Clinical And Experimental Study On The Treatment Of Infertility Of Spermatozoa With Weak Sperm In

In recent years, with the improvement of people’s living standard, traditional way of life has undergone tremendous changes. These changes led to many negative factors, such as overeating, anxiety and long-term stay up late, work overload and other factors directly lead to male sperm levels declining, men fertility suffer huge challenge. How to effectively improve male fertility has become a hot topic of the moment. However, the current status of clinical is that there is not effectively treatment of male infertility in Western medicine, which is to rule out assisted reproductive technologies, infection, obstruction and other clear causes. At the same time, assisted reproductive technology can help solve fertility problems in some patients, but long-term effects on genetic problems remain to be seen. In order to promote the study of male infertility, the majority of clinicians are turning to Traditional Chinese Medicine for the diagnosis and treatment of male infertility. Male infertility which is treated by Traditional Chinese Medicine has a long history and significant clinical efficacy. In this case, how to fully exploit the advantages of Chinese medicine treatment of male infertility has become an important issue.The purpose is to evaluates clinical curative effect of oligoasthenotspermia of kidney yin deficiency type by using Zuo Gui Pill of this article; Then to explore reasonably oligoasthenotspermia type rat model which is prepared by Tripterygium glycosides; After above, protection and treatment experiment were been designed in pharmacodynamic study to comprehensive evaluate the therapeutic effect of organ coefficient rats, semen quality, sex hormone levels, epididymal carnitine, inhibin B levels by using Zuo Gui Pill; Finally, in the protection and treatment experiment, each group testis was tested to Bax, Bal-2, SCF, Ki67 protein and mRNA expression levels used by immunohistochemistry, western blot and Real-Time PCR technology to research anti-apoptotic and proliferation mechanism of Zuo Gui Pill by qualitative and quantitative analysis.Part oneThe clinical research for the effect in treating oligoasthenotspermia of male infertility by Zuo Gui PillObjective:To research the clinical efficacy in treating oligoasthenotspermia of kidney yin deficiency type by Zuo Gui Pill.Methods:In this study, before and after self control study was used. Treatment period was 90 days,42 patients with oligoasthenotspermia of kidney yin deficiency type were given Zuo Gui Pill formula granules for treatment. Routine examination of semen was performed once before and after treatment in patients; After the start of the research, patients should been follow-up every 30 days with TCM syndrome score; At the same time concomitant medications, adverse events, pregnant or not were recorded. To evaluate improvement of semen quality and TCM syndromes on oligoasthenotspermia of kidney yin deficiency type treated.Results:Compared with before treatment, semen liquefaction state, semen volume, a grade sperm percentage, b grade sperm percentage, the percentage of non-forward motility, total sperm motility, sperm concentration, total sperm of after treatment were improved significantly, there was statistically significant (p<0.01); The total score of TCM Syndrome and score of kidney yin deficiency were associated with the number and the time of follow-up, which was showed a downward trend, there was statistically significant (p<0.01). During the study, there was not adverse reaction report.Conclusion:Zuo Gui Pill was safe and effective in improving sperm quality on oligoasthenotspermia of kidney yin deficiency type, as well as improving the patient’s TCM symptoms, particularly significant improving kidney yin deficiency symptoms.Part twoEstablishing reasonably oligoasthenotspermia type rat model by Tripterygium glycosidesObjective:To research several different group of Tripterygium glycosides dosage and modeling time; To observe rat model recovery; To establish relatively reasonable oligoasthenotspermia type rat model.Methods:96 male SD rats were randomly divided into normal group, Tripterygium glycosides high dose group, middle dose group and low dose group. In the normal group, deionized water was used to intervene; Tripterygium glycosides high dose group, middle dose group and low dose group were used 50 mg/(Kg·d),40 mg/(Kg·d),30mg/(Kg·d) of Tripterygium glycosides to intervene. The 3th and 4th week, four SD rats were selected to execute from each group; After the 4th week, four SD rats were selected with the above experimental program feeding, and were executed in the 5th week; Remaining rats were stopped to intervene. And four SD rats were selected in the 6th week, the 7th week, and the 8th week to observed spermatogenesis recovery.Results:During the modeling period (the 3th week to the 5th week), general condition, genital organ coefficient, semen quality indicators was significantly reduced tendency along with Tripterygium glycosides dosage increase and extend the treatment time; Testicular pathological injury was significantly increased trend along with Tripterygium glycosides dosage increase and