| Objective1.To observe the clinical efficacy and safety of Erxianzanyu prescription in the trea tment of asthenospermia,and provide evidence-based medical evidence of imPRoving the clinical efficacy of asthenospermia by tonifying kidney,strengthening spleen,removing bloo d stasis and removing turbidity.2.To explore the mechanism of Erxianzanyu prescription in the treatment of astheno spermia,namely,whether Erxianzanyu prescription adjusts the Bcl-2/Bax-Caspase9-Caspase3 pathway,reduces the sperm cell apoptosis caused by this pathway,and then treats asthenos permia,pfoviding experimental data basis for the treatment of asthenospermia by tonifying the kidney,strengthening the spleen,removing blood stasis and removing turbidity.Method1.Clinical trial studies:72 patients with asthenospermia who met the inclusion criteri a were randomly divided into control group and treatment group,with 36 patients in each group.The control group was treated with 1-carnitine oral liquid for 3 months,and the tre atment group was treated with Erxianzanyu prescription for 3 months.Clinical data were collected by detecting sperm density,PR grade sperm,PR+NP grade sperm,semen volume,T CM syndrome score,pregnancy rate,safety index and other indicators.2.Animal experiment studies:24 male SD rats were divided into 6 groups by rando m sampling:blank group,model group,low-dose group,medium-dose group,high-dose group and 1-carnitine control group,with 4 rats in each group.To observe the effects of Erxianza nyu prescription on sperm quality,epididymis electron microscope structure,mitochondrial a ctivity,gene transcription and protein expression in asthenospermia model rats.Result1.Clinical research:A total of 72 subjects were included in this study,including 36 in the control group and 36 in the treatment group.During the study period,there were 8 cases of voluntary w ithdrawal in the control group and 2 cases of shedding in the treatment group due to in complete foIlow-up.Finally,there were 28 cases in the control group and 34 cases in the t reatment group,a total of 62 cases were included in the statistics.No adverse reactions we re reported in all cases,and the specific results were as follows:1.1 Evaluation of main efficacy indicators1.1.1 Sperm motilityForward motile sperm(PR)and total sperm motility(PR+NP)in both groups were s ignificantly increased after treatment(P<0.01).Compared with the control group,the ther apeutic effect of the treatment group was better(P<0.05).1.2 Evaluation of secondary efficacy indicators1.2.1 Semen volumeAfter treatment,semen volume was improved in both groups(P<0.05),and the impr ovement in treatment group was better than that in control group(P<0.05).1.2.2 Sperm concentrationThe sperm concentration in the control group changed significantly after treatment(P<0.05),and that in the treatment group changed significantly after treatment(P<0.01).After treatment,the improvement of sperm concentration in treatment group was better th an that in control group(P<0.05).1.2.3 TCM related index scoringThe total score of TCM syndromes in the control group had no statistical significan ce before and after treatment(P>0.05).The total score of TCM syndrome in the treatme nt group was significantly improved after treatment(P<0.01).In the treatment group,there were significant therapeutic effects on eight kinds of s ymptoms(P<0.01),such as soft waist and knee,fear of cold and cold limbs,listlessness,severe white face,apathy of sexual desire,forgetfulness,pain in perineum/lower abdome n and loose stools.The treatment group had obvious effect in the treatment of two kinds of symptoms,such as clear urine and loss of appetite(P<0.05);In the control group,th ere was no statistical significance in these symptom scores after treatment compared with before treatment(P>0.05).1.3 Clinical efficacy resultsThe total effective rate was 71.4%in the control group and 88.2%in the treatment group,PRoving that the therapeutic effect of the treatment group was significantly better t han that of the control group(P<0.01).1.4 Safety ObservationThe subjects in the control group and the treatment group showed no adverse reacti ons during the test period,and no significant abnormalities in blood routine,urine routine,li ver and kidney function and renal function were observed after the study.2.Animal experimental studies2.1 Histology of rat testis epididymis under electron microscopeBlank group:no mitochondrial swelling;Model group:mitochondrial edema and frac ture;Low dose Erxianzanyu prescription group:mitochondria and endoplasmic reticulum swelling;The mitochondria of Erxianzanyu prescription medium group were normal.Erxi anzanyu prescription high-dose group:no swelling of mitochondria and endoplasmic retic ulum;In the 1-carnitine group,mitochondria were normal.2.2 Sperm count and motility of ratsCompared with blank group,the number of viable sperm in model group was signifi cantly decreased(P<0.05),indicating that asthenospermia rats were successfully modeled.Compared with model group,the sperm concentration of Erxianzanyu prescription low-do se,medium-dose,high-dose and 1-carnitine groups was significantly improved(P<0.05);C ompared with model group,the sperm survival rate of Erxianzanyu prescription low-dose,medium-dose,high-dose and 1-carnitine groups was significantly increased(P<0.05).2.3 Effects of ROS level in rat testicular tissueROS level of testicular tissue was assessed by DHE staining.Ornidazole could aggra vate oxidative damage and enhance fluorescence expression in testicular tissue cells,whil e Ercienzanyu prescription and 1-carnitine could reduce fluorescence expression in testicul ar tissue cells of rats.2.4 expression of Bcl-2,Bax,Caspase3 and Caspase9 mRNA in rat testicular tissueCompared with blank group,the expression of Bcl-2 mRNA in spermatogenic cells of model group was increased(P<0.05);Compared with model group,Erxianzanyu prescr iption low-dose,medium-dose,high-dose and 1-carnitine groups decreased(P<0.05).Compared with blank group,the mRNA expression levels of Bax,Caspase3 and Cas pase9 in spermatogenic cells of model group were decreased(P<0.05);Compared with m odel group,Erxianzanyu prescription low-dose group,medium-dose group,high-dose grou p and 1-carnitine group increased significantly(P<0.05).2.5 protein expressions of Bcl-2,Bax,CYTC,Caspase3 and Caspase9 in rat testicular tissueCompared with blank group,the expression level of Bcl-2 in model group was incre ased(P<0.05);The expression levels of Bax,CYTC,Caspase3 and Caspase9 decreased(P<0.05).Compared with model group,the expression level of Bcl-2 in Erxianzanyu prescripti on group and 1-carnitine group was decreased(P<0.05);The expression levels of Bax,C YTC,Caspase3 and Caspase9 were increased(P<0.05).Conclusion1.The clinical research verified that Erxianzanyu prescription has obvious efficacy a nd safety in imPRoving semen volume,sperm concentration and sperm motility through c omparative observation of different treatment methods for enrolled cases:1.1 Erxianzanyu prescription can effectively increase sperm motility,improve sperm d ensity and increase semen volume.1.2 Erxianzanyu prescription can effectively reduce the TCM syndrome score of pati ents with asthenospermia and improve the clinical comprehensive symptoms of patients.1.3 Erxianzanyu prescription has high safety in treating patients with asthenospermia.2.In Experimental study through ORN intragastric modeling,mature technology,can c ause epididymis damage,resulting in decreased sperm motility in rats.2.1 Erxianzanyu prescription can improve epididymal damage of orn-induced astheno spermia in rats and improve the sperm motility of rats,and the effectiveness of Erxianzan yu prescription in treating asthenospermia was preliminarily verified.2.2 Erxianzanyu prescription can reduce oxidative stress damage of sperm in asoospe rm infertility rats,down-regulate bcl-2 mRNA and protein expression,and up-regulate Bax,Caspase3,Caspase9 mRNA and protein expression,suggesting that Erxianzanyu prescription can induce changes of Bax and Bcl-2 genes.Activation of Caspase 9-Caspase 3 pathway improves semen quality. |
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