ABSTRACT
Objective This study was to investigate the ameliorative effects of mangiferin on prostatic fibrosis in benign prostatic hyperplasia(BPH)and the mechanism of action of regulating microRNA(miRNA)-483-3p.Methods The male mice were randomly divided into five groups:normal control group,BPH model control group,finasteride group,mangiferin group,and mangiferin+miRNA-483-3p antagonist group.The mice model of BPH was induced by castration and subcutaneous injection of tes-tosterone propionate.After 30 days,the prostatic collagen deposition was observed by masson and sirius red stain,and the level of hydroxyproline was detected.Prostatic mRNA levels of transforming grouth factor-β1(TGF-β1),mitogen-activated protein kinase 2(MK2),and mitogen-activated protein kinase kinase 6(MKK6),as well as the level of miRNA-483-3p,were detected by quantita-tive real-time PCR.Prostatic protein levels of TGF-β1,MK2,phosphorylated MK2(p-MK2),MKK6,and p-MKK6 were detected by western blotting.Finally,the binding effect of miRNA-483-3p on MK2 was evaluated by luciferase assay.Results Compared to normal control group,the prostatic collagen deposition,mRNA levels of TGF-β1,MK2,and MKK6,as well as protein levels of TGF-β1,p-MK2,and p-MKK6 were significantly increased(P＜0.01),while the miRNA-483-3p level was significantly decreased in BPH model control group(P＜0.01).Compared with BPH model control group,the mangiferin group was able to up-regulate the level of miRNA-483-3p,reduce the mRNA levels of TGF-β1,MK2,and MKK6,as well as the protein levels of TGF-β1,p-MK2,and p-MKK6,and alleviate prostatic collagen deposition.When compared to the mangiferin group,mangiferin+miRNA-483-3p antagomir significantly decreased the miRNA-483-3p level,increased the prostatic collagen deposition,mRNA levels of TGF-β1,MK2,and MKK6,as well as protein levels of TGF-β1,p-MK2,and p-MKK6(P＜0.01).Luciferase assay showed that miRNA-483-3p could tar-get binding with MK2.Conclusion Mangiferin can attenuate prostatic fibrosis by regulating miRNA-483-3p and inhibiting MK2.
ABSTRACT
OBJECTIVE:To investigate the effect of Cyslosorus acuminatus flavonone glycoside (CAF) on kidney epitheli-al-mesenchymal transition(EMT)in rats with diabetic kidney disease(DKD). METHODS:Rats were randomly divided into nor-mal group(normal saline),model group(normal saline),positive group [rosiglitazone,0.4 mg/(kg·d)],CAF high-dose and low-dose groups [12.5,25 mg/(kg·d)],10 in each group. Except for normal group,other groups were intraperitoneally injected strepto-zotocin(60 mg/kg)+high fat diet to induce DKD,and intragastrically administrated related medicines in 13-16 weeks. After the ex-perimental period,fasting blood glucose level and serum creatinine(Scr),blood urea nitrogen(BUN)contents of rats were detect-ed,collagen deposition and basement membrane thickening in kidney tissue were observed. Immunohistochemistry was used to de-tect α-smooth muscle actin(α-SMA),fibronectin,epithelial cadherin(E-cadherin)expressions in kidney tissue,and Western blot was used to determine the glycogen synthase kinase 3β(GSK-3β),phosphorylated GSK-3β(p-GSK-3β),β-catenin expressions in kidney tissue. RESULTS:Compared with normal group,fasting blood glucose level,Scr and BUN contents in model group were significantly increased (P<0.01);kidney tissue showed obvious collagen deposition and basement membrane thickening;theα-SMA,fibronectin,β-catenin expression levels and GSK-3β phosphorylation degree in kidney tissue were significantly increased (P<0.01),while E-cadherin expression levels was significantly decreased(P<0.01). Compared with model group,fasting blood glucose level,Scr and BUN contents in each administration group were significantly reduced(P<0.05 or P<0.01);collagen depo-sition and basement membrane thickening in kidney tissue were significantly improved;the α-SMA,fibronectin,and β-catenin ex-pression levels and GSK-3β phosphorylation degree in kidney tissue were significantly decreased (P<0.05 or P<0.01),while E-cadherin expression levels in positive group and CAF high-dose group were significantly increased(P<0.01). CONCLUSIONS:CAF can inhibit the kidney EMT of rats with DKD,the molecular mechanism may be associated with downregulating β-catenin ex-pression and inhibiting GSK-3βphosphorylation inactivation.