Effect of Baihe Yuzi Formula on Zinc Homeostasis,CFTR and Keap1/Nrf2/HO-1 Signaling Pathway in Oligoasthenospermia Rats
Objective To observe the effects and mechanism of Baihe Yuzi Formula(BYF)on semen parameters and testicular tissue structure.Methods Of the 35 male SD rats,7 rats were randomly selected as the normal group,and the remaining 28 rats were randomly divided into model group,BYF low-dose group,BYF high-dose group and leucovorin group after OAS modeling with tretinoin,7 in each group.The BYF low and high dose groups were given 6.3 and 12.6 g·kg-1·d-1 BYF by gavage,the leucovorin group was given leucovorin 100 mg·kg-1·d-1 by gavage,and the normal and model groups were given an equal amount of saline by gavage.Semen parameters was detected;HE staining was used to observe testicular tissue morphology;blood and testicular tissue zinc content were detected;zinc ion probe was used to detect testicular free zinc ion content;serum testosterone levels and testicular tissue alkaline phosphatase(ALP),lactate dehydrogenase(LDH),alcohol dehydrogenase(ADH),superoxide dismutase(SOD),malondialdehyde(MDA),and glutathione peroxidase(GSH-Px)levels were measured by ELISA;Western Blot was used to detect metal regulatory transcription factor 1(MTF1),zinc transporter(ZNT)4,ZnT8,zinc-iron transporter(Zip)8,Zip12,Kelch-like ECH-associated protein 1(Keap1),NF-E2-related factor 2(Nrf2),heme oxygenase 1(HO-1),histone deacetylase 2(HDAC2),cystic fibrosis transmembrane conductance regulator(CFTR),aquaporin(AQP)3,and AQP9 protein expression.Results Compared with the normal group,the model group had disturbed testicular structure,a few spermatocytes were seen,and free zinc levels were reduced;sperm count and viability,serum and testicular tissue zinc content,serum testosterone level,testicular tissue ADH,ALP,LDH,SOD,GSH-Px content,ZnT4,ZnT8,Zip8,Zip12,MTF1,Keap1,Nrf2,HO-1,HDAC2,CFTR,and AQP9 protein expression decreased in the model group,testicular tissue MDA content,AQP3 protein expression increased(P＜0.05).Compared with the model group,testicular tissues of the intervention group showed different degrees of recovery,increased sperm cell number,and elevated free zinc ion levels;sperm count and viability,testicular tissue zinc content,testicular tissue ALP,LDH,and SOD content,testicular tissue protein expression of ZnT4,ZnT8,Zip8,Zip12,NRF2,HO-1,and HDAC2 increased in the intervention group,and testicular tissue MDA content decreased(P＜0.05).Serum zinc content,serum testosterone level,testicular tissue ADH content,MTF1,KEAP1,CFTR,and AQP9 expression increased,testicular tissue AQP3 expression decreased in the BYF high-dose group(P＜0.05);while serum zinc content,testicular tissue ADH,GSH-Px content,and KEAP1 expression increased,testicular tissue AQP3 expression decreased in the leucovorin group(P＜0.05).Conclusion BYF could effectively improve semen parameters and testicular tissue structure in OAS rats,and its mechanism may be related to regulating zinc homeostasis,CFTR/AQP and Keap1/Nrf2/HO-1 signalling pathways,improving oxidative stress,and increasing serum testosterone.
oligozoospermiaBaihe Yuzi Formulazinc homeostasisoxidative stresscystic fibrosis transmembrane conductance regulatorKelch-like ECH-associated protein 1NF-E2-related factor 2heme oxygenase 1Chinese herbalcompound
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