| Objective: To observe the therapeutic effect of L-carnitine,compound Xuanju capsule and their combination on oligospermia induced by cyclophosphamide in rats and its related mechanism.The aim of this study is to provide a theoretical basis for the treatment of clinical male infertility.Methods:A total of 40 SPF male Wistar rats aged 8 weeks,weighing(232±23g),were randomly divided into 5 groups,8 rats in each group,respectively: Group A(blank group),group B(cyclophosphamide group),group C(cyclophosphamide +L-carnitine group),group D(cyclophosphamide +compound Xuanju capsule group),group E(cyclophosphamide + L-carnitine + compound Xuanju capsule group).Groups B,C,D and E were intraperitoneally injected with 40mg/kg cyclophosphamide once A day,and group A was intraperitoneally injected with the same amount of normal saline for 5 days.At the end of the modeling stage,the intervention of each group began the next day,that is,groups A and B were intraperitoneally injected with normal saline and gavage,group C was intraperitoneally injected with 100mg/kg L-carnitine and gavage of normal saline.Group D was intraperitoneally injected with normal saline and 40mg/kg compound Xuanju capsule solution by gavage,and group E was intraperitoneally injected with 100mg/kg L-carnitine and 40mg/kg compound Xuanju capsule solution by gavage,once a day for 28 days.Sperm in the epididymis were collected for analysis of sperm quality and motility using a blood cell counting plate.One testis of the rats was stained with HE to observe the histopathological changes,and the other testis was examined for oxidative stress related indicators: The contents of reactive oxygen species(ROS),nitric oxide(NO),catalase(CAT),superoxide dismutase(SOD),reduced glutathione(GSH),malondialdehyde(MDA)and the expression of estrogen receptor α(ERα)protein in testicular tissue were evaluated.The therapeutic effects of each intervention program were evaluated by statistical comparison of experimental data in each group.Results:1.Body weight,testis and epididymis weight of rats in each group: L-carnitine,compound Xuanju capsule and L-carnitine combined with compound Xuanju capsule intervention all alleviated cyclophosphamide-induced reduction in body weight,testis and epididymis weight(P < 0.01).The efficacy of L-carnitine combined with compound Xuanju capsule in alleviating weight loss was significantly better than that of L-carnitine and compound Xuanju capsule monotherapy(P < 0.05).Compared with the two monotherapy,the efficacy of L-carnitine in alleviating weight loss was significantly better(P < 0.01).There was no significant difference in the efficacy of three intervention programs in alleviating the reduction of the weight of the testis and epididymis(P > 0.05).2.Comparison of semen quality in each group: The combined intervention of L-carnitine,compound Xuanju capsule and L-carnitine + Compound Xuanju capsule could alleviate the decrease of sperm count and motility caused by cyclophosphamide(P < 0.01).The combined intervention of L-carnitine + compound Xuanju Capsule had significant advantages in alleviating sperm count and sperm motility(P < 0.05),while there was no significant difference between the two monotherapy treatments in alleviating sperm count and sperm motility(P > 0.05).3.Histomorphological comparison of testis in each group: L-carnitine,compound Xuanju capsule and L-carnitine combined with compound Xuanju capsule could alleviate cyclophosphamide-induced testicular pathological damage(P < 0.05).The combination of L-carnitine + compound Xuanju capsule had a significant advantage in reducing the pathological damage of testicular tissue compared with the two monotherapy(P < 0.05),and there was no significant difference in the efficacy of the two monotherapy in reducing the pathological damage of testicular tissue(P > 0.05).4.Detection of oxidative stress markers in testicular tissue of rats in each group:(1)Contents of ROS,NO and MDA: L-carnitine,compound Xuanju capsule and L-carnitine combined with compound Xuanju capsule all alleviated cyclophosphamide-induced increases in the contents of ROS,NO and MDA in the testicular tissue(P < 0.05).L-carnitine combined with compound Xuanju capsule had a significant advantage in reducing the contents of ROS,NO and MDA in the testicular tissue(P < 0.05).Compared with the two single drug therapies,L-carnitine had a significant advantage in reducing the contents of ROS and NO in the testicular tissue(P < 0.01).There was no significant difference in the efficacy of the two monotherapy in reducing the content of MDA in testicular tissue(P > 0.05).(2)CAT and SOD activities: L-carnitine,compound Xuanju capsule and L-carnitine combined with compound Xuanju capsule all alleviated cyclophosphamide-induced reduction of CAT and SOD activities in testicular tissue(P < 0.05).The combination of L-carnitine and compound Xuanju capsule had a significant advantage in improving the activities of CAT and SOD in the testicular tissue compared with the two monotherapy(P <0.05),and there was no significant difference between the two monotherapy in improving the activities of CAT and SOD in the testicular tissue(P > 0.05).(3)GSH content: Compound Xuanju capsule and L-carnitine combined with compound Xuanju capsule could alleviate cyclophosphamide-induced reduction in testicular GSH content(P < 0.01),and there was no significant difference in the efficacy of L-carnitine combined with compound Xuanju capsule in improving testicular GSH content compared with compound Xuanju capsule(P > 0.05).5.The expression of ERα protein in the testis tissue of each group: Compound Xuanju capsule and L-carnitine combined with compound Xuanju capsule could increase the expression level of ERα protein(P < 0.01),and the combined intervention of L-carnitine and compound Xuanju capsule had a significant superiority over compound Xuanju capsule in improving the expression level of ERα protein in the testis tissue(P < 0.01).Conclusion:1.The combined treatment of L-carnitine,compound Xuanju capsule and L-carnitine + compound Xuanju capsule has a significant repair effect on spermatogenic function in rats with oligozoospermia induced by cyclophosphamide,and the combined treatment of L-carnitine and compound Xuanju capsule is better than either L-carnitine or compound Xuanju capsule alone;2.The possible mechanism of L-carnitine combined with compound Xuanju capsule in the treatment of oligozoospermia induced by cyclophosphamide is to reduce the levels of ROS,NO and MDA in the testicular tissue,and increase the activities of CAT and SOD,GSH content and the expression of ERα protein in the testicular tissue. |
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