“益精方”对小鼠精子尾部特异性钙通道CatSper1的影响

Yijingfang increases the expression of CatSper1 in mice

目的:观察"益精方"对环磷酰胺所致少弱精子症模型小鼠精子尾部特异性钙通道CatSper1的影响。方法:将40只雄性昆明小鼠随机分为对照组(CG)、模型组(MG)、小剂量中药治疗组(SG)和大剂量中药治疗组(LG),按60mg/kg体重给CG小鼠腹腔注射生理盐水(NS),给MG、SG、LG小鼠腹腔注射环磷酰胺,每天1次,连续5d,第6d开始按体重分别给SG和LG小鼠灌服"益精方",剂量分别为人类常规用量(以60kg为标准)的1倍和5倍,MG小鼠给予等体积的NS灌胃,每天1次,连续34d,CG常规饲养,然后对小鼠进行精液常规分析,并用RT-PCR法检测小鼠精子CatSper1的表达。结果:CG、MG、SG和LG组小鼠附睾精子密度分别为(5.20±1.34)、(1.73±0.03)、(2.08±0.01)、(3.31±0.56)×106/ml,a+b级精子百分率分别为(14.49±0.30)%、(6.64±1.88)%、(11.99±1.01)%、(19.40±3.13)%,a+b+c级精子百分率分别为(68.39±15.13)%、(39.96±4.89)%、(62.28±4.43)%、(73.61±5.05)%,MG组精子密度、a+b级精子百分率、a+b+c级精子百分率显著低于CG组(P<0.05),经治疗后LG组精子密度、a+b级精子百分率、a+b+c级精子百分率明显增加,与MG组比较有显著性差异(P<0.05),而与CG组在a+b级精子百分率和a+b+c级精子百分率上没有明显区别。CatSper1的表达量在CG、MG、SG和LG组分别为0.76±0.05、0.73±0.03、0.75±0.12、0.85±0.04,LG组显著高于MG组(P<0.05),而与CG组比较没有明显区别。结论:腹腔注射环磷酰胺可使小鼠精子密度、a+b级精子百分率、a+b+c级精子百分率降低,CatSper1表达量下降,大剂量"益精方"可以通过提高CatSper1的表达量,增加小鼠精子密度、a+b级和a+b+c级精子百分率,从而达到治疗少弱精子症的目的。

Objective：To observe the effects of Yijingfang on CatSper1 in the mouse model of cyclophosphamide-induced oligoasthenospermia.Methods：Forty Kunming male mice were randomly divided into a control group（CG）,a model group（MG）,a small-dose Yijingfang group（SG）,and a large-dose Yijingfang group（LG）.The mice of CG were intraperitoneally injected with normal saline at 60 mg/kg once a day,while those of MG,SG and LG with cyclophosphamide,all for 5 days.During the next 34 days,the mice of SG and LG received intragastric administration of Yijingfang once a day,the former at a dose 2 times and the latter 5 times that of human routine usage,those of MG given the same volume of normal saline,and CG normally fed.At 35 days,we measured the sperm count,percentages of grades a＋b and a＋b＋c sperm,and the expression of CatSper1 in the epididymal sperm of the mice.Results：The sperm counts of CG,MG,SG and LG were（5.20±1.34）,（1.73±0.03）,（2.08±0.01）and（3.31±0.56）×106/ml,respectively,significantly lower in MG than in CG（P0.05）,but higher in LG than in MG（P0.05）.The grade a＋b sperm constituted（14.49±0.30）,（6.64±1.88）,（11.99±1.01）and（19.40±3.13）% in CG,MG,SG and LG,respectively,remarkably lower in MG than in CG（P0.05）but higher in LG than in MG（P0.05）;the grade a＋b＋c sperm accounted for（68.39±15.13）,（39.96±4.89）,（62.28±4.43）and（73.61±5.05）%,respectively,obviously lower in MG than in CG（P0.05）but higher in LG than in MG（P0.05）;the CatSper1 expressions were 0.76±0.05,0.73±0.03,0.75±0.12 and 0.85±0.04,respectively,markedly higher in LG than in MG（P0.05）.Conclusion：Intraperitoneal injection of cyclophosphamide decreases the sperm count,percentages of grades a＋b and a＋b＋c sperm,and the expression of CatSper1 in mice,while large-dose Yijingfang can increase the above parameters,and hence contributes to the treatment of oligoasthenospermia.