Chen Jian-Guo, Jiang Rui
Department of Urology, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan 646000, China.
Zhonghua Nan Ke Xue. 2019 Oct;25(10):874-882.
To investigate the improving effect of S1P3 silencing on the erectile function of spontaneous hypertension rats (SHR).
Five 12-week-old healthy male WKY rats were included in group A and another 15 12-week-old healthy male SHRs equally randomized into groups B1 (intracavernously injected with 20 μl S1P3 siRNA lentiviral vectors [2 ×108TU/ml]), B2 (intracavernously injected with 20 μl GFP lentiviral vectors [2 ×108TU/ml]), and C (control). At 1 week after transfection, the ratio of the maximum intracavernous pressure/mean arterial pressure (ICPmax/MAP) of the rats was measured, and the expressions of S1P3, ROCK1, ROCK2 and eNOS in the corpus cavernosal tissue were determined by immunohistochemistry, Western blot and RT-qPCR.
There were no statistically significant differences in the body weight and serum T level among the three groups of rats. Under 0 V, 3 V and 5 V electrical stimulation, the ratios of ICPmax/MAP were 0.16 ± 0.01, 0.55 ± 0.03 and 0.82 ± 0.02 in group A, 0.15 ± 0.01, 0.55 ± 0.01 and 0.79 ± 0.03 in group B1, 0.10 ± 0.00, 0.22 ± 0.01 and 0.43 ± 0.01 in group B2, and 0.11 ± 0.01, 0.22 ± 0.01 and 0.42 ± 0.02 in group C, significantly lower in the latter two than in the former two groups (P < 0.05). The protein expressions of S1P3, ROCK1 and ROCK2 in the corpus cavernosum were remarkably down-regulated while that of eNOS up-regulated in groups A and B1 compared with those in groups B2 and C (P < 0.05). Groups A and B1, in comparison with B2 and C, also showed markedly decreased mRNA expressions of S1P3 (0.72 ± 0.04 and 0.71 ± 0.07 vs 1.00 ± 0.06 and 1.00 ± 0.10, P < 0.05), ROCK1 (0.99 ± 0.05 and 1.08 ± 0.16 vs 1.85 ± 0.44 and 2.02 ± 0.38, P < 0.05) and ROCK2 (1.00 ± 0.03 and 1.08 ± 0.16 vs 2.16 ± 0.78 and 2.46 ± 0.69, P < 0.05), but increased eNOS (1.04 ± 0.15 and 0.81 ± 0.23 vs 0.32 ± 0.08 and 0.32 ± 0.04, P < 0.05).
S1P3 siRNA can improve the erectile function of SHRs by inhibiting the expression of the S1P3 gene and down-regulating the RhoA/Rho kinase signaling pathway in the corpus cavernosum.
探讨沉默S1P3对自发性高血压大鼠(SHR)勃起功能的改善作用。
将5只12周龄健康雄性WKY大鼠纳入A组,另将15只12周龄健康雄性SHR平均随机分为B1组(海绵体内注射20 μl S1P3 siRNA慢病毒载体[2×108 TU/ml])、B2组(海绵体内注射20 μl GFP慢病毒载体[2×108 TU/ml])和C组(对照组)。转染1周后,测量大鼠海绵体内最大压力/平均动脉压(ICPmax/MAP)比值,采用免疫组织化学、蛋白质印迹法和RT-qPCR检测海绵体组织中S1P3、ROCK1、ROCK2和eNOS的表达。
三组大鼠体重和血清睾酮水平差异无统计学意义。在0 V、3 V和5 V电刺激下,A组的ICPmax/MAP比值分别为0.16±0.01、0.55±0.03和0.82±0.02,B1组分别为0.15±0.01、0.55±0.01和0.79±0.03,B2组分别为0.10±0.00、0.22±0.01和0.43±0.01, C组分别为0.11±0.01、0.22±0.01和0.42±0.02,后两组明显低于前两组(P<0.05)。与B2组和C组相比,A组和B1组海绵体中S1P3、ROCK1和ROCK2的蛋白表达明显下调,而eNOS的蛋白表达上调(P<0.05)。与B2组和C组相比,A组和B1组S1P3(0.72±0.04和0.71±0.07 vs 1.00±0.06和1.00±0.10,P<0.05)、ROCK1(0.99±0.05和1.08±0.16 vs 1.85±0.44和2.02±0.38,P<