活血化瘀法对低氧所致阴茎海绵体平滑肌细胞表型转化的影响

Effect of activating blood circulation therapy on the phenotypic transformation of corpus cavernous smooth muscle cells induced by hypoxia

目的探讨活血化瘀法对低氧所致阴茎海绵体平滑肌细胞(CCSMCs)表型转化的影响。方法选择10只雄性SD大鼠为研究对象,动物实验设血府逐瘀汤灌胃组和空白对照组。血府逐瘀汤含药血清的制备;体外培养SD大鼠CCSMCs,分别采用常氧(21%O 2)、低氧(1%O 2)、低氧及含药血清干预处理,即为常氧组、低氧组和血府逐瘀汤干预组,分别培养48h;免疫组化法鉴定细胞;Western印迹测定α-平滑肌肌动蛋白(α-SMA)和I型胶原蛋白(Collagen I)相对表达。结果体外培养CCSMC生长良好,抗α-平滑肌肌动蛋白(α-SMA)和抗Desmin抗体免疫组化染色阳性,CCSMC所占比例达90%以上。MTT法结果显示,血府逐瘀汤含药血清浓度<40%时,对CCSMC无明显毒性;而50%血府逐瘀汤含药血清对CCSMC有一定的抑制作用。未加含药血清干预时,与常氧组相比,低氧组Collagen I表达显著增加,α-SMA表达显著下降;与低氧不加含药血清组相比,40%含药血清组Collagen I表达相对下降,α-SMA表达显著增加。结论低氧促使大鼠CCSMCs表达α-SMA增加和Collagen I下降,40%血府逐瘀汤含药血清则使α-SMA表达下降和Collagen I表达下降,推测血府逐瘀汤在低氧所致CCSMCs发生表型转化的过程中起到抑制作用。

Objective To investigate the effect of XueFuZhuYu Decoction(XFZYD)on the phenotypic transformation of corpus cavernous smooth muscle cells induced by hypoxia.Methods Ten male SD rats were selected as the study subjects and were randomly divided into the XFZYD group and a blank control group.XFZYD containing serum was preparaed.CCSMCs were cultured in vitro and identified by immunofluorescence staining.In vitro,the cells were divided into normal control(21%O 2),hypoxia(1%O 2)and hypoxia+drug-containing serum,all cultured for 48 hours.The relative expression ofα-SMA and Collagen I in different groups was detected by western blot.Results CCSMCs which were cultured in vitro grew well and 90%of the cells showed positivity forα-SMA and Desmin on immunohistochemistry.XFZYD containing serum<40%produced no obvious toxicity on the CCSMCs.50%XFZYD containing serum had a certain inhibitory effect on CCSMCs.The expression ofα-SMA was remarkably decreased and Collagen I was dramatically increased in the hypoxia group as compared with the normal control.The 40%XFZYD containing serum group showed significantly higher expression ofα-SMA than the hypoxia group,but the expression of Collagen I was relatively decreased.Conclusions Hypoxia increases the expression of Collagen I and decreases the expression ofα-SMA in the CCSMCs.The 40%XFZYD containing serum increases the expression ofα-SMA and decreases the expression of Collagen I in the CCSMCs.It is speculated that XFZYD can inhibit the phenotypic transformation of CCSMCs induced by hypoxia.