Wuzi Yanzong Pill improves spermatogenic function in obese rats by regulating mitochondrial fusion protein 2 and reducing endoplasmic reticulum stress in testis
Objective To observe the effect of Wuzi Yanzong Pill(WYP)on spermatogenic function in obese rats induced by high fat diet,and to explore the possible mechanism from ERS in testis.Methods Totally 30 male SD rats were divided into control group,model group,low,middle and high dose(0.5,1.0,and 2.0 g·kg-1)groups of WYP randomly.The control group was fed with normal diet,the rest were fed with high-fat diet.After 16 weeks,the corresponding doses of drugs in each group were gavaged for four weeks continuously.the rats were dissected,and sperms in comepididymis were collected for the function analysis.Serum ELISA was used to analyze the changes of sex hormone levels.The pathological changes of testis were observed by hematoxylin-eosin staining.Transmission electron microscopy(TEM)was used to observe the structural changes of the mitochondria and endoplasmic reticulum in sertoli cells.The localization and expression of Mfn2 and GRP78 in testis were detected by immunofluorescence staining.The expression of Mfn2 and PERK pathway related proteins in testis were detected by Western blotting.Results Compared with control group,the model group rats showed a significant increase in body weight(P＜0.01),a marked increase in serum estradiol level(P＜0.01),and a marked decrease in serum testosterone level(P＜0.01);the arrangement of spermatogenic cells in the testes was sparse,and the cells were degenerated;the sperm motility was significantly lower(P＜0.05),and the sperm malformation rate was markedly increased(P＜0.01);the rough endoplasmic reticulum and mitochondria in the Leydig cells were swollen and hollow,some of the network was broken,the expression of Mfn2 was markedly reduced,and the GRP78 fluorescence expression was markedly increased;the protein levels of PERK,activated transcription factor-4(ATF4),and CHOP were markedly increased(P＜0.01).Compared with the model group,the high-dose group of Wuzi Yanzong Pill showed a significant reduction in body weight(P＜0.05);the serum estradiol level was markedly lower in the middle and high-dose groups(P＜0.05,0.01),and the serum testosterone level was markedly higher(P＜0.05,0.01);the sperm motility was significantly improved in the high-dose group(P＜0.05),and the sperm malformation rate was markedly reduced(P＜0.05);the pathological changes in the testes were alleviated,the arrangement was denser,and the rough endoplasmic reticulum and mitochondria in the Leydig cells were improved;the fluorescence expression of Mfn2 gradually increased,and the relative protein expression was markedly increased(P＜0.05,0.01),while the GRP78 fluorescence expression gradually decreased;the ATF4 protein level was markedly reduced in the low-dose group(P＜0.01),the PERK and ATF4 protein levels were markedly reduced in the middle-dose group(P＜0.05,0.01),and the PERK,ATF4,and CHOP protein levels were all markedly.Conclusion Wuzi Yanzong Pill can ameliorate spermatogenesis disorder induced by high fat diet in rats,which may be related to the regulation the expression of Mfn2 and alleviate the endoplasmic reticulum stress in sertoli cells.
Wuzi Yanzong Pillobesityspermatogenesistestissertoli cellsmitochondrial fusion protein 2eendoplasmic reticulum stress
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