益肾通络方对精索静脉曲张模型大鼠生精细胞凋亡、caspase-3蛋白表达的影响

Effect of Yishen Tongluo Prescription on Apoptosis of Spermatogenic Cells and Modelcaspase-3 Protein of Model Rats with Varicocele

目的:观察益肾通络方对精索静脉曲张模型大鼠睾丸质量、生精细胞、caspase-3蛋白表达、睾丸生精细胞凋亡指数(apoptosis index,AI)的影响。方法:取清洁级雄性SD大鼠40只随机均分为5组:假手术组、精索静脉曲张模型组、益肾通络方低剂量组、益肾通络方中剂量组、益肾通络方高剂量组。模型建立4周后,假手术组及模型组均按15 m L·(kg·d)^(-1)给予去离子水,益肾通络方低剂量组、益肾通络方中剂量组、益肾通络方高剂量组分别给予益肾通络方颗粒剂混悬液1.5g·(kg·d)^(-1)、3.0 g·(kg·d)^(-1)、6.0 g·(kg·d)^(-1),灌胃,1次·d-1,持续60 d。60 d后5组大鼠均脱臼处死并分别取双侧睾丸,电子天平称取睾丸质量,并检测caspase-3在睾丸组织中的表达,生精细胞的凋亡并计算AI。结果:(1)睾丸质量的变化:5组大鼠左右两侧睾丸质量:假手术组为[(1.88±0.19)、(1.95±0.20)g]、精索静脉曲张模型组为[(0.72±0.23)、(1.31±0.34)g]、益肾通络方低剂量组[(0.73±0.28)、(1.39±0.31)g]、益肾通络方中剂量组为[(0.78±0.25)、(1.42±0.22)g]、益肾通络方高剂量组为[(1.08±0.28)、(1.64±0.33)g],假手术组、精索静脉曲张模型组与益肾通络方高剂量组大鼠两侧睾丸质量比较,差异具有统计学意义(P<0.05);益肾通络方低剂量组、益肾通络方中剂量组与精索静脉曲张模型组比较,差异无统计学意义(P>0.05)。(2)益肾通络方各剂量组大鼠的生精细胞caspase-3蛋白表达低于精索静脉曲张模型组,高于假手术组;益肾通络方低剂量组大鼠生精细胞caspase-3蛋白表达高于中剂量组,益肾通络方中剂量组生精细胞caspase-3蛋白表达高于高剂量组,差异均有统计学意义(P<0.05)。(3)假手术组AI评分最低,益肾通络方干预组AI评分低于精索静脉曲张模型组;益肾通络方干预组组内比较,高剂量组低于中剂量组,中剂量组低于低剂量组,差异均有统计学意义(P<0.05)。结论:精索静脉曲张模型大鼠睾丸质量减轻、生精细胞caspase-3蛋白表达升高、AI增加,这可能是影响生育力的机制之一。单侧精索静脉曲张引起睾丸损害是双侧的,但右侧比左侧损害轻。益肾通络方能够降低生精细胞caspase-3蛋白表达,减少精索静脉曲张大鼠睾丸生精细胞凋亡,对睾丸生精功能具有保护作用,进而可提高睾丸的生殖能力。

Objective： To observe the effects of Yishen Tongluo Prescription on testicular quality,spermatogenic cells,caspase-3 protein expression and testicular spermatogenic apoptosis index（ AI） in model rats with varicocele. Methods： Forty male SpragueDawley rats were randomly divided into 5 groups： sham operation group,varicocele model group,Yishen Tongluo Prescription lowdose group,Yishen Tongluo Prescription middle-dose group,Yishen Tongluo Prescription high-dose group. Four weeks after the model was established,the sham operation group and the model group were given deionized water at 15 m L ·（kg·d）-1,the Yishen Tongluo Prescription low-dose group,the Yishen Tongluo Prescription middle dose group,and Yishentong Prescription highdose group of the compound was given Yilu Tongluo Prescription granule suspension 1. 5 g·（kg·d）-1,3. 0 g·（kg·d）-1,6. 0 g·（kg·d）-1,respectively,by gavaging,1 time·d-1,and that lasted for 60 days. After 60 days,the rats in each group were killed by dislocation and the testes were taken from both sides. The quality of the testes was weighed and the expression of caspase-3 in testicular tissues was detected. The apoptosis of spermatogenic cells was calculated and AI was calculated. Results：① Changes in testicular mass： testicular mass in the left and right sides of the 5 groups： the sham operation group [（ 1. 88 ± 0. 19）,（ 1. 95 ±0. 20 g） ]; the varicocele model group [（ 0. 72 ± 0. 23） ],（ 1. 31 ± 0. 34 g） ]; the Yishen Tongluo Prescription low-dose group（ 0. 73 ± 0. 28）,（ 1. 39 ± 0. 31 g） ],the Yishen Tongluo Prescription middle-dose group [[0. 78 ± 0. 25）,（ 1. 42 ± 0. 22 g） ],Yishen Tongluo Prescription high-dose group [（ 1. 08 ± 0. 28）,（ 1. 64 ± 0. 33 g） ]. The differences between the sham operation group,varicocele model group and Yishen Tongluo high-dose group rats in testicular quality were statistically significant（ P〈0. 05）. The differences between the quality of testis on both sides of the rats in the low-dose group of Yishen Tongluo Prescription,the middle dose group of Yishen Tongluo Prescription and the varicocele model group were not statistically significant（ P〈0. 05）.② The expression of caspase-3 protein in the spermatogenic cells of the rats of Yishen Tongluo Prescription was significantly lower than that of the varicocele model group and that was middle-dose group higher than that of the sham operation group; the spermatogenic cells caspase-3 in the Yishen Tongluo Prescription low-dose group was middle-dose group higher that in the Yishen Tongluo Prescription middle-dose group. The expression of caspase-3 protein in the Yishen Tongluo Prescription middle-dose group was middle-dose group higher than that in the high dose group（ P〈0. 05）.③ The sham operation group had the lowest AI score,the Yishen Tongluo Prescription intervention group had significantly lower AI score than that of the varicocele model group; the Yishen Tongluo Prescription intervention group was significantly lower than the middle dose group and the middle dose group was lower.In the low-dose group,the difference was statistically significant（ P〈0. 05）. Conclusion： The quality of testis in rats with varicocele model is reduced,the expression of caspase-3 protein in spermatogenic cells is increased,and AI is increased,which may be one of the mechanisms affecting fertility. Testicular damage caused by unilateral varicocele is bilateral,but the right side is lighter than the left side. Yishen Tongluo Prescription can reduce the expression of caspase-3 protein in spermatogenic cells,reduce the apoptosis of testicular spermatogenic cells in rats with varicocele,and protect the testicular spermatogenesis,which can improve the reproductive capacity of testes.