Yan Jun-Feng, Zhao Jian-Feng, Zhang Gao-Yue, Huang Xiao-Jun, Chen Gang, Lü Bo-Dong
The Second School of Clinical Medicine, Zhejiang University of Traditional Chinese Medicine, Hangzhou, Zhejiang 310053, China.
Department of Urology, The Second Hospital Affiliated to Zhejiang University of Chinese Medicine, Hangzhou, Zhejiang 310005, China.
Zhonghua Nan Ke Xue. 2016 Aug;22(8):725-729.
To investigate the effect of salidroside on the expression of the connexin43 (Cx43) protein in the corpus cavernosum smooth muscle cells (CCSMCs) of hypoxic SD rats.
CCSMCs were cultured in vitro and identified by immunofluorescence staining. The cells were divided into six groups: normal control (21% O2), hypoxia (1% O2), hypoxia+salidroside (HS) 8 μg/ml,HS 16 μg/ml, HS 32 μg/ml, and HS 64 μg/ml, and cultured for 48 hours. Then the relative expression of Cx43 in different groups was detected by Western blot.
The in vitro cultured CCSMCs grew well and 90% of the cells showed positivity for α-SMA and desmin on immunohistochemistry. Salidroside ≤64 μg/ml produced no obvious toxicity on the CCSMCs. The expressions of Cx43 and phosphorylated proteins were dramatically increased in the hypoxia group as compared with the normal control (P<0.01 and P<0.05). The HS groups all showed significantly higher expression of Cx43 than the hypoxia group (P<0.01), but the phosphorylation rate of the Cx43 proteins was remarkably decreased (P<0.01).
Hypoxia increases the expression of Cx43 in the CCSMCs of SD rats. Salidroside ≤64 μg/ml cannot reverse the hypoxia-induced change but can reduce the dephosphorylation of Cx43 in CCSMCs. It is deduced that salidroside can protect CCSMCs by decreasing the phosphorylation of Cx43 and suppressing hypoxia-induced formation of the gap junction channel.
探讨红景天苷对缺氧SD大鼠阴茎海绵体平滑肌细胞（CCSMCs）中连接蛋白43（Cx43）蛋白表达的影响。
体外培养CCSMCs并通过免疫荧光染色进行鉴定。将细胞分为六组：正常对照组（21% O₂）、缺氧组（1% O₂）、缺氧+红景天苷（HS）8 μg/ml组、HS 16 μg/ml组、HS 32 μg/ml组和HS 64 μg/ml组，培养48小时。然后通过蛋白质免疫印迹法检测不同组中Cx43的相对表达。
体外培养的CCSMCs生长良好，免疫组化显示90%的细胞α-SMA和结蛋白呈阳性。红景天苷浓度≤64 μg/ml时对CCSMCs无明显毒性。与正常对照组相比，缺氧组Cx43及其磷酸化蛋白的表达显著增加（P<0.01和P<0.05）。HS各处理组Cx43的表达均显著高于缺氧组（P<0.01），但Cx43蛋白的磷酸化率显著降低（P<0.01）。
缺氧可增加SD大鼠CCSMCs中Cx43的表达。红景天苷浓度≤64 μg/ml不能逆转缺氧诱导的变化，但可减少CCSMCs中Cx43的去磷酸化。推测红景天苷可通过降低Cx43的磷酸化并抑制缺氧诱导的缝隙连接通道形成来保护CCSMCs。
