Study on Mechanism of Shenqu Zhuyu Decoction in Improving Varicocele Rats Spermatogenesis Based on PINK1/Parkin Mediated Mitophagy and Apoptosis
Objective To observe the effect of the Shenqu Zhuyu Decoction(SZD)on varicocele(VC)rats mitophagy and apoptosis in left testis based on PTEN-induced putative kinase 1(PINK1)/E3 ubiquitin protein ligase(Parkin)signaling pathway,and to explore the mechanism.Methods Totally 36 male SD rats were randomly divided into sham operation group,model group and SZD group,12 rats in each group.VC rats model was established by partial ligation of the left renal vein in model group and SZD group.At 48th day after successful molding,the rats in SZD group were treated with SZD(0.7 g/100 g)by gavage,and rats in sham operation group and model group were given the equal volume of normal saline by gavage,twice a day for 60 days.The left testicle of the rats in each group were taken.HE staining was used to observe the morphology of testicles.Transmission electron microscopy was used to observe the ultrastructure of mitochondria and mitophagosomes in testicular tissue.Western Blot was used to detect protein expression of PINK1,Parkin,microtubule associated protein 1 light chain 3 beta(LC3B)Ⅱ/Ⅰ,sequestosome1(p62),B-cell lymghoma(Bcl-2),Bcl-2 associatal X protein(Bax),Cleaved Cysteine-containing aspartate-specific protease-3(Cleaved Caspase-3)in testicular tissue.The protein expressions of PINK1 and Parkin in testicular tissue were aslo detected by immunohistochemistry.Results Compared with the sham operation group,pathological changes occurred in the testis tissue of the model group,the mitochondria were swollen with irregular shapes,accompanied by vacuolization and mitochondrial crest rupture,increasing mitochondrial autophagosome formation was observed under transmission electron microscope,the expression of autophagy-related proteins PINK1,Parkin,LC3BⅡ/Ⅰ increased(P＜0.01),expression of p62 protein decreased(P＜0.01),the expression of apoptosis-related proteins Bax,Cleaved Caspase-3 increased(P＜0.01,P＜0.05),while expression of Bcl-2 decreased(P＜0.05).Compared with model group,pathological changes of testicular tissue and mitochondrial damage were recovered in SZD group,the formation of mitophagosomes decreased,the expression of autophagy-related proteins PINK1,Parkin,LC3BⅡ/Ⅰ decreased(P＜0.01,P＜0.05),while expression of p62 increased(P＜0.01),the expression of apoptosis-related proteinsc Bax,Cleaved Caspase-3 decreased(P＜0.05),while expression of Bcl-2 increased(P＜0.05).Conclusion SZD could improve the spermatogenic function of VC rats by regulating PINK1/Parkin-mediated mitophagy and reducing apoptosis of spermatogenic cells.
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