Experimental Study On Effects Of Kidney-nourishing Therapy On Spermatogenesis Of Mice With Kidney-Yang Deficiency Syndrome Caused By Overfatigue And Sexual Overstrain

Objective: There are many kinds of mouse models for studying kidney-Yang deficiency syndrome, but these models have certain shortcomings. To improve this situation, we developed a new mouse model for studying kidney-Yang deficiency syndrome caused by overfatigue and sexual overstrain based on classical theory of Traditional Chinese Medicine. This model more adapt to kidney-Yang deficiency syndrome in Traditional Chinese Medicine theory. The paper aims to systematic study the mechanism of " the kidney originates and controls life reproduction" targeting at spermatogenesis on tissues and organs- cytobiologic and molecular level ,so as to provide scientific basis and supports for the application of theory of "the kidney originates and controls life reproduction" in clinic treatment of male infertility by Kidney-nourishing therapy.Methods: The mouse model of kidney-Yang deficiency syndrome caused by overfatigue and sexual overstrain based on classical theory of Chinese medicine was established. The mouse were divided into three groups, normal control, model of kidney-Yang deficiency syndrome caused by overfatigue and sexual overstrain and Jinguishenqi bolus group. To determine fertility and sperm quality in each group mouse, the morphological changes of testis and quantitative assay of spermatogenesis(including the sperm density, sperm vigour and sperm abnormality rate )were observed with light microscopy; Alternation on ultramicrostructure were observed with transmission electron microscopy; gonad axis hormones (including GnRH,T,E2,FSH and LH) changes were measured by Radioimmunoassay method; the analysis of content of NO,NOS and antioxidative activity of testis was detected by cytochemical techniques; the activity of sperm-specific lactate dehydrogenase (LDH-x) in the testes was measured by cytochemistry techniques; The change of sperm-specific lactate dehydrogenase (LDH-x) gene expression was further explored by real-time quantitative reversed transcriptase polymerase chain reaction.Results: 1.The results of change of body temperature, the general condition and the picture of the tongue showed that the mouse model of kidney-Yang deficiency syndrome caused by overfatigue and sexual overstrain was established .2.In model group , the amount and motility of sperm were significantly lower than those in normal control; deformity of sperm was significantly higher than those in normal control, which reduced pregnancy rate of female and embryos fetation decrease significantly. Jinguishenqi bolus caused changes of sperm quality in male mice , and the decrease of fertility in male mice might be related to the changes of sperm quality.3. The testis index, prostate exponent, and vesicular seminalis weights of group which was treated with Jinguishenqi bolus were increased significantly.as well as,morphological structureand and Ultramcirostructural changes of testis was improved.4. The mouse model of kidney-Yang deficiency syndrome caused by overfatigue and sexual overstrain appeared dysfunction in hypothalamic-pituitary-gonadal axis, which induced spermatogenic impairment in testes. With Kidney-nourishing therapy the changes of hormone level of hypothalamic-pituitary-go nadal axis resumed.5. Jinguishenqi bolus can promote the activity of SOD, reduce the activity of NOS and the amount of NO and MDA in the testicle . Jinguishenqi bolus has the function of protecting the germ cell which is closely related to the function the the pill can provide good anti -oxidization enveriment for spermatic emergence and maturation.6. In model groups, the activity of sperm-specific lactate dehydrogenase (LDH-x) and expression of LDH-x mRNA were significantly lower than those in negative control while the conditions mentioned above increased by Kidney-nourishing therapy.Conclusion: This paper reveals that there are spermatogenesis failure and hypoxia on reproduction decline in the mouse model of kidney-Yang deficiency syndrome caused by overfatigue and sexual overstrain. With Kidney-nourishing therapy by Jinguishenqi bolus, above-mentioned function has been improved. It has provided a scientific foundation for the application of "the kidney originates and controls life reproduction" theory in clinic treatment of infertility by reinforcing kidney.