extend the treatment time. After stopping the treatment (the 6th week to the 8th week), indicators of Tripterygium glycosides low dose group gradually returned to normal levels; Indicators of Tripterygium glycosides middle dose group recovered slightly, but compared with the normal group, there still was a wide gap; There was no significant damage restoration in the high dose group.Conclusion:Considering the modeling time, the economic costs and rats self-recovery, reasonable oligoasthenotspermia type rat model could be established by using 40mg/(Kg·d) of Tripterygium glycosides to intervene 4 weeks.Part threePharmacodynamic Study on oligoasthenotspermia type rat model by using Zuo Gui PillObjective:To observe the improvement of general condition, organ coefficient, semen quality, pathological changes in testis, sex hormone levels, epididymal carnitine, serum inhibin B of oligoasthenotspermia type rat model by using Zuo Gui Pill.Methods:72 male SD rats were randomly divided into treatment and protection experiments, and each experiment was divided into normal group, model group, Zuo Gui Pill group. In the protection experiment, the normal group was used deionized water to intervene four weeks; The model group was been made the model with 40mg/(Kg"d) of Tripterygium glycosides four weeks; The Zuo Gui Pill treatment group was been made the model with 40mg/(Kg"d) of Tripterygium glycosides four weeks, at the same time with 6g/(Kg"d) of Zuo Gui Pill suspension (equivalent to 20 times the human drug) interventions to protect 4 weeks. In the treatment experiments, the normal group was used deionized water 8 weeks; The model group was been made the model with 40mg/(Kg·d) of Tripterygium glycosides four weeks, from the 5th week to 8th week deionized water was given to intervene; The Zuo Gui Pill treatment group was been made the model with 40mg/(Kg·d) of Tripterygium glycosides four weeks, from the 5th week to the 8th week 6g/(Kg·d) Zuogui pills suspension (equivalent to 20 times the human medication) was given to intervene.Results:In the protection experiment, compared with the model group, Zuo Gui Pill group rat testis organ coefficient epididymis organ coefficient, seminal vesicles organ coefficient, the indicators of semen quality, testosterone levels, serum inhibin B were differences, there were statistically significant (p<0.01or p<0.05); In the treatment experiments, compared with the model group, Zuo Gui Pill group rat testis organ coefficient, organ coefficient epididymis, prostate organ coefficient, semen quality indicators, testosterone levels, serum inhibin B were differences, p<0.01or p<0.05, there were statistically significant; In the protection and treatment experiment, compared with the model group, Zuo Gui Pill group general condition, testicular tissue pathology obviously alleviated.Conclusion:the general condition, genital organ coefficient, semen quality, testosterone levels, serum levels of inhibin B of rat model could be effectively improved by using Zuo Gui Pill, there was obvious protective and treatment effects.Part fourInfluence on apoptosis and SCF, Ki67 protein and mRNA expression in oligoasthenotspermia type rat model by using Zuo Gui PillObjective:To investigate influence on apoptosis regulatory factors Bax, Bcl-2 protein and mRNA in oligoasthenotspermia type rat model by using Zuo Gui Pill; at the same time,to observe the effect on SCF, Ki67 protein and mRNA expression by using Zuo Gui Pill.Methods:By Immunohistochemistry, western blot and Real-Time PCR technology, all experimental rats testis tissue in the treatment and protection experiment of Zuo Gui Pill pharmacodynamic studies were been tested with Bax, Bcl-2, SCF, Ki67 protein and mRNA expression.Results:In the protection experiments, with immunohistochemistry technology, compared with the model group, Zuo Gui Pill group Bax, Bcl-2, SCF, Ki67 protein were significant differences, P<0.05 or P<0.01, there was statistically significant; With western blot technology, compared with the model group, Zuo Gui Pill group Bax, Bcl-2, SCF protein were significant differences, P<0.05, there was statistically significant; With Real-Time PCR technology, compared with the model group, Zuo Gui Pill group Bax, Bcl-2, SCF, Ki67 mRNA were obvious differences, P<0.05 or P<0.01, there was statistically significant. In the treatment experiments, with immunohistochemistry technology, compared with the model group, Zuo Gui Pill group Bcl-2,SCF,Ki67 protein were significant differences, P<0.05 or P <0.01, there was statistically significant; With western blot technology, Zuo Gui Pill compared with the model group, there was no significant difference, but Bcl-2 and SCF expression was significantly higher than the model group; With Real-Time PCR technology, compared with the model group, Zuo Gui Pill group SCF, Ki67 mRNA had a significant difference, P<0.05 or P<0.01, there was statistically significant.Conclusion:These data indicate that Bcl-2, SCF, Ki67 protein and mRNA expression are been increased by using Zuo Gui Pill, as well as Bax protein and mRNA expression is been reduced. It shows that Zuo Gui Pill has the significant function to inhibit spermatogenic cell apoptosis and promote spermatogenic cell proliferation.