摘要
目的探讨百里醌作用于PTEN/AKT信号通路抑制大鼠平滑肌细胞增殖和迁移的影响,进一步深入了解其可能的作用机制.方法选取250~300g SD雄性大鼠分离并培养颈动脉平滑肌细胞,按照干预措施可分为空白对照组、低剂量百里醌组(1mg/L)、中剂量百里醌组(10mg/L)和高剂量百里醌组(100mg/L),百里醌各剂量组细胞用相应剂量的百里醌组进行干预.划痕实验观察平滑肌细胞迁移情况,MTT法、α-SMA和PCNA免疫组织化学染色检测颈动脉平滑肌细胞增殖情况,TUNEL染色检测颈动脉平滑肌细胞的凋亡情况,Western blot检测PTEN、P-AKT、AKT、Bax和bcl-2蛋白的表达量.结果与空白对照组比较,百里醌各剂量组迁移面积减小,OD值降低,α-SMA和PCNA表达阳性率降低,PTEN和Bax蛋白的表达量升高,P-AKT、AKT和Bcl-2蛋白的表达量降低,以上结果差异均有统计学意义(P<0.05).百里醌各剂量之间比较,随着剂量增大,迁移面积减小,OD值降低,α-SMA和PCNA表达阳性率降低,PTEN和Bax蛋白的表达量升高,P-AKT、AKT和Bcl-2蛋白的表达量降低,以上结果差异均有统计学意义(P<0.05).与其他组比较,高剂量百里醌组细胞凋亡率明显升高(P<0.05).结论百里醌能够抑制平滑肌细胞增殖和迁移,其作用机制与PTEN/AKT信号通路有关.
Objective To investigate the effect of thymoquinone on the proliferation and migration of rat smooth muscle cells(SMCs)induced by PTEN/AKT signaling pathway,and to further understand its possible mechanism.Methods SMCs were isolated and cultured from 250—300gSD male rats.According to the intervention measures,they were divided into blank control group,low dose thymol group(1 mg/L),middle dose thymol group(1Omg/L)and high dose thymol group(lOOmg/L).The cells of each dose group were interfered with the corresponding dose of thymoquinone group.The migration of smooth muscle cells was observed by scratch test,the proliferation of carotid smooth muscle cells was detected by MTT assay,α—SMA and PCNA histochemical staining,the apoptosis of carotid smooth muscle cells was detected by TUNEL staining,and the apoptosis of carotid smooth muscle cells was detected by Wester staining.N blot was used to detect the expression of PTEN,P—AKT,Bax andBcl—2 proteins.Results Compared with the blank control group,the migration area,the OD value,the positive rate ofα—SMA and PCNA expression,the expression of P-AKT,AKT and Bel—2 protein in each dose group were decreased,while the expression rate of PTEN and Bax protein were increased.There was significant difference between the above results(P<0.05).With the increase of the dose,the migration area was decreased,the OD value was decreased,the positive rate ofα—SMA and PCNA expression were decreased,the expression of PTEN and Bax protein were increased,and the expression of P—AKT,AKT and Bel—2 protein were decreased.It was statistically significant in three groups(P<0.05).Compared with other groups,the apoptosis rate of high dose thymol group was significantly higher than other groups(P<0.05).Conclusion Thymoquinone can inhibit the proliferation and migration of smooth muscle cells,which may be related to PTEN/AKT signaling pathway.
出处
《浙江临床医学》
2019年第12期1606-1608,1615,共4页
Zhejiang Clinical Medical Journal
基金
浙江省温州市科技计划经费自筹项目(Y20180487)。
