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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线粒体功能的影响。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体质量给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6天开始按体质量分别给SG和LG小鼠灌服益精方,剂量分别为人类常规用量(以60 kg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34 d,CG常规饲养,然后对小鼠进行精液常规分析,流式细胞术检测精子线粒体功能。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/m L;精子活力[(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)%;精子活率[(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、精子活力、精子活率显著低于CG组(P<0.05);经治疗后LG组精子密度、精子活力和精子活率明显增加,与MG组比较有显著性差异(P<0.05),而与CG组在精子活力和精子活率上没有明显区别。在CG、MG、SG和LG组,线粒体功能良好精子的百分率分别为(1105.93±0.33)%、(848.55±6.15)%、(888.77±22.46)%、(1000.885±72)%,线粒体功能丧失精子的百分率分别为(27.94±0.32)%、(42.43±1.00)%、(35.11±1.23)%、(30.91±0.44)%,MG小鼠精子线粒体功能良好的表达量明显下降,而线粒体功能丧失的表达量明显增多,与CG小鼠比较均有明显差异(P<0.05);经治疗后,LG小鼠精子线粒体功能良好的表达量显著性增加(P<0.05),同时线粒体功能丧失的表达量明显减少,与MG小鼠比较有明显差异(P<0.05)。结论腹腔注射环磷酰胺可使小鼠精子密度、精子活力、精子活率降低,大剂量益精方可以增加小鼠精子密度,并增加精子线粒体功能,提高精子活力和精子活率,从而达到治疗少弱精子症的目的。
Objective To observe the effects of Yijing Recipe on mitochondrial function of spermatozoa in mice with hypoweak spermatology induced by cyclophosphamide. Methods Forty male Kunming mice were randomly divided into control group (CG), model group (MG), low dose Chinese medicine treatment group (SG) and large dose Chinese medicine treatment group (LG). Intraperitoneal injection of physiological saline (NS) was given to CG mice at 60 mg/kg body weight. MG, SG, and LG mice were injected intraperitoneally with cyclophosphamide once a day for 5 days. On the 6th day, SG was administered according to body weight. LG mice were dosed with Yijing decoction at doses of 1 and 5 times the normal human dose (60 kg as the standard). MG mice were given equal volume of NS gavage once daily for 34 days. CG routine After feeding, the mice were routinely analyzed for semen and flow cytometry was used to detect sperm mitochondrial function. Results The epididymal sperm density in CG, MG, SG and LG mice was (5.20±1.34), (1.73±0.03), (2.08±0.01), (3.31±0.56)×106/m L, respectively; sperm motility [(a) Percentage of +b) spermatozoa were (14.49±0.30)%, (6.64±1.88)%, (11.99±1.01)%, (19.40±3.13)%, respectively; spermatozoa survival rate [(a+b+c) grade spermatozoa Percentages were (68.39±15.13)%, (39.96±4.89)%, (62.28±4.43)%, (73.61±5.05)%, respectively. The sperm density, sperm motility and sperm motility in MG group were significantly lower than those in CG group (P<0.05). After treatment, the sperm density, sperm motility and sperm motility in LG group increased significantly, which was significantly different from that in MG group (P< 0.05), but there was no significant difference in sperm motility and sperm viability with the CG group. In the CG, MG, SG, and LG groups, the percentage of spermatozoa with good mitochondrial function was (1105.93±0.33)%, (848.55±6.15)%, (888.77±22.46)%, (1000.885±72)%, respectively, and mitochondrial loss of spermatozoa. The percentage of mitochondrial function was significantly decreased in MG mice, but the percentage of mitochondrial function was significantly decreased (27.94±0.32)%, (42.43±1.00)%, (35.11±1.23)%, (30.91±0.44)%. The expression level was significantly increased compared with CG mice (P<0.05). After treatment, the expression of sperm mitochondrial function in LG mice was significantly increased (P<0.05), and the expression of mitochondrial function loss was also observed. Significantly decreased, compared with MG mice there was a significant difference (P <0.05). Conclusion Intraperitoneal injection of cyclophosphamide can reduce sperm density, sperm motility and sperm viability in mice. High-dose Yijing Recipe can increase sperm density in mice, increase sperm mitochondrial function, increase sperm motility and sperm viability, and thus achieve therapeutic effects. The purpose of less severe sperm disease.