益肾活血方对精子活力低下症病理模型大鼠血清睾丸活性酶的影响

Effects of Yishen Huoxue Decoction on Serum Testicular Enzyme Activity in Rats with Hypomotility of Sperm

目的观察益肾活血方对精子活力低下症病理模型大鼠精子活动力及血清睾丸活性酶的影响,初步阐述益肾活血方改善精子活动力的部分作用机制。方法将60只雄性Wistar大鼠随机分成正常组和模型组以及中药低、中、高剂量组,每组大鼠各12只。除正常组以外,模型组、中药各剂量组的大鼠予灌服西药雷公藤多苷片5周以造模。经成功建立精子活动低下症大鼠病理模型后,中药高、中、低各剂量组分别予益肾活血方,按5、10、15 g/kg/天调配,并进行中药灌胃5周,正常组、模型组均予普通饲养。末次给药后立即取各组大鼠的附睾尾进行精子质量相关指标检测,并采用酶联免疫法(ELISA)检测大鼠血清中顶体酶(ACE)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、乳酸脱氢酶(LDH)、透明质酸酶(HYD)、γ-谷氨酸转肽酶(γ-GT)的浓度变化情况。结果正常组、低剂量组、中剂量组、高剂量组模型大鼠的精子总活率明显高于模型组(P<0.05);高剂量组模型大鼠的a级精子活力明显高于模型组(P<0.05);低剂量组、中剂量组、高剂量组模型大鼠的b级精子活力、a级+b级精子活力明显高于模型组(P<0.05)。正常组、低剂量组、中剂量组的ACE和AKP及LDH浓度均明显高于模型组(P<0.05),高剂量组的ACE浓度明显高于模型组(P<0.05),高剂量组的AKP和LDH浓度与模型组相比,均无显著性差异(P>0.05);正常组及治疗各剂量组的ACP浓度明显低于模型组(P<0.05);正常组以及中、高剂量组的HYD浓度高于模型组(P<0.05),低剂量组的HYD浓度与模型组相比无明显差异(P>0.05);正常组及治疗各剂量组的γ-GT浓度与模型组相比,无显著性差异(P<0.05)。结论益肾活血方显著提升实验大鼠精子的活动力,其改善病理模型大鼠精子活动力的调控机制可能是通过提升血清ACE、HYD、AKP及LDH的浓度,从而提高精子活力低下症病理模型大鼠精子的活动力。

Objective To observe the effects of Yishen Huoxue Decoction on sperm motility and serum testicular active enzymes in pathological model rats with hyposperm motility,and to expatiate the partial mechanism of Yishen Huoxue Decoction in improving sperm motility.Methods Sixty male Wistar rats were randomly divided into normal group,model group,low-dose,medium-dose and high-dose TCM groups,with 12 rats in each group.Except for the normal group,the rats in the model group and the traditional Chinese medicine groups were given western medicine Tripterygium wilfordii polyglycosides tablet for 5 weeks to establish the model.After the successful establishment of the pathological model of hyposperm motility,the high-dose,medium-dose and low-dose groups of Chinese medicine were given Yishen Huoxue Decoction(5,10,15 g/(kg·d))respectively,and given Chinese medicine by intragastric administration for 5 weeks.The normal group and model group were fed by ordinary feeding.Immediately after the last administration,caudal epididymis of rats in each group was collected for sperm quality related indexes detection.Serum concentrations of acrosomal enzyme(ACE),acid phosphatase(ACP),alkaline phosphatase(AKP),lactate dehydrogenase(LDH),hyaluronidase(HYD)andγ-glutamate transpeptidase(γ-GT)were determined by enzyme-linked immunosorbent assay(ELISA).Results The total sperm motility in normal group,low dose group,medium dose group and high dose group was significantly higher than that in model group(P<0.05).The A sperm motility of model rats in high-dose group was significantly higher than that in model group(P<0.05).The B class sperm motility and A+B class sperm motility in lowdose,medium-dose and high-dose groups were significantly higher than those in model group(P<0.05).The concentrations of ACE,AKP and LDH in normal group,low dose group and medium dose group were significantly higher than those in model group(P<0.05).The concentrations of ACE in high dose group were significantly higher than those in model group(P<0.05).The concentrations of AKP and LDH in high dose group were not significantly different from those in model group(P>0.05).ACP concentration in normal group and treatment dose groups was significantly lower than that in model group(P<0.05).The concentration of HYD in normal group,medium-dose and high-dose groups was higher than that in model group(P<0.05),and there was no significant difference between the concentration of HYD in low-dose group and model group(P>0.05).Compared with the model group,there was no significant difference inγ-GT concentration between the normal group and the treatment dose groups(P<0.05).Conclusion Yishen Huoxue Decoction significantly improves the motility of sperm in experimental rats,and the regulation mechanism of improving the motility of sperm in pathological model rats may be through increasing the concentrations of ACE,HYD,AKP and LDH in serum,thus improving the motility of sperm in pathological model rats with hyposperm.