| 刘冰,崔云,杜宝昕,等.通精灵对精索静脉曲张模型大鼠生精细胞凋亡及凋亡因子Cyt C、caspase-9的影响[J].浙江中医药大学学报,2019,43(1):26-33. |
| 通精灵对精索静脉曲张模型大鼠生精细胞凋亡及凋亡因子Cyt C、caspase-9的影响 |
| Effect of Tongjingling on Spermatogenic Cell Apoptosis and Apoptosis Factors Cyt C and Caspase-9 in Rats with Varicocele |
| DOI：10.16466/j.issn1005-5509.2019.01.005 |
| 中文关键词: 通精灵 精索静脉曲张 男性不育 生精细胞凋亡 细胞色素C caspase-9 |
| 英文关键词: Tongjingling varicocele male infertility spermatogenic cell apoptosis Cyt C caspase-9 |
| 基金项目:国家自然科学基金项目(81673985)；浙江省自然科学基金(LY16H270005) |
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| 中文摘要: |
| [目的]探讨通精灵对精索静脉曲张模型大鼠睾丸组织的保护作用及与凋亡因子细胞色素C(cytochrome C，Cyt C)、半胱氨酸蛋白酶-9(cysteine aspartic acid specific protease-9，caspase-9)表达的关系。[方法]72只SPF级Wistar大鼠，随机分成假手术组、模型组、通精灵低、中、高剂量组及左卡尼汀组。采用左肾静脉部分结扎法联合“夹尾激怒法”建立实验性精索静脉曲张合并中医肝气郁结证候复合大鼠模型。模型建立后，通精灵低、中、高剂量组分别给予浓度为1.6、3.2、6.4g·mL-1通精灵生药水煎液灌胃，假手术组和模型组予以0.9%氯化钠溶液灌胃，左卡尼汀组则予0.021g·mL-1左卡尼汀口服液灌胃，以上各组灌胃体积均为1mL/100g体质量，1次/d，持续8周。以HE染色观察睾丸组织结构；TUNEL染色和Annexin V-FITC法检测生精细胞凋亡情况；Western blot检测睾丸组织中Cyt C、caspase-9的表达。[结果]模型组中睾丸组织发生明显病理损伤，而通精灵各剂量组及左卡尼汀组中睾丸的病理损伤有不同程度改善。模型组中睾丸生精细胞凋亡率明显高于假手术组，差异有统计学意义(P＜0.05)，通精灵中、高剂量组及左卡尼汀剂量组中生精细胞凋亡率明显低于模型组，差异有统计学意义(P＜0.05或 P＜0.01)。Western blot提示，模型组大鼠睾丸组织中Cyt C、caspase-9蛋白表达显著高于假手术组(P＜0.01)；通精灵中、高剂量组及左卡尼汀组中Cyt C、caspase-9表达低于模型组(P＜0.05或P＜0.01)；高剂量组中Cyt C、caspase-9蛋白表达明显低于左卡尼汀组(P＜0.05，P＜0.01)。[结论]通精灵可以有效改善精索静脉曲张大鼠睾丸组织损伤，降低Cyt C和caspase-9的表达，从而调控生精细胞的凋亡。这可能是通精灵改善精索静脉曲张患者生育能力的机制之一。 |
| 英文摘要: |
| [Objective] To explore the protective effect of Tongjingling(TJL) on testicular tissue in rats with varicocele and its relationship with the expression of apoptosis factors cytochrome C(Cyt C) and cysteine aspartic acid specific protease-9(caspase-9).[Methods] Seventy two male SPF Wistar rats were randomly divided into six groups of equal number: sham operation group, model group, low-dose TJL, middle-dose TJL and high-dose TJL group and L-carnitine group. The composite rat model of experimental varicocele(EVC) with TCM syndrome of stagnation of liver Qi was induced by partial ligation of the left renal vein of rats and the method of clamping-tail. After establishment of varicocele model of rats, low-dose TJL, middle-dose TJL and high-dose TJL groups were given decoction with different concentrations of crude drugs(1.6g·mL-1, 3.2g·mL-1, 6.4g·mL-1). Rats in sham operation group and model group were given 0.9% saline. Rats in L-carnitine group were given L-carnitine oral liquid(0.021g·mL-1). All the above groups were gavaged according to 1mL/100g body weight, once a day for 8 weeks. The histological structure of the testis was observed by HE staining. TUNEL staining and Annexin V-FITC were used to detect the apoptosis of spermatogenic cells. The expression of Cyt C and caspase-9 in testicular tissue was detected by Western blot.[Results] In model group, the testicular tissue had obvious pathological damage, while the pathological damage of testicular tissue was improved in TJL groups and L-carnitine group. The apoptosis of testis spermatogonia in model group was significantly higher than that in sham operation group(P＜0.05). The apoptosis of the spermatogenic cells in middle-dose, high-dose TJL groups and L-carnitine group was significantly lower than that in model group(P＜0.05, P＜0.01). Compared with sham operation group, the expression of Cyt C and caspase-9 in testicular tissue of model group increased significantly(P＜0.01). Compared with model group, the expression of Cyt C and caspase-9 decreased with different degrees in middle-dose, high-dose TJL groups and L-carnitine group(P＜0.05, P＜0.01). Compared with L-carnitine group, the expression of Cyt C and caspase-9 protein in high-dose TJL group was significantly lower(P＜0.05, P＜0.01).[Conclusion] TJL can effectively improve testicular tissue damage in rats with varicocele, reduce the expression of Cyt C and caspase-9, and regulate the apoptosis of spermatogenic cells. Spermatogenic cells in rats with varicocele. This may be one of the molecular mechanism for improving the fertility of infertile patients with varicocele. |
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