津力达对高糖环境下大鼠阴茎海绵体平滑肌细胞表型转化的影响

Effect of Jinlida on Phenotypic Conversion of Corpus Cavernosum Smooth Muscle Cells in Rats Under High Glucose Conditions

目的探讨津力达颗粒对大鼠阴茎海绵体平滑肌细胞(corpus cavernosum smooth muscle cells,CCSMCs)表型转化的影响及机制。方法将原代SD大鼠CCSMCc进行传代培养,实验分为正糖组(NG组),高糖组(HG组),高渗组(HC组),高糖+PD98059组(HG+PD组),高糖+低、中、高浓度津力达组(HG+LJLD组、HG+MJLD组、HG+HJLD组),分别于0、24、48 h在高倍镜下观察细胞形态变化。Western blot检测各组α细胞平滑肌肌动蛋白(alpha smooth muscle actin,α-SMA)、碱性调宁蛋白1(calponin 1)和骨桥蛋白(osteopontin,OPN)以及信号通路蛋白细胞外信号调节激酶1/2 (extracellular signal-regulated kinases 1/2,ERK1/2),磷酸化细胞外信号调节激酶1/2 (phosphorylated extracellular signaling regulates kinases 1/2,p-ERK1/2)的表达。结果与NG组比较,HG组α-SMA、calponin 1表达均显著减少(P <0. 05); OPN表达显著增多(P <0. 05),p-ERK1/2表达增多(P <0. 01),HC组无上述改变。加入ERK抑制剂后p-ERK1/2表达减低(P <0. 05),α-SMA、calponin 1表达回升(P <0. 05); OPN表达减少(P <0. 05)。高糖加入津力达干预后,与HG组比较,HG+LJLD组、HG+MJLD组、HG+HJLD组calponin 1表达增多,p-ERK1/2表达减少(P均<0. 05),津力达浓度越大,逆转越明显; HG+HJLD组OPN表达显著减少(P <0. 05)。结论高糖可能通过上调ERK1/2的磷酸化进而诱导CCSMCs从收缩型向合成型转化;津力达可能通过抑制ERK1/2的磷酸化,改善高糖诱导的上述变化。

Objective To investigate the effect and mechanism of Jinlida(JLD)on the phenotypic changes of SD rats’corpus cavernosum smooth muscle cells(CCSMCs).Methods The primary SD rat CCSMCc was subcultured.The experiments were divided into normal glucose group(NG group),high glucose group(HG group),hyperosmotic group(HC group),high glucose+PD98059 group(HG+PD group)and high glucose plus low,medium,high concentrations of JLD groups(HG+LJLD group,HG+MJLD group,HG+HJLD group).The morphological changes of the cells were observed under high magnification at0,24 and 48 hours.The expressions of alpha smooth muscle actin(α-SMA),calponin 1,osteopontin(OPN)and extracellular signal-regulated kinases 1/2(ERK1/2),phosphorylated extracellular signaling regulates kinases 1/2(p-ERK1/2)were detected by western blot.Results Compared with NG group,the expressions ofα-SMA and calponin 1 in HG group were significantly decreased(P<0.05),the expression of OPN was significantly increased(P<0.05),and the expression of p-ERK1/2 was increased(P<0.01).There were no such changes in HC group.After the addition of ERK inhibitors,the expression of p-ERK1/2 was reduced,and changes inα-SMA,calponin 1,and OPN were recovered.After high-glucose was added to JLD intervention,the expressions of calponin 1,p-ERK1/2 in HG+LJLD group,HG+MJLD group and HG+HJLD group were reversed(all P<0.05).The greater the concentration of JLD,the more obvious the reversal.The expression of OPN in HG+HJLD group was significantly decreased(P<0.05).Conclusion High glucose may induce the increase of CCSMCs from contractile to synthetic by up-regulating ERK1/2 phosphorylation,JLD may improve the above-mentioned changes by high glucose through inhibiting ERK1/2 phosphorylation.