The Effect Of Jinguishenqi Pill On Kidney Yang Deficiency Of Spermatogenesis In Rat And TGF β1/CYP19

In recent years, male reproductive health is increasingly under threat, the malesterility rate has increased from3:7to5:5, male spermatogenesis dysfunction has beenpaid more and more attention. spermatogenesis refers to the formation of sperm cells fromthe original continuous proliferation, differentiation, development process, experiencedspermatogonia proliferation, spermatocyte meiosis and spermiogenesis in three stage [1].Spermatogenesis is relates to the interaction of spermatogenic cells, Leydig Cells andSertoli Cells between the supporting cells, and regulation of [2-5] precision adjustmentand many genes Luteinizing hormones, Follicle Stimulating Hormone, Androgen andendogenous sex hormone estrogen. Many previous studies focused on LH, FSH, ANG, andto pay less attention to the endogenous estrogen, but estrogen has the important role in theregulation of male sperm production, the main source of endogenous estrogen in male body is composed of androgen in aromatase role to the conversion, and the CYP19geneencodes aromatase. Studies have found that [6-11], CYP19through the regulation of thesynthesis of spermatogenesis in the testis of regulation of estrogen, Transforming growthfactor beta1can inhibit the expression of CYP19and rigid sperm production, also canaffect sperm production."Reproduction" is the highly generalization of traditional Chinese medicine onhuman reproductive function, men, including sperm production depends on the Yinnourishing and kidney warming, number depends mainly on the kidney essence and Qi ofyin and yang. Chinese medicine, kidney deficiency is the cause of spermatogenic failure,the leading cause of male infertility. Jinkui Shenqi Pill recorded in "synopsis ofprescriptions of the Golden Chamber of Zhang Zhongjing", is based on the six tastesglutinous rehmannia pill plus Ramulus Cinnamomi, Radix Aconiti Lateralis Preparata andbecome, Jinkui Shenqi Pill as tonic kidney-yang recipe classic, which has a significanteffect on the reproductive system of male rats, which can not only improve the kidneydeficiency caused by the sperm count and motility rate [12], but also can improve thespermatogenic function, but so far, improve the mechanism of deficiency of kidney-yanginfertility mainly by regulating the hypothalamic-pituitary-gonadal axis in the functionof promoting the improvement of spermatogenic function of Jinkui Shenqi pill. But, on theexpression of TGF-β1/CYP19rarely reported, therefore, in order to further clarify itsmechanism of action.The experiment is divided into three parts:Section1The effect of Jinkuishenqipill onSpermatogenesis of rat suffering kidney-yang deficiency andexpression of TGF-β1/CYP19Objective: Observing Jinkuishenqipill’s effect on adenine-induced infertile rat caused bykidney yang from changes of blood cleaning hormone level, sperm concentration, spermmotility and TGF-β1/CYP19expression.Mdthods:30adult male SD rats were randomly divided into three groups: normal control group, model group and treatment group (10per group). After successful adeninemodeling method, use13.5g/kg d of Jinkuishenqipill to gavage rats in treatment group for30days. Then oberve the situation of rats in every group. Weigh all rats after modeling.Use sex hormone radioimunoassay kit to check the level of sex hormones in all rats. Useautomatical sperm detector to test the density and motility of sperm. Useimmunohistochemistry to detect the expression level of TGF-β1/CYP19in rats testis.Results: After modeling, the weight of rats in model group was significantly reduced（P﹤0.05）, compared with rats in the control group. Compared with normal control group, Tand E2were significantly reduced (P<0.05). FSH and LH had no significant changes(P>0.05). Sperm density was reduced（P﹤0.05）. Sperm motility was reduced（P﹤0.05）.The immunohistochemical result shows that TGF-β1and CYP19were expressed intestis long sperm cells, expression levels of TGF-β model of rats in the1groups wassignificantly higher than that in control group, while the Jinkuishenqiwan treatment, itsexpression decreased slightly, but the Jinkuishenqiwan treatment group, TGF-β1expression is still higher than the normal control group, the expression of CYP19in therats of model group is significantly lower than that of normal control group, while theexpression of Jinkui Shenqi Pill in the treatment group was higher than that of the modelgroup and normal control group.Conclusion: Jinkuishenqipill can increase the weight, serum sex hormone levels andsperm density and motility of rats with deficiency, reducing the expression of TGF-β1inrats testis and increasing the expression of CYP19.Section2Spermatogenesis and expression of TGF-β1/CYP19of rats withkidney yang after preventing TGFβRⅠObjective: Observing sex hormone levels, sperm density and motility andTGF-β1/CYP19expression of rats after microinjection.Mdthods:40adult male SD rats were randomly divided into four groups: normal controlgroup, normal+blocker group and model+blocker group (10per group).20adult male SD rats were microinjected with TGFβRⅠ to prevent TGF-β1expression. Check sexhormone levels in rats. Use automatic sperm detector to check the sperm density andmotility. Use immunohistochemistry to detect the expression level of TGF-β1/CYP19inrats testis.Results: after microinjection, compared normal+blocker group and model+blocker groupwith normal control group, serum T and E2increased. Sperm density and motilityincreased. They were statistically significant (P<0.05). Immunohistochemical results showthat, positive expression of TGF-β1, located in long sperm cells, the positive expressionof CYP19located in long sperm cells and round spermatids, block rat testis expression ofTGF-β1was significantly decreased, while CYP19expression increased significantl.Conclusion: Microinjection of TGFβRⅠ blockers, the number of testicular sperm ratsincreased, sperm survival rate than those in the model group were improved.Section3Jinkuishenqipill’s effect on Spermatogenesis of rats withkidney yang and TGFβRⅠand TGF-β1/CYP19Objective: Observing Jinkuishenqipill’s effect on serum sex hormones and sperm density,sperm motility and TGF-β1/CYP19expression of rats after microinjection.Mdthods:60adult male SD rats were randomly divided into normal group, normal+treatment group, model group, model+treatment group, block group, block and treatmentgroup,(10per group).10adult male SD rats were treated by Jinkuishenqipill aftermicroinjection of TGFβRⅠ.30days later, kill the rats. Serum, semen and testicular tissueare kept. Check serum sex hormone level of all rats. Use automatic sperm detector tocheck sperm density and motility. Use immunohistochemical method to checkTGF-β1/CYP19expression in rats testis.Results: The model group and model plus treatment group, block group and blocktreatment group， there is no difference among serum sex hormone level, sperm densityand motility (P>0.05). immunohistochemical method results show that, positiveexpression of TGF-β1/and CYP19in elongating spermatids, two groups of rats with TGF-β1/and CYP19expression were of no significant difference, but compared with thenormal group rats compared with expression of TGF-β1, is still high, but the expressionof CYP19is still lower.Conclusion: Jinkui Shenqi Pill could obviously improve the rat model of kidney yangdeficiency symptoms, increase sperm production, its mechanism is possibly throughinhibiting the expression of TGFβRⅠ, thereby affecting the sperm production.Providesthe reliable experimental basis for further explore the mechanism of treatment of malereproductive disorders of Jinguishenqi pill.