| Objective:Male oligoasthenospermia is one of the important causes of male infertility.About 90%of infertile men have the problem of decreased sperm count and sperm motility,which has become a social hot issue affecting male reproductive health.At present,western medicine has some limitations in the treatment of this disease.A considerable number of sperm dysfunction can not be well solved.Traditional Chinese medicine has its own unique treatment methods in the treatment of this disease and has achieved significant treatment results,which can make up for the deficiency of Western medicine in the treatment of male infertility.The research group has explored that microwave radiation is one of the causes of oligoasthenospermia.The clinical investigation results show that kidney yang deficiency syndrome is the main syndrome type of traditional Chinese medicine.Through the mechanism research,it is found that Jinkui Shenqi Pill can improve the microenvironment of spermatogonial stem cells damaged by microwave radiation and regulate the self-renewal and differentiation of spermatogonial stem cells.In this study,a retrospective study will be conducted to explore the use of kidney tonifying drugs in male infertility.At the same time,through experimental research,we explored the relationship between the mechanism of Hailong Huangjing powder in the Spermatogenesis Function of mice with oligoasthenospermia induced by microwave radiation and the cytokines,oxidative stress,and apoptosis,and explored the role of acupuncture and moxibustion in this disease.Methods:Clinical research:Based on the ancient and modern medical case cloud platform,through the method of retrospective research,use the statistics,cluster analysis,and association analysis in the platform to mine the drug use rules and core prescriptions of syndromes.Experimental study:Balb/c male mice were randomly divided into the group A as blank group,group B as microwave radiation model group,group C,D,E as radiation with Hailong Huangjing Powder group 1,2,3,and group F as acupuncture group,with 8 mice in each group.Group B,C,D,E,and F were treated with 2450MHz continuous microwave with the power density of 40mw/cm~2 for 18 days and 1H per day.Group C,D,and E were given 0.0075g,0.015g,and 0.03g of Hailong Huangjing powder by gavage every day.Group F was given"Sanyinjiao(SP6)" by hand acupuncture every day for 18 days.After the intervention,sperm motility,the staining sections of testes,and the expression of Bax,Bcl-2,and Caspase-3 in testes were detected by immunofluorescence;Detection of TNF-α,IL-6 and IL-12 in testicular tissue by ELISA;Superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH_Px)and malondialdehyde(MDA)were detected by biochemical methods.Results:Clinical study:1434 prescriptions were included in this study,including 1434 prescriptions and 186 traditional Chinese medicines.The frequency of drug appearance was 17472 times in total.Among them,dodder,Schisandra Chinensis,licorice,medlar,Cinnamon Twig,plantain seed,tuckahoe,white peony,Angelica Sinensis,and raspberry are used most frequently.According to the association rules,the core traditional Chinese medicine of kidney deficiency syndrome is Schisandra Chinensis,dodder,medlar,and plantain seed;The core traditional Chinese medicine of kidney deficiency and stasis syndrome is Ramulus Cinnamomi,licorice,dodder,Angelica Sinensis and Schisandra Chinensis;The core traditional Chinese medicine of spleen kidney deficiency syndrome is Schisandra Chinensis,dodder seed,tuckahoe,and licorice.Experimental study 1:compared with group A,after microwave irradiation,Sertoli cells in seminiferous tubules of mice in group B were destroyed,spermatogenesis was dysfunctional,and sperm cells in seminiferous tubules decreased.The levels of PR,NP,and PR+NP in spermatozoa were lower than those in group A(P<0.05).Compared with group B,the sperm NP level of group C was higher than that of group B(P<0.05),and the sperm PR,NP,PR+NP level of group D was higher than that of group B(P<0.05),the sperm PR,NP,PR+NP level of group E was higher than that of group B(P<0.05),and the sperm PR,PR+NP level of group F was higher than that of group B(P<0.05).Johnson’s score was found in the HE staining section,which was lower in group B than in group A(P<0.05);Group C,group D,group E,and group F increased compared with group B(P<0.05).Experimental study 2:Immunofluorescence:Bcl-2 was evenly distributed in the seminiferous tubule space in group A.compared with group A,the staining amount of Bcl-2 in group B decreased after microwave irradiation,and the average fluorescence intensity of Bcl-2 in group B decreased(P<0.05).The main expression regions moved from the seminiferous tubule space to the seminiferous tubule;After the intervention,the expression of Bcl-2 increased.The average fluorescence intensity of Bcl-2 in group D,E,and F were higher than that in group B(P<0.05);In group A,only a small amount of Bax staining was scattered.After microwave irradiation,group B was significantly higher than group A,and the main expression regions were concentrated in seminiferous tubules.After the intervention,the staining of Bax in group C,group D,group E,and group F was lower than that in group B;Caspase-3 staining was only scattered in group A,and significantly increased in group B.After the intervention,caspase-3 staining decreased in group C,group D,group E and group F compared with group B(P<0.05).Cytokine:compared with group A,The contents of TNF-α IL-6 and IL-12 in testicular tissue、in group B were higher than those in group A(P<0.05);TNF-α and IL-12,in group C,group D,group E,group F decreased compared with group B(P<0.05),and in IL-6,group E,group F decreased compared with group B(P<0.05).There was no significant difference between group C and group B(P>0.05).Oxidative stress:after the intervention,compared with group A,GSH_PX and MDA in testes of mice in group B increased(P<0.05);Compared with group B,GSH_PX and MDA of mice in groups D,E,and F decreased(P<0.05),CAT decreased in Group E(P<0.05);There was no significant difference among other groups.Conclusions:Clinical research:Professor Bi Huan-zhou has many prescriptions for the treatment of male infertility,such as tonifying the kidney and spleen,removing blood stasis,and dredging collaterals,which are guided by the "theory of successive days" and the "theory of treating both symptoms and signs" and the drug characteristics of Tonifying the spleen and kidney,eliminating pathogenic factors and strengthening health,which can provide ideas for the clinical treatment and scientific research of male infertility in the future.Experimental study 1:microwave radiation can reduce the sperm motility of mice,and the sperm motility can be increased to varying degrees after the intervention.From the HE staining of testicular tissue,it can be seen that microwave radiation may lead to the destruction of seminiferous tubule structure,the disappearance of Sertoli cells,and spermatogenesis dysfunction.Hailong Huangjing powder has a protective effect on testicular seminiferous tubules of mice with oligoasthenospermia induced by microwave radiation,and sperm motility is improved.Acupuncture and moxibustion show a similar effect.Experimental study 2:acupuncture "Sanyinjiao(SP6)" has a protective effect on sperm motility of mice with oligoasthenospermia induced by microwave radiation.One of its mechanisms may be that Hailong Huangjing powder can reduce the level of TNF-α、IL-6,IL-12 in testicular spermatogenesis microenvironment,and other inflammatory cytokines,thereby reducing GSH_Px,MDA mediated oxidative stress further regulates Bcl-2,Bax,caspase-3 and other apoptotic signaling pathways. |
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