续断种子方对小鼠精子发生发育及c-kit蛋白表达的影响

Effects of Xuduan Zhongzi Formula on Generating and Developing Process of Mice Sperm and Protein Expression of c-kit

目的观察续断种子方对无精子症模型小鼠睾丸精子发生发育及c-kit蛋白表达的影响,探讨其相关作用机制。方法白消安和环磷酰胺腹腔注射建立无精子症小鼠模型。实验小鼠按随机数字表法分为对照组、模型组、麒麟丸组和续断种子方高、中、低剂量组。各治疗组予相应药物灌胃,对照组和模型组小鼠予等体积生理盐水,每日1次,连续8周。HE染色检测小鼠附睾精子质量,电镜观察睾丸组织显微、超微结构,免疫组化法检测睾丸组织c-kit蛋白表达。结果模型组偶见散在精子,睾丸生精上皮不同程度破坏,精原细胞、精母细胞线粒体空化明显,部分线粒体嵴消失,核膜局部破损,c-kit免疫组化结果显示棕黄色阳性表达的细胞散在;续断种子方高剂量组、麒麟丸组、对照组小鼠附睾精子浓度较高,睾丸生精上皮见精子、精子细胞、精母细胞,线粒体空化少,可见丰富的高尔基体、内质网结构,c-kit免疫组化结果显示棕黄色阳性表达的细胞灰度大。结论续断种子方可有效促进睾丸生精上皮的精子发生发育,增加精子数量,达到健脾益肾、活血生精之效。

Objective To observe the effects of Xuduan Zhongzi Formula on c-kit during the generating and developing processes of sperm of azoospermia mouse models. Methods Busulfan and cyclophosphamide intraperitoneal injections were used to establish azoospermia mouse models. Model mice were randomly divided into control group, model group, Qilin Pill group, high, medium and low dosage of Xuduan Zhongzi Formula groups through random number table method. Each treatment group was given relevant medicine for gavage, while control group and model group were given normal saline with the same volume for gavage, once a day for 8 weeks. HE dyeing was used to detect semen quality of epididymis;electron microscopy was used to observe ultrastructure of testicular tissue;immunohistochemistry was used to detect the protein expression of c-kit. Results Scattered sperms were occasionally seen in the model group. Testicular seminiferous epithelium was damaged to varying degrees. Obvious cavitation could be seen at the mitochondrias of spermatogonia and spermatocytes. Some of the mitochondria ridges disappeared, and some of the nuclear membrane was damaged. Scattered c-kit brown positive immunohistochemical expression cells were detected. Compared with the model group, a higher concentration of sperms were tested at the control group, Qilin Pill group, high, medium and low dosage of Xuduan Zhongzi Formula groups. Sperm, sperm cells and spermatocytes were seen at testicular seminiferous epithelium, and less cavitation was seen at the mitochondrias. Extensive Golgi apparatus and endoplasmic reticulum structure were detected. Greater cell brilliance of c-kit brown positive immunohistochemical expression cells was detected. Conclusion Xuduan Zhongzi Formula can effectively promote the generating and developing processes of sperms of testicular seminiferous epithelium and increase the sperm counts, which has the effects of tonifying the spleen and kidney, invigorating the circulation of blood and promoting the spermatogenic process.