| Male infertility is a major disease that affects human reproduction and health, oligoasthenozoospermia is a very common cause. Its specific pathogenesis has not been fully clarified, modern medicine has become more and more mature in the diagnosis of oligoasthenozoospermia, but still lack of effective means in the treatment level. At present, there is still a lack of ideal therapeutic drug in clinic, and the use of drugs is empirical. Surgical treatment can be to a certain extent in orrection of organic disease and obstructive factors in male infertility. In recent years, assisted reproductive technology has developed rapidly, but because of the high cost, low success rate, genetic and ethical issues, etc. TCM syndrome differentiation and treatment has many advantages. The treatment is still much argument from the kidney, and eliminating pathogenic factors, ignoring the importance of Strengthening spleen and Supplementing Qi. Our preliminary study found that Benefit kidney and tonify spleen can significantly improve the quality of semen. According to the long-term clinical practice and combined with the theory of TCM, "the kidney is a priori, the spleen is the source of the day after tomorrow". We put forward that "deficiency of spleen and kidney essence raises" is the basic pathogenesis that oligoasthenospermia occurred, treatment should be "nourishing kidney and strengthening spleen and filling essence". At present, the research on the pathogenesis of oligoasthenozoospermia, free radical damage theory and the theory of apoptosis is a hot research topic. And mitochondrial signaling pathway is an important pathway of apoptosis. But the research on the relationship between the channel system and oligoasthenozoospermia is still not deep enough. Modern medical research shows that mitochondria are the final sites for the thorough oxidative decomposition of all matter, and its core function is synthetize adenosine triphosphate through oxidative phosphorylation and three tricarboxylic acid cycle, providing energy for the life activities of cells directly. The focus of this study is to verify YishenJianpi Recipe in the treatment of oligoasthenozoospermia, whether it is through this possible link and mechanism of action by improving the structure and function of mitochondria.Objective:To observe the effect of YishenJianpi Recipe on the testis, epididymis tissue pathology, sperm quality, serum sex hormone level, mitochondrial ultrastructure and mitochondrial membrane potential (MMP) in the mice model with oligoasthenozoospermia.Mehtod:Taking 60 male BALB/c mice, which were randomly divided into normal group, model group, positive group (Dongweili oral liquid), Chinese medicine group (YishenJianpi Recipe low-dose and high-dose group), there was 12 in each group. Mice in each group were administered intragastrically tripterygium glycosides orally for 4 weeks except the normal group (40mg/kg), as the mice model of oligoasthenozoospermia. When the model is prepared, positive group, Chinese medicine group were administered intragastrically Dongweili oral liquid (10ml/kg), YishenJianpi Recipe low-dose group (1.35mg/kg) and high-dose group (2.70mg/kg) orally for 4 weeks, normal group and positive group were administered intragastrically Equivalent distilled water orally. After the end of treatment, pathological section HE staining and electron microscope were used to observe the pathological changes of testis and epididymis tissue in mice; Weili sperm tester were used to observe the semen quality in mice; the level of sperm mitochondrial membrane potential (MMP) was detected by applying JC-1 fluorescence staining flow cytometry; detection of serum sex hormone levels in mice by enzyme linked immunosorbent assay; the ultrastructure of mitochondria was observed by transmission electron microscope.Results:1 pathological of testis and epididymis:In the normal group, the testis of the mice is fine and precise, which is closely arranged, it contains all levels of germ cells and Sertoli cells. In model group, the testis of mice was atrophy, all levels of germ cell arrangement were disorder and loose, the quantity was decreased obviously, and the Sertoli cells were more serious. After drug intervention, a few cells in the small number of the fine sperm cells was decreased, and the degree and range of the lesions were significantly reduced. In normal group the epididymal lumen stromal tissue enriched orderly, the luminal epithelial cells clearly arranged neatly in a tall columnar, showing that a large number of sperm. In the model group, there was a decrease of interstitial tissue and interstitial tissue, and the epithelial cells arranged in the cavity were disordered or absent. After drug intervention, compared with the model group, the outer tissue of epididymis was more ordered, the number of epithelial cells increased, the arrangement was still neat, the structure was almost complete.2 semen quality:Compared with model group, A levwl, (A+B) level and total motility in positive group and Chinese medicine low-dose and high-dose group were higher, the difference was statistically significant (P<0.05); Compared with positive group and high-dose group, low-dose group was higher, the difference was statistically significant (P<0.05); There was no statistical significance between positive and high-dose group (P value is 0.4030, 0.4676,0.4405, P>0.05); Compared with model group, sperm concentmiceion and total in normal group, positive group and Chinese medicine low-dose and high-dose group were higher, the difference was statistically significant (P<0.05); there was no statistical significance among positive, low-dose and high-dose group (P>0.05)3 mitochondrial structure and membrane potential of spermatozoa:Compare with the normal group, MMP in the model group was lower, the difference was statistically significant (P<0.0001); compared with the positive and low-dose group, MMP in the high-dose group was higher, the difference was statistically significant (P value is 0.0008,0.0001, P<0.05); compared with the positive group, MMP in the low-dose group was lower, the difference was statistically significant (P<0.05). In normal group, the middle section of the mice sperm tail showed the complete mitochondrial sheath structure, ring wrapped in a dense fibrous and axial filament peripheral 9 fiber,9 root axis wire and two structure clear microtubules, together constitute the structure of 9+9+2. In model group, mitochondrial swelling, loose fiber, and axial filament were broken, and the structure of 9+9+2 was destroyed. After drug intervention, the sperm mitochondrial sheath intact, mitochondrial swelling was significantly reduced compared with the previous, dense fiber, axial filament structure is neat and complete, 9+9+2 structure is clear.4 serum sex hormone level:Compare with the normal group, serum follicle stimulating hormone and lutein level in other group were higher, the difference was statistically significant (P<0.05); There was no difference in serum follicle stimulating hormone and lutein level between model group and each intervention groups (P>0.05). Compared with model group, each intervention group was lower, the difference was statistically significant (P<0.05). Compared with normal group, the serum T level in model group was lower, the difference was statistically significant (P=0.0008<0.05). There was no difference in serum T level between model group and each intervention groups (P>0.05)Conclusion:YishenJianpi Recipe can regulate serum sex hormone level of mice of oligoasthenotspermia induced by tripterygium glycosides; reduce the pathological damage of the testis and epididymis of mice; improve the structure of sperm mitochondria and the mitochondrial membrane potential; improve spermatogenesis and mature, so as to improve the quality of semen in the mice of oligoasthenozoospermia. |
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