| OBJECTIVEMale infertility has become a hot issue of social concern.There are some limitations in western medicine treatment,and a considerable part of male infertility can not be effectively solved;Traditional Chinese medicine treatment has unique advantages and exact curative effect,which can make up for the deficiency of Western medicine in treating male infertility;Guangsi Yulin decoction is based on the principle of Tonifying the kidney and promoting sperm,with exact clinical curative effect,which is worthy of further discussion;This study is carried out through a randomized controlled trial step to clarify the clinical effect of Guangsi Yulin decoction,and to explore the mechanism of improving spermatogenic function and the correlation between SCF/c-kit-pi3k-bcl-2 signal pathway through animal experimentsMETHODClinical study:the method of randomized controlled trial was used.The positive control group was treated with Levocarnitine,and the observation group was treated with Levocarnitine combined with Guangsi Yulin decoction.The course of treatment was 90 days The semen quality indexes such as semen quantity,liquefaction degree,sperm concentration,total sperm count,total sperm activity rate,forward movement percentage and the score of TCM symptoms were observed before and after treatment through statistical analysis,the therapeutic effect of Guangsi Yulin Decoction on male infertility with kidney deficiency syndrome was observedExperimental study:Tripterygium wilfordii polyglycoside was used to induce the establishment of spermatogenic disorder rat model.After the success of the model,it was randomly divided into blank group,model group,control group,low dose group,medium dose group and high dose group,12 rats in each group;Blank group and model group were given deionized water,control group was given Levocarnitine oral liquid,low,medium and high dose group were given low,medium and high doses of guangsiyulin Decoction suspension;The rats were given gavage for 4 weeks.The general condition,body weight,organ coefficient of testis and epididymis,semen quality,serum sex hormone,inhibin B,carnitine of epididymis,morphological and pathological changes of testis and ultrastructural changes were observed,and the best therapeutic concentration of guangsiyulin Decoction was discussed.The content of Bax and bcl-2 protein in rat testis was detected by immunohistochemistry;The content of SCF/c-kit-pi3k-bcl-2 signal pathway related protein and mRNA expression were detected by WB and real time PCR;The mechanism of Guangsi Yulin Decoction improving spermatogenesis and its correlation with SCF/c-kit-pi3k-bcl-2 signal pathway were discussedRESULTClinical research:After treatment,the semen quality related indexes in the observation group were significantly better than those in the control group,with significant difference(P<0.05 or P<0.01);The total score of TCM symptoms and different syndrome scores of Syndrome of deficiency of kidney essence,Syndrome of deficiency of kidney-yang,and Syndrome of deficiency of kidney-yin in the observation group were also better than those in the control group,with significant difference(P<0.05 or P<0.01);According to the clinical efficacy standard,the overall efficacy of Guangsi Yulin decoction combined with Levocamitine was significantly better;The difference was statistically significant(P<0.05)No adverse reactions were reported during the whole observation periodExperimental study:After the intervention,the general situation,body weight,organ coefficient,semen quality,inhibin B,carnitine of epididymis and other indexes of rats in each dose group of Guangsi Yulin Decoction were significantly better than those in the model group,and the middle dose group was better than those in the control group;The sex hormone related indexes of Guangsi Yulin Decoction were better than those in the model group,with statistical differences(P<0.05);The middle dose group LH and high dose group T were better than the control group.The morphology and quantity of spermatogenic cells and the number of organelles such as mitochondria in Guangsi Yulin Decoction group were significantly better than those in model group and control group.It was found that the content of Bax in guangsiyulin Decoction group was significantly lower than that in the control group.and the content of Bcl-2 and the ratio of Bcl-2/Bax were significantly higher than that in the control group;It can be seen that the protein content of SCF,c-kit,PI3K,Akt and Bcl-2 in the observation group was higher than that in the control group,and the protein content of bad and Bax were lower than that in the control group.The expression of SCF,c-kit,Akt and Bcl-2 in the observation group was higher than that in the control group,and the expression of bad and Bax mRNA was lower than that in the model group.CONCLUSION1.Guangsi Yulin decoction can effectively improve the semen quality of male infertility patients,reduce the TCM symptom score of kidney deficiency syndrome patients;2.Guangsi Yulin decoction can effectively improve the general state of spermatogenesis disorder rats,improve organ coefficient,semen quality,inhibin B and epididymis carnitine content,regulate hypothalamus pituitary gonadal axis sex hormone,repair the morphology of testicular tissue and ultrastructure;3.The mid-dosage of Guangsi yulin decoction is the best concentration of treatment drug;4.Guangsi Yulin decoction can promote the proliferation of spermatogenic cells and inhibit the apoptosis of spermatogenic cells through SCF/c-kit-PI3K-bcl-2 signaling pathway.In conclusion,Guangsi Yulin decoction can effectively treat male infertility of kidney deficiency type,and its regulation of SCF/c-kit-PI3K-bcl-2 signaling pathway protein and gene is one of the mechanisms. |
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