Shugan Bushen Yulin Decoction(疏肝补肾毓麟汤)Can Improve Sperm Quality of Liver Depression and Kidney Deficiency Type of Oligasthenospermia Mice by Regulating Keap1/Nrf2/GPX4 Pathway to Inhibit Ferroptosis of Testicular Cells
Objective To investigate the effects and possible mechanism of Shugan Bushen Yulin Decoction(疏肝补肾毓麟汤)in improving spermatogenesis and sperm quality of liver depression and kidney deficiency type of oligoasthenospermia mice.Methods Sixty SPF male KM mice were randomly divided into blank group,model group,Shugan Bushan Yulin Decoction high-dose,medium-dose and low-dose groups and statin-1(Ferrostatin 1,Fer-1)group(Fer-1 is ferroptosis inhibitor)by ran-dom number method.Each group consisted of 10 animals.The other groups were given 1.25 g·kg-1 adenine intragastrium(ig)+chronic binding stimulation to construct oligasthenospermia mouse model with liver depression and kidney deficiency.Af-ter drug intervention,the body mass,testicle mass and epididymal mass of mice were measured with precision balance to calculate testicle coefficient and epididymal coefficient.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of testis and score the spermatogenic function of testis.Automatic sperm analyzer was used to detect the sperm density and motility.Sperm maturity was observed by aniline blue staining.Iron deposition in testicular tissue was observed by Bruce blue staining.The expressions of glutathione peroxidase 4(GPX4)and solute vector family 7 member 11(SLC7A11)in testicular tissue were determined by immunohistochemical(IHC).The activities of superoxide Dismutase(SOD),malondialdehyde(MDA)and cata-lase(CAT)in testicular tissue were detected by biochemical method.The expressions of Kelch-like ECH associated protein 1(Keap1),nuclear factor 2 associated factor 2(Nrf2),GPX4 and SLC7A11 in testicular tissue were detected by Western blotting(WB).Results Compared with those of the blank group,the testis coefficient,epididymis coefficient,testis spermatogenic func-tion,sperm density,motility rate and sperm maturity in the model group were significantly decreased(P＜0.05,P＜0.01).The above indexes were significantly improved after high and medium dose and Fer-1 intervention with Shugan Bushan Yulin Decoc-tion(P＜0.05,P＜0.01),and the iron deposition decreased significantly.The protein contents of Keap1 in testicular tissue were significantly decreased,while the protein contents of Nrf2,SLC7A11 and GPX4 were significantly increased(P＜0.05,P＜0.01).Conclusion Shugan Bushen Yulin Decoction can significantly improve liver depression and kidney deficiency type of oli-gasthenospermia,and its mechanism may be related to regulating Keap1/Nrf2/GPX4 signaling pathway and inhibiting ferroptosis of testicular cells.
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