3):600-605.
[4]NIU Y T,ZHAO Y P,JIAO Y F,et al.Protective effect of gentiopicroside against dextran sodium sulfate induced colitis in mice[J].Int Immunopharmacol,2016(39):16-22.
[5]TANG X,YANG Q,YANG F,et al.Target profiling analyses of bile acids in the evaluation of hepatoprotective effect of gentiopicroside on ANIT-induced cholestatic liver injury in mice[J].J Ethnopharmacol,2016(194):63-71.
[6]SENOL F S,TUZUN C Y,TOKER G,et al.An in vitro perspective to cholinesterase inhibitory and antioxidant activity of five Gentiana species and Gentianella caucasea[J].Int JFood Sci Nutr,2012,63(7):802-812.
[7]WEI S,CHEN G,HE W,et al.Inhibitory effects of secoiridoids from the roots of Gentiana straminea on stimulusinduced superoxide generation,phosphorylation and translocation of cytosolic compounds to plasma membrane in human neutrophils[J].Phytother Res,2012,26(2):168-173.
[8]王戈,王宁,易蔚,等.龙胆苦苷对压力负荷致小鼠心肌肥厚的保护作用[J].中国动脉硬化杂志,2014,22(10):981-987.
[9]YANG Y,DUAN W,LIANG Z,et al.Curcumin attenuates endothelial cell oxidative stress injury through Notch signaling inhibition[J].Cell Signal,2013,25(3):615-629.
[10]SEGUCHI R,OHTAKE H,YOSHIMURA T,et al.Emergency Thoracic Endovascular Aortic Repair of Ruptured Kommerell's Diverticulum with an Acute Aortic Dissection[J].Kyobu Geka,2016,69(6):443-446.
[11]BARBETSEAS J,ALEXOPOULOS N,BRILI S,et al.Atherosclerosis of the aorta in patients with acute thoracic aortic dissection[J].Circ J,2008,72(11):1773-1776.
[12]NASTASIJEVI C'B,LAZAREVI C'-PATI T,DIMITRIJEVI C'-BRANKOVI S,et al.Inhibition of myeloperoxidase and antioxidative activity of Gentiana lutea extracts[J].J Pharm Biomed Anal,2012(66):191-196.
[13]乔慧莲,陈浩,陈文豪,等.龙胆苦甙后处理对心脏缺血再灌注损伤的防治作用[J].山东医药,2013,53(35):1-4.
[14]满枋霖,赵良智,孙新.p21活化激酶在肿瘤中的作用[J].中国生物化学与分子生物学报,2015,31(6):579-585.
[15]TYAGI N,MARIMUTHU S,BHARDWAJ A,et al.p-21 activated kinase 4(PAK4)maintains stem cell-like phenotypes in pancreatic cancer cells through activation of STAT3 signaling[J].Cancer Lett,2016,370(2):260-267.
[16]NEKRASOVA T,MINDEN A.Role for p21-activated kinase PAK4 in development of the mammalian heart[J].Transgenic Res,2012,21(4):797-811.
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