| Objective: study the therapeutic effects of the alcohol extract,water extract and extract composition of Yimusake on the changes in sexual function of ED rats and the key indicators such as e NOS,n NOS,VIP,ET-1 in the gonadal axis and penis tissue of ED rats,evaluate the drug efficacy and screen its active ingredients,and explore its possible mechanism.Methods:(1)160 male SD rats with normal sexual function were randomly divided into a normal control group(group N,15 rats)and a modeling group(group M,145rats).The N group received standard diets and a regular environment(22±2℃,humidity55～65%);Group M is fed with special estrogen-like feed in a cold environment(4±0.5℃,humidity 85-95%,between 10:00-18:00 daily).Dynamically observe the changes in biological signs and sexual performance of rats in each group.After 35 weeks,ED rats were screened from the group M and randomly divided into an erectile dysfunction group(group ED,12 rats),an alcohol extract group(group AE,12 rats),a water extract group(group WE,12 rats),the extract composition group(group COE,12 rats),and the Yimusake tablet control group(group Y,12 rats),respectively,with distilled water,alcohol extract,water extract,alcohol extract and water extract The composition and Yimusake tablets were intervened for 3 weeks.Dynamically observe the changes in sexual behavior of rats in each group,and evaluate the drug effects.(2)Detect the changes of serum gonadal axis hormones in each group of rats by radioimmunoassay,stain the tissues of hypothalamus,pituitary and testis with HE,and observe the morphological changes under a light microscope.(3)Detect the expression of e NOS,n NOS,VIP and ET-1 in the penile tissues of rats in each group.Results:(1)Modeling factors Changes in the biological signs of rats in each group during the intervention: There was no significant difference in the weight,diet,water intake,urine output,and stool volume of the rats in each group before the intervention(P>0.05).In the first 13 weeks after the intervention of the modeling factors,compared with group N,the weight of group M increased significantly(P<0.05).From the 5th week of modeling,the amount of food,urine,and stool in group M were significantly higher than that in group N(P<0.05).From the 13 th week,the amount of drinking water was significantly reduced in group M(P<0.05).(2)Changes in the sexual behavior of rats in each group during the intervention of modeling factors: Before intervention,the latency and times of crawling back,penetration,and ejaculation in group M were not significantly different from those in group N(P>0.05);from the 9th week to the end of modeling In group M,the number of crawling back,penetration,and ejaculation were significantly reduced(P<0.05);the latency of crawling back,penetration and ejaculation was significantly prolonged(P<0.05).(3)Changes in sexual behavior of rats in each group after the intervention of Yimusake extract and composition: After one week of intervention,the latencies of crawling back,penetration,and ejaculation in group ED,AE and WE were significantly longer than those in group N(P<0.05),the number of back crawling,insertion,and ejaculation were all less than that of group N(P<0.05).After 2 weeks of intervention,the latency of back crawling,penetration and ejaculation in group ED and WE was significantly longer than that in group N(P<0.05).The number of back crawling,penetration,and ejaculation in group ED were significantly less than those in group N(P<0.05),and there was no statistically significant difference between the intervention groups and group ED(P>0.05).After 3 weeks of intervention,the latency of back crawling,penetration,and ejaculation in group ED was significantly longer than that in group N(P<0.05).The number of climbing backs in group WE and COE was higher than that in group ED(P<0.05).The insertion latency of group WE and COE was shorter than that of group ED(P<0.05).The number of insertions in group COE was higher than that in group ED(P<0.05).The ejaculation latency of group WE and COE was shorter than that of Group ED(P<0.05).The times of ejaculation in group WE and COE were higher than those in group ED(P<0.05);compared with group N,there was no significant difference between group AE,WE and COE in the latency of back crawling,insertion latency,back crawling,and the number of insertions(P> 0.05).The latencies of back crawling,penetration,and ejaculation in group AE were significantly longer than those in group N(P<0.05),and the number of ejaculation in group AE was less than that in group N(P<0.05).There was no significant difference in the ejaculation latency and the number of ejaculation between group WE and COE and group N(P>0.05);compared with group Y,group AE,WE and COE had no significant difference in back-climbing,penetration,ejaculation latency and frequency Sexual difference(P>0.05).