Clinical And Experimental Study Of Guangyu Yulin Decoction In Treating Kidney-Accent Deficiency And Weak Sperm Disease

OBJECTIVEMale infertility has become a global medical and sociological problem that needs safe and effective treatments among which the most common form is asthenospermia.Western medicine treatment is not ideal,while Chinese medicine has unique advantages.Guangsiyulin Decoction is created from Wuziyanzong pills by the addition of some other drug which is used in clinic for 20 years and shows a certain effect,while there is no relevant research about it.Therefore,this study intends to evaluate the efficacy of Guangsiyulin Decoction in the treatment of patients with male infertility of oligozoospermia diagnosed as renal insufficiency in TCM.Then we design the animal experiment to observe impact of the function on the spermatogenesis disorder model and elaborate its mechanism of action by using Guangsiyulin Decoction.METHODClinical study:A clinical study was performed to observe the semen quality(Semen volume,liquefaction,sperm concentration,total sperm count,total sperm viability,forward percentage of motile spermatozoa)and the influence of TCM syndrome scores.The total duration of treatment of Guangsiyulin Decoction was 90 days performed daily.The observed points were treated in 0,30,60,and 90 day.The patients’ TCM syndrome scores and semen quality were recorded in each observed points.During the clinical study period,adverse events were strictly recorded.In each observed points,we would assess and deal with the shedding and removal.Finally,we would count all data results.Experimental study:1.The rat model of spermatogenesis was induced by Tripterygium glycoside;2.The rats were randomly divided into blank control group,model control group,and traditional Chinese medicine intervention group with 12 rats in each group.Guangsiyulin Decoction granule suspension(10ml/kg)was intragastrically administered in traditional Chinese medicine intervention group.The blank control group and the model control group were administered with deionized water.All administration was lasted 4 weeks.3.Observed indicators:general condition,body weight,organ coefficient,semen quality,and serum sex hormone levels in the rat model of spermatogenesis disorder,testicular tissue pathology,testicular tissue and ultrastructure of spermatogenic cells using Guangsiyulin Decoction.The expression of Bax and Bcl-2 protein in rat testis was detected by immunohistochemistry.The protein and mRNA expression of c-kit and Oct4 were detected by western blot and Real-Time PCR.RESULTClinical studies:After 90 days,intervention in 50 patients,the semen quality including liquefaction status,total live rate(PR+NP),sperm concentration,total number and other evaluation indicators were improved substantially that had statistically significant(P<0.05 or P<0.01);The syndrome scores and total scores of various syndromes of kidney essence deficit deficiency were significantly decreased as the progress of treatment going on(P<0.05 or P<0.01).There were no case of shedding,rejection and adverse events in the entire observation period did notExperimental study:Comparing with the model control group,the general state of the rat,the coefficient of the viscera of the epididymis,the index of the semen quality,follicle-stimulating hormone(FSH),luteinizing hormone(LH),estosterone(T)and Estradiol(E2)were improved in the group of Guangsiyulin Decoction significantly(P<0.05 or P<0.01);Compared with normal group in the pathological morphology and ultrastructure of the testis,the structure and arrangement of seminiferous tubules were more normal,and there were more obvious spermatogenic cells in the spermatogenesis;degeneration and apoptotic spermatogenic cells were decreased,and the number of intracellular organelles including mitochondria were increased significantly in the group of Guangsiyulin Decoction.The protein levels of Bax and Bcl-2 in the group of Guangsiyulin Decoction were significantly different from those in the model group(P<0.01)through the detection of immunohistochemistry.In the Western blot and Real-Time PCR results,we found that the c-kit protein content in the testis of the Chinese medicine intervention group was higher than that in the model control group,and the corresponding mRNA expression was also significantly different(P<0.05).The protein expression of Oct4 increased,and mRNA expression also increased compared with the model control group.Although there was no significant difference(P>0.05),there was still a certain upward trend.CONCLUSIONGuangsiyulin Decoction can effectively improve the quality of semen in patients with asthenospermia,as well as various indicators of TCM symptom scores in deficiency of kidney essence,thus restore the reproductive function of patients.2.Guangsiyulin Decoction can effectively improve the general state of rat model of spermatogenic disorder,organ coefficient of genital organs,semen quality,FSH,LH,E2,T.It also can protect the morphology of testicular tissue and ultrastructure of spermatogenic cells.3.Guangsiyulin Decoction can effectively regulate the transcription and expression of Bax,Bcl-2,c-kit protein in model rats.It can obviously inhibit the apoptosis and promote the proliferation of spermatogenic cells.The expression of mRNA is not obvious,but there is a certain improvement trend.In summary,Guangsiyulin Decoction can effectively improve the male spermatogenesis,have definitely clinical efficacy,and its mechanism may be achieved by improving the serum endocrine environment and regulating the expression of Bax,Bcl-2,c-kit protein.