| Varicocele(V C)is an angiopathy that refers to the abnormal expansion,elongation,and distortion of the plexus within the spermatic cord,which can lead to pain discomfort and progressive testicular hypofunction.It is one of the common incentives of male infertility.Recent studies have shown that the regulation of reproductive hormones and the effect of apoptosis on spermatogenic cells is an important factor in the induction of spermatogenesis by VC.The clinical use of Tongmai decoction in treating VC-infertility received satistify results,can effectively improve the quality of semen caused by VC decline.To further explore the mechanism of Tongmai decoction in trating VC,carry out this experimental study.Objective:To investigate the regulation of Tongmai decoction on serum hormone levels and the regulation of Spermatogenic cell apoptosis in VC rats,and to evaluate the effect of tongmai decoction on the function of testicular spermatogenic in VC model rats.Methods:72 7-week SD male rats were randomly divided into 12 rats of blank control group and 60 models rats,and the model of VC was established by reference to Turner's classical method,ligation of gonadal veins by microsurgical instruments,and reduction of collateral circulation.After 30d,the rats were randomly divided into 5 groups:Model control group,aescuven group,Tongmai decoction low-dose group,Tongmai decoction group and Tongmai decoction High dose group,each group 12 rats.After grouping,the rats were fed Drug through stomach,and the volume of the drug was 1ml/100g weight.In the aescuven group,the rats were fed water solution of aescuven,the dosage was 5.4mg/kg body weight,equivalent to the clinical dosage,and the Tongmai decoction was administered to the rats with Tong Mai decoction,and the dosage was 5.45g/kg body weight,which was equivalent to half of the clinical dosage.Tongmai Decoction was administered to the rats in the dose group,the dosage was 10.89g/kg body weight,which was equivalent to the clinical dosage,and the rats were given Tongmai decoction,the dosage was 21.78g/kg,and the weight was twice times of the dosage of clinical medicine.In the model control group,the blank Control group,the volume of distilled water,such as the upper method,was given once a day,with a total of 48d.After 48 days of drug intervention,the rats were anesthetized and the blood was collected from the inferior vena cava.Rat serum luteinizing hormone(LH),follicle stimulating hormone(FSH),and testosterone(T)concentrations were measured by radioimmunoassay.The bilateral epididymis tissue was smashed and the semen was released.The rat semen was analyzed using the WLJY-9000 Beijing Weili Color Sperm Quality Assay System.The testes were cut into two halves,half were HE stained for pathological changes and immunohistochemical detection of Bax,Bcl-2,Hsp60,Hsp90 expression,the other half was used for Western Blot detection of Bax,Bcl-2,Hsp60,Hsp90 expression.Result(1)HE staining:In the blank control group,the boundary of the seminiferous tubules was neat,and there was no shrinkage and collapse.There was no hyperplasia in the interstitial cells,and tight connections between the seminiferous tubules were found and the basement membrane was intact.The supporting structure was abundant,and a large number of mature spermatozoa were seen in the lumen.Model control group:The testicular tissue showed disordered arrangement of seminiferous tubules,degenerative spermatogenic epithelial cells,rare sperm;interstitial edema,interstitial cell hyperplasia;interstitial small blood vessels congestion,thickening of the wall,lumen stenosis;Tongmai detection low-dose group:the arrangement of seminiferous tubules was orderly and regular,and the size was relatively uniform;a few spermatogonia epithelial cells were degenerated and the spermatozoa could be seen;interstitial cell hyperplasia,interstitial edema,and occasional hyperemia of small interstitial blood vessels;Tongmai detection middle dose group:the seminiferous tubules are ordered and arranged in order,the borders are neat,visible at all levels of spermatogenic epithelial cells,spermatozoa can be seen;rare interstitial cell proliferation and interstitial edema,wall thickening is not obvious;Tongmai detection high-dose group:The seminiferous tubules were arranged in a regular and orderly manner,and the size was relatively uniform.Seen at all levels of seminiferous epithelial cells,there were a large number of spermatozoa in the lumen;rare interstitial edema and interstitial cell hyperplasia were rare;The spirit of the aescuven group:The seminiferous tubules are arranged in a regular and orderly manner,the size is relatively uniform,and spermatozoa are rare;no interstitial edema and no interstitial cell hyperplasia are seen.(2)The sperm concentration analysis showed that compared with the control group,the sperm concentration in the model control group was significantly decreased(P<0.05).Compared with the model control group,the sperm concentration in the Tongmai detection dose group was significantly improved(P<0.05).(3)Sperm total vitality analysis showed that compared with the blank control group,the total sperm viability of the model control group was significantly lower(P<0.05);compared with the model control group,the total sperm viability of the Tongmai detection dose group was significantly increased(P<0.05).),The total sperm viability of the aescuven group was not significantly improved(P>0.05).It indicates that tongmai side can improve the total sperm vitality,the effect is better than aescuven.(4)The serum FSH test showed that compared with the control group,the FSH secretion in the model control group increased significantly(P<0.05);compared with the model control group,the FSH secretion in the low-dose Tongmai detection group and the middle-dose Tongmai detection group significantly decreased(P<0.05).(5)The test of Serum LH showed that compared with the control group,the LH secretion in the model control group was significantly higher(P<0.05);compared with the model control group,the LH secretion was significantly decreased in the Tongmaifang dose group(P<0.05).(6)The test of Serum T showed that compared with the blank control group,T secretion in the model control group decreased significantly(P<0.05).Compared with the model control group,T secretion of Tongmai detection side dose group was significantly higher(P<0.05).(7)The test of Hsp60 testis showed that compared with the blank control group,the expression of Hsp60 in the model control group was significantly decreased(P<0.05);compared with the model control group,the expression of Hsp60 in the Tongmai detection dose group and aescuven group was significantly higher(P<0.05),indicating that both doses Tongmai detection and aescuven can up-regulate Hsp60.(8)Hsp90 testis test showed that compared with the blank control group,the expression of Hsp90 in the model control group was significantly decreased(P<0.05);compared with the model control group,the expression of Hsp90 was significantly increased in each dose group and aescuven group(P<0.05),indicating that both doses Tongmai detection and aescuven can up-regulate the expression of Hsp90.(9)The test of Bax showed that compared with the control group,the expression of Bax in the model control group was significantly increased(P<0.05);compared with the model control group,Bax expression was significantly decreased in each group of Tongmai detection and aescuven group(P<0.05),indicating that Tongmai detection and aescuven can reduce Bax expression in each dose group.(10)The test of Bcl-2 showed that the expression of Bcl-2 in the model control group was significantly lower than that in the control group(P<0.05).Compared with the model control group,tongmai detection dose group and aescuven group were larger than the control group.The expression of Bcl-2 in rat testis tissue was significantly increased(P<0.05),indicating that both Tongmai detection and aescuven can significantly up-regulate Bcl-2 expression in each dose group.Conclusion:The mechanism of Tongmai decoction in repairing spermatogenesis in VC rats may inhibit the apoptosis of spermatogenic cells in testis.The mechanism of inhibiting apoptosis of spermatogenic cells involves up-regulating Hsp60,up-regulating Hsp90,down-regulating Bax,and up-regulating Bcl-2 expression.Regulate the secretion of reproductive hormones,thereby protecting the testicular spermatogenesis and improving sperm quality.Tongmai decoction can significantly improve sperm quality,and its efficacy is significantly better than that of aescuven. |
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