CAJ | 학술논문

目的:观察丹参酮ⅡA( TanⅡA)对体外培养的大鼠血管平滑肌细胞( VSMC)增殖与迁移的影响. 方法:采用CCK8法筛选VSMC增殖的最适胎牛血清(FBS)浓度,然后将VSMC分为3组:(1)对照组:VSMC加入5%FBS培养液;(2)增殖组:VSMC加入30%FBS培养液;( 3 ) TanⅡA干预组:①10 mg/L TanⅡA+30%FBS;②20 mg/L TanⅡA +30%FBS;③40 mg/L TanⅡA+30%FBS,分别培养24、48、72 h后检测细胞增殖情况;伤口愈合实验观察VSMC迁移指数;免疫细胞化学法检测细胞血小板生长因子( PDGF)、碱性成纤维细胞生长因子( bFGF)的表达变化;应用Western blot法观察细胞凋亡相关蛋白Bcl-2、Bax及金属基质蛋白酶2(MMP2)、金属基质蛋白酶9(MMP9)的表达情况. 结果:与增殖组相比,TanⅡA干预后细胞增殖明显缓慢,其抑制作用呈时间、剂量依赖性,20 mg/L TanⅡA干预48 h对VSMC增殖的抑制作用比较显著,且无明显细胞损伤. 伤口愈合实验显示TanⅡA干预后VSMC的迁移指数明显低于增殖组(P<0. 05);免疫细胞化学结果显示,与增殖组相比,TanⅡA干预后PDGF表达减少,差异具有统计学意义(P<0. 05),各组间bFGF表达差异无统计学意义;Western blot结果显示,与增殖组比较,TanⅡA干预组随药物浓度的提高,Bcl-2、MMP2、MMP9表达水平降低,Bax提高,差异具有统计学意义(P<0. 05). 结论:20 mg/L TanⅡA对高浓度FBS诱导的VSMC增殖与迁移的抑制作用较为明显,且无明显细胞损伤,其抑制作用可能与调节Bcl-2、Bax表达,阻断PDGF、MMP2、MMP9生长因子活化有关.
목적:관찰단삼동ⅡA( TanⅡA)대체외배양적대서혈관평활기세포( VSMC)증식여천이적영향. 방법:채용CCK8법사선VSMC증식적최괄태우혈청(FBS)농도,연후장VSMC분위3조:(1)대조조:VSMC가입5%FBS배양액;(2)증식조:VSMC가입30%FBS배양액;( 3 ) TanⅡA간예조:①10 mg/L TanⅡA+30%FBS;②20 mg/L TanⅡA +30%FBS;③40 mg/L TanⅡA+30%FBS,분별배양24、48、72 h후검측세포증식정황;상구유합실험관찰VSMC천이지수;면역세포화학법검측세포혈소판생장인자( PDGF)、감성성섬유세포생장인자( bFGF)적표체변화;응용Western blot법관찰세포조망상관단백Bcl-2、Bax급금속기질단백매2(MMP2)、금속기질단백매9(MMP9)적표체정황. 결과:여증식조상비,TanⅡA간예후세포증식명현완만,기억제작용정시간、제량의뢰성,20 mg/L TanⅡA간예48 h대VSMC증식적억제작용비교현저,차무명현세포손상. 상구유합실험현시TanⅡA간예후VSMC적천이지수명현저우증식조(P<0. 05);면역세포화학결과현시,여증식조상비,TanⅡA간예후PDGF표체감소,차이구유통계학의의(P<0. 05),각조간bFGF표체차이무통계학의의;Western blot결과현시,여증식조비교,TanⅡA간예조수약물농도적제고,Bcl-2、MMP2、MMP9표체수평강저,Bax제고,차이구유통계학의의(P<0. 05). 결론:20 mg/L TanⅡA대고농도FBS유도적VSMC증식여천이적억제작용교위명현,차무명현세포손상,기억제작용가능여조절Bcl-2、Bax표체,조단PDGF、MMP2、MMP9생장인자활화유관.
Objective:To investigate the effects of tanshinoneⅡA ( TanⅡA) on proliferation and migration of rat vascular smooth muscle cells ( VSMC) cultured in vitro. Methods:Screening optimal fetal bovine serum ( FBS) concentration on VSMC prolif-eration by CCK8, then VSMC were divided into three groups:(1) Control group:VSMC were added to 5% FBS culture medium;(2) Proliferation group:VSMC were added to 30% FBS culture medium; ( 3 ) TanⅡA intervention group:①10 mg/L TanⅡA+30%FBS;②20 mg/L TanⅡA+30%FBS; ③40 mg/L TanⅡA+30%FBS. After VSMC cultured 24, 48, 72 hours respectively, cell proliferation was detected. Wound healing experiment was to observe VSMC migration index. The expression of PDGF and bFGF were examined immunocytochemically. Western blot was used to observe the Bcl -2, Bax, MMP2 and MMP9 expression. Results:As compared with the proliferation groups,the cell proliferation of induced by TanⅡA was significantly slower and the inhibition depends on dose and time. VSMC proliferation inhibition was more significant which was induced by 20 mg/L TanⅡA intervention 48 h and no significant cell damage. Wound healing experiment showed that VSMC migration index was significantly lower than proliferation group after the intervention of TanⅡA (P<0. 05). Immunocytochemical staining showed that compared with the proliferation group, ex-pression of PDGF had reduced after the TanⅡA intervention, the difference was statistically significant (P<0. 05). Each group had no significant difference in the expression of bFGF. Western blot showed that with the increasing of drug concentration, the Bcl-2, MMP2, MMP9 expression levels of TanⅡA intervention group was lower, Bax was higher which was compared with proliferation group, the difference was statistically significant (P<0. 05). Conclusion:The inhibition of 20 mg/L TanⅡA on VSMC proliferation and migration which were induced by high concentration of FBS was more obvious and no significant cell damage, its inhibition may related to regulate Bcl-2, Bax expression and block activation of PDGF, MMP2, MMP9.