加味聚精食疗方对DS动物模型精子质量及睾丸组织中CR16表达的影响

Effect of Jiawei Jujing Shiliao Prescription on Quality of Sperms and CR16 Expression in Oligozoospermia or Asthenospermia Rats

目的：观察加味聚精食疗方对少、弱精子症小鼠睾丸组织中CR16表达的影响。方法：60只8周龄雄性小鼠适应性饲养1周后随机编号,采取区组随机化法将小鼠分为6组：正常组（G1）、DS模型空白对照组（G2）、低剂量治疗组（G3）、中剂量治疗组（G4）、高剂量治疗组（G5）、预防治疗组（G6）,每组10只。G2~G6组通过尾静脉单剂量注射5-FU（100 mg/kg）,14 d后形成稳定的DS模型。G1正常组全程给予蒸馏水4 g/kg灌胃,连续40 d;G2空白对照组及G3~G5治疗组自造模开始即给予蒸馏水4 g/kg灌胃,G6预防治疗组给予加味聚精食疗方6.3 g/kg灌胃,连续14 d。G2~G6组造模完成后,G2组予蒸馏水4 g/kg灌胃;G3~G5组分别予加味聚精食疗方低剂量（3.15 g/kg）灌胃、中剂量（6.3 g/kg）灌胃、高剂量（12.6 g/kg）灌胃;G6组继续蒸馏水4 g/kg连续灌胃。以上均1次/d,连续喂养26 d。各组治疗结束后作集中处死,计算机辅助精子分析（CASA）技术检测小鼠附睾精子质量;对小鼠肝、肾器官及睾丸组织进行病理学检查;免疫组织化学S-P法检测基因CR16在SD动物模型体内的表达。结果：在精子质量的改善方面,G2、G6组比较无明显差异,不具有统计学意义（P〉0.05）。G3、G4、G5组均较G2、G6组有明显改善（P〈0.05）,且G4组优于G3、G5组（P〈0.05）。在阴性对照无特异性着色的情况下对结果进行判读,G3、G4、G5组小鼠睾丸组织生精小管上皮中CR16均有不同程度的表达,明显高于G2、G6组,差异具有统计学意义（P〉0.05）,G4组小鼠睾丸组织结构修复情况、睾丸组织CR16的表达水平（染色程度）均明显上调。镜下观察见小鼠睾丸组织中CR16阳性反应呈棕褐色或浅棕色颗粒状,散在分布于支持细胞胞浆胞膜。结论：加味聚精食疗方能够一定程度上提高附睾精子的质量,安全无不良反应,其通过上调DS动物模型体内CR16的表达水平可能是治疗少、弱精子症的作用机制之一。

Objective： To observe the influence of Jiawei Jujing Shiliao Prescription on the CR16 expression in oligozoospermia spermatozoa rats. Methods： Sixty rats were randomized into six groups,with ten rats in each group. Among them,rats in Group 2 to Group 6 were administered with 5- FU iv（ 100 mg / kg） for fourteen days to establish the DS models. Rats in Group 1 were given distilled water（ 4 g / kg） for continuous 40 days. After model establishment,rats in Group 2 and Group 6 were intragastrically administered with distilled water（ 4 g / kg） and rats in Group 3 to Group 5 were intragastrically administered with Jujing Shiliao Prescription with low dose（ 3. 15 g / kg）,moderate dose（ 6. 3 g / kg） and high dose（ 12. 6 g / kg） respectively,for 26 days,and in Group 6,before model establishment,Jujing Shiliao Prescription（ 6. 3 g/kg） was applied for 14 days. All the rats were sacrificed after treatments. The sperm quality in the epididymis was detected with computer- assisted sperm analysis（ CASA） technique and the pathological changes of the liver,kidney and testes were observed. The CR16 expression in the rats was explored with immunohistochemical method. Result： Forthe sperm quality,there was no significant difference between Group 2 and Group 6（ P 〉0. 05）. Compared with those in Group 2 and Group 6,there was significant improvement in Group 3,4 and 5（ P〈 0. 05）,with Group 4 better than Group3 and Group 5（ P 〈0. 05）. The CR 16 expression in Group 3,4 and 5 was apparently higher than those in Group 2 and 6（ P 〈0. 05）. Conclusions： Jiawei Jujing Shiliao Prescription could improve the sperm quality in the epididymis. Meanwhile,it could up- regulate the expression of CR16,which may be one of the mechanisms for the treatments of oligozoospermia spermatozoa.