(4)The effect of Imusak extract and composition on the changes of gonad axis hormones in ED rats: Compared with group N,the testosterone level of group ED was significantly reduced(P<0.05),and group COE testosterone level was significantly increased(P<0.05)),the estradiol level in group AE was significantly reduced(P<0.05);compared with group ED,the testosterone level in group COE was significantly increased(P<0.05);there was no significant difference between the other groups(P>0.05).There was no statistical difference in LH,FSH,PRL among the groups(P>0.05).(5)The effect of the extract and the composition on the morphological changes of the gonadal axis in ED rats: the structure of the hypothalamus,pituitary gland,and testis in group N was normal.Compared with group N,the hypothalamic paraventricular nucleus nerve cells proliferated and arranged disorderly in group ED;the paraventricular nucleus nerve cells of group AE,WE,COE and Y were repaired,and the cell arrangement was more regular than that of the group ED.In group ED,the arrangement of adenohypophysial cells was disordered,with tissue edema,and vacuole-like changes were more common;cell edema and vacuole-like changes in group AE,WE,COE and Y were improved.Sperm can be seen in the seminiferous tubules of the testis in group ED,but it is not as full as group N,the interstitium is obviously congested and edema,and the number of supporting cells is significantly reduced;the number of sperm in the seminiferous tubules of the testis in group AE,WE,COE and Y increased,and The hyperemia was improved,and the number of supporting cells increased compared with group ED.(6)The effect of Imusak extract and composition on the expression of e NOS,n NOS,VIP,ET-1 in ED rats: The results of immunohistochemistry and Western-blot showed that e NOS was expressed in the penile tissues of rats in each group.It is mainly expressed in the cytoplasm of the penile cavernous blood vessels and cavernous sinus endothelial cells,as well as in the nucleus.The expression of e NOS in group ED was significantly lower than that in group N(P<0.05);the expression of e NOS in the AE,WE group,and COE was significantly higher than that in group ED(P<0.05),compared with the Y group There is no significant difference(P>0.05).n NOS was expressed in the penile tissues of rats in each group,mainly in the cytoplasm of cavernous smooth muscle and vascular wall,cavernous sinus endothelial cells,and also in the nucleus.Compared with group N,the expression of n NOS in group ED was significantly reduced(P<0.05),and group AE,COE,and group Y all increased to different degrees(P<0.05);compared with group ED,The expression of n NOS in group COE and Y were significantly increased(P<0.05);compared with group Y,the expression level of n NOS in group WE was decreased(P<0.05),and the expression level of n NOS in group AE and COE was lower than that in group Y.There was no significant difference in group Y(P<0.05).VIP was expressed in the penile tissues of rats in each group,mainly in the nucleus of the smooth muscle of the corpus cavernosum and vascular endothelial cells,and also expressed in the cytoplasm.Compared with group N,the expression of VIP in group ED was significantly reduced(P<0.05),and the expression of VIP in group WE,COE and Y was significantly increased(P<0.05);compared with the group ED,the AE The expression of VIP in group WE,COE and Y were all significantly increased(P<0.05);compared with group Y,the expression of VIP in group COE was significantly increased(P<0.05).ET-1 was expressed in the penile tissues of each group of rats,mainly expressed in the nucleus and cytoplasm of the corpus cavernosum endothelium and the endothelial cells of the cavernous sinus.Compared with group N,the ET-1 expression in group ED was significantly increased(P<0.05).Compared with group ED,the expression of ET-1 in WE and COEs was significantly lower(P<0.05).Compared with group Y,the expression of ET-1 in COE and WEs were significantly reduced(P<0.05).Conclusion:(1)after 35 weeks of environmental estrogen like diet combined with cold stimulation,the modeling rate of ED was 75.90%,and the model was established successfully.(2)The results showed that both the water extract and the extract composition of imasak could significantly improve the sexual function of rats,and there was no significant difference between the effect of imasak and that of imasak tablets.The expression of e NOS,n NOS,VIP and ET-1 in penile tissue of ED rats is one of the important pathogenesis of ED.(3)The results suggest that COE may improve the sexual function of ED rats and play a therapeutic role by increasing the serum testosterone level.(4)We and COE upregulate e NOS,n NOS,VIP and downregulate ET-1,which may be one of the important mechanisms of we and Coe’s therapeutic effect. |
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