| Objective: To observe the effect of Guiyao Shengjing Capsule on the expression of Bcl-2,Bax,Caspase-9 and Caspase-3 in the testis of rats with Cyclophosphamide(CTX)-induced infertility(Male Infertility,MI).Method: 50 SPF male SD rats were randomly selected as the blank group and 40 as the model group.The blank group was intraperitoneally injected with normal saline,and the model group was intraperitoneally injected with CTX(40mg/kg)for 5 consecutive days to establish MI Rat model.After modeling,3 rats in the blank group and 5 rats in the model group were randomly selected for model verification.After confirming that the modeling was successful,the rats in the model group were divided into model group,positive control group,low-dose guiyao group,middle-dose guiyao group and high-dose guiyao group,and the intervention was started.The blank group and the model group were given normal saline(1m L/d)by gavage,the positive control group was given valsartan capsules(1.44mg/m L/d)by gavage,and the guiyao group was given low,medium and high doses of Gui medicine Shengjing capsules(28.5mg/ml/d,57mg/ml/d,114mg/ml/d)gavage for 4 consecutive weeks.After the intervention,the body weight of the rats was measured.The testes and epididymis were removed under aseptic conditions.The sperm parameters of the rat epididymis were measured by the diffusion method,the changes in the tissue structure of the rat testis were observed by the HE staining method,and the IHC staining method was used for detection.The expression levels of Bcl-2,Bax,Caspase-9 and Caspase-3 in testis tissues of rats in each group.Result:1.Model verification: The body mass,testis and epididymis mass,sperm density,motility sperm percentage,and forward motility sperm percentage of the model group were significantly lower than those of the blank group.The testicular tissue structure of the model group showed obvious pathological changes.The preparation of the mouse model meets the MI model standard.2.Pathological observation of rat testis: the testicular tissue structure of the low,medium and high dose guiyao group was significantly improved compared with the model group and the positive control group,the seminiferous tubules tended to be regular,the basement membrane thickened,fibromuscular cells and supporting cells The number is evenly distributed,the arrangement of spermatogonia and spermatocytes tends to be neat,the number of layers increases,the damage of the outer cell membrane can be repaired,the number of sperm cells and sperm increases,the blood supply of capillaries is improved,and the infiltration of inflammatory cells is reduced.Among them,there was no obvious pathological change in the morphological structure of testis in the high-dose guiyao group,and the improvement was the most obvious,and there was no significant difference compared with the blank group.3.Rat body mass: After intervention,the body mass of the guiyao group and the positive control group was significantly higher than that of the model group(P<0.01),and the middle and high-dose guiyao group was better than the positive control group(P<0.05).There was no significant difference in body weight between the high-dose guiyao group and the blank group(P>0.05).4.The quality of rat testis and epididymis:(1)The quality of testis:the quality of testis in the guiyao group and the positive control group was significantly higher than that of the model group(P<0.01),and the high-dose guiyao group was better than the positive control group(P<0.05)There was no significant difference in testicular quality between the high-dose guiyao group and the blank group(P>0.05).(2)The quality of epididymis: the quality of epididymis in the guiyao group and positive control group was increased compared with the model group(P<0.05),and the high-dose guiyao group was better than the positive control group(P<0.05).There was no significant difference in quality of epididymis between the high-dose guiyao group and the blank group(P>0.05).5.Rat sperm parameters:(1)Sperm density: The sperm density of the guiyao group and the positive control group was significantly higher than that of the model group(P<0.01),and the guiyao group was better than the positive control group(P < 0.01).The sperm density of the high-dose guiyao group was compared with the blank group.There was no significant difference in comparison(P>0.05).(2)Percentage of motile sperm: the percentage of motile sperm in the guiyao group and the positive control group was significantly higher than that of the model group(P<0.01),and the middle and high dose guiyao group was better than the positive control group(P<0.01),and the high dose of guiyao There was no significant difference in the percentage of motile sperm in the group and the blank group(P>0.05).(3)The percentage of forward motile sperm: The percentage of forward motile sperm in the guiyao group and the positive control group was significantly higher than that of the model group(P<0.01),and the middle and high-dose guiyao group was better than the positive control group(P<0.01).There was no significant difference in the percentage of forward motile sperm in the middle and high-dose groups of guiyao compared with the blank group(P>0.05).6.The expression of Bcl-2 in rat testis: the positive expression rate of Bcl-2 in the guiyao group and the positive control group was significantly higher than that of the model group(P<0.01),and the guiyao group was better than the positive control group(P< 0.05),there was no significant difference in the positive expression rate of Bcl-2between the high-dose guiyao group and the blank group(P>0.05).7.Bax expression in rat testis: the positive expression rate of Bax in the guiyao group and the positive control group was significantly lower than that of the model group(P<0.01),and the middle and high-dose guiyao group was better than the positive control group(P<0.01).8.The expression of Caspase-9 in rat testis: the positive expression rate of Caspase-9 in the guiyao group and the positive control group was significantly lower than that of the model group(P < 0.01),and the middle and high-dose guiyao group was better than the positive control Group(P<0.01).9.The expression of Caspase-3 in rat testis tissue: The positive expression rate of Caspase-3 in the guiyao group and the positive control group was significantly lower than that of the model group(P<0.01),and the high-dose guiyao group was better than the positive control group(P<0.05).Conclusion:Guiyao Shengjing Capsules can improve the pathological damage of the testis of MI rats,increase the body mass,testis and epididymis mass,total sperm count and motility of MI rats,and can up-regulate the expression of Bcl-2 in testicular tissues,and down-regulate Bax,Caspase-9,and Caspase-9.The expression of Caspase-3 has a significant dose-effect relationship,which may be one of the mechanisms of Guiyao Shengjing Capsules in treating MI. |
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