葆生精改善少弱精子症模型大鼠生殖功能的实验研究

Protective effect of BaoShengJing on the reproductive function in rats with oligoasthenospermia

目的研究葆生精对奥硝唑所致少弱精子症模型大鼠生殖功能的保护作用。方法取34只雄性SD大鼠,将大鼠按体质量随机分为溶剂对照组(7只)、少弱精子症模型组(8只)、生理盐水对照模型组(9只)和葆生精治疗模型组(10只)。除溶剂对照组外,其他各组灌服奥硝唑400 mg/(kg·d)。从奥硝唑灌胃的第15天开始,葆生精治疗模型组加用葆生精514 mg/(kg·d),生理盐水模型组加用葆生精等体积的生理盐水,连续给药三周。观察各组大鼠附睾精子浓度、活力和运动参数等指标变化,睾丸作HE染色观察组织形态学改变,检测睾丸中抗氧化酶SOD、GSH-Px酶活性,抗凋亡蛋白Bcl-2和凋亡蛋白Caspase-3的表达变化。结果与溶剂对照组相比,少弱精子症模型组大鼠附睾精子密度明显降低[(16.92±1.31)×10~6/m L vs (31.62±4.44)×10~6/m L],活力也明显降低[(54.13±2.58)%vs (88.33±4.04)%],差异均有统计学意义(P均<0.01)。葆生精治疗模型组大鼠附睾的精子密度和活力比少弱精子症模型组明显增加,两组的密度分别为(26.34±3.16)×10~6/m L和(16.92±1.31)×10~6/m L,两组的活力分别为(78.4±2.43)%和(54.13±2.58)%,两组比较,P均<0.01,而生理盐水对照组与少弱精子症模型组无明显差异,各组中直线速率、曲线速率等精子运动参数也表现为相同趋势。少弱精子症模型组大鼠睾丸组织中部分生精细胞脱落,层次排列紊乱,生精上皮细胞变薄,管腔内精子数量明显减少;葆生精治疗组大鼠睾丸中虽然各级生精细胞排列略稀疏,但生精上皮形态基本恢复,各级生精细胞及管腔内精子数目都有明显增加。与溶剂对照组相比,少弱精子症模型组大鼠睾丸中SOD和GSH-Px的酶活力均明显降低,分别为[(10.71±1.34) U/mg vs (30.02±5.42) U/mg和(58.33±20.23) U/mg vs (175.00±27.9) U/mg,差异均有统计学意义(P均<0.01),葆生精治疗组两种酶活力均明显增加,分别为(34.30±8.24) U/mg和(220±55.41) U/mg。同时我们观察到与对照组相比,少弱精子症模型大鼠睾丸组织中抗凋亡蛋白Bcl-2的表达有所下降而凋亡蛋白Caspase-3的表达明显增加,葆生精治疗组Bcl-2表达增加而Caspase-3的表达下降。结论葆生精可通过提高少弱精子症模型大鼠的抗氧化能力,修复生殖系统的氧化损伤,抑制生精细胞凋亡,从而提高精子密度和活力,发挥对生殖系统的保护作用。

Objective To study th e protective effect of BaoShengJing on the reproductive function of rats with ornidazole-induced oligoasthenospermia(OAS). Methods Thirty-four male SD rats were randomly divided into the normal control group, the OAS model group, the OAS model group treated with normal saline(NS) and the OAS model group treated with BaoShengJing. OAS models were made in the latter three groups by intragastrical administration of ornidazole at 400 mg per kg of the body weight per day. From the 15 th day, additional BaoShengJing [514 mg/(kg·d)] was administrated to the rats in the group of BaoShengJing-treated OAS model, and equal NS to the rats in the group of NS-treated OAS model, for 3 successive weeks. After treatment, we detected the concentration, motility and kinematic parameters of the epididymal sperm,then examined the enzymatic activity of superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px), apoptosis-related protein B-cell lymphoma-2(bcl-2) and Caspase-3, and the histomorphological changes in testis. Results Compared with that in the normal control group, the concentration and motility of epididymal sperm in the group of OAS model were markedly decreased [(16.92 ±1.31) ×10~6/m L vs(31.62 ±4.44) ×106/m L;(54.13 ±2.58)% vs(88.33 ±4.04)%,P< 0.01], the sperm concentration and motility were increased to(26.34 ±3.16) ×10~6/mL and(78.4 ±2.43)%, respectively after treatment of BaoShengJing, while there was no significant change in the group of NS-treated OAS model. Testis tissues of rats in the group of OAS model showed deciduous spermatogenic cells, disorderly and loose arrangement of seminiferous tubules, thinned seminiferous epithelium and decreased sperm count, while testis tissues of rats in BaoShengJing-treated group exhibited ordered seminiferous epithelium and increased spermatogenic cells and sperm count, although remained loose arranged seminiferous tubules. Compared with that in the normal control group, the testicular enzymatic activities in the group of OAS model were markedly decreased [(10.71±1.34) U/mg vs(30.02±5.42) U/mg;(58.33±20.23) U/mg vs(175.00±27.9) U/mg,P< 0.01], the two enzymatic activities were increased to(34.30±8.24) U/mg 和(220±55.41) U/mg, respectively after treatment of BaoShengJing. Downregulation of bcl-2 and upregulation of Caspase-3 were detected in the group of OAS models compared with the normal control group, while BaoShengJing treatment reversed these destructive effects. Conclusion BaoShengJing can effectively improve sperm quality and protect the reproductive system of ornidazole-induced OAS rats by elevating antioxidant ability, reducing oxidative stress injury, inhibiting spermatogenic cell apoptosis.