The Therapeutic Effect Of The Roots Of Rhododendron Molle And Its Component-Rhodojaponin ? In Mice With Adriamycin Nephropathy And Its Mechanism

Rhododendron molle G.Don(RM)is a natural Chinese herbal medicine,which was traditionally used to treat rheumatoid arthritis and bruises.In the past 30 years,RM has been used in clinic to treat chronic nephritis,and proved to be effective.Focal segmental glomerulosclerosis(FSGS)is a clinicopathological syndrome with unknown causes and continuous progression.The characteristic clinical manifestations of FSGS are heavy proteinuria,hypoalbuminemia,hyperlipidemia,insensitive to hormone therapy,and progression to end-stage renal disease(ESRD).The main pathological features are focal and segmental glomerular sclerosis and tubulointerstitial fibrosis.In recent years,with the development of plant chemical separation technology,many bioactive compounds were separated from RM.In this study,we tested root ethanol extract of RM and its important compound rhodojaponin II(R-II)in mice model of adriamycin nephropathy(ADRN),which could simulate human FSGS,to identify effective compound of RM for the therapy of kidney diseases and to explore its mechanism.Part ? Therapeutic effect of the roots extract of Rhododendron molle in adriamycin nephropathy miceObjective: To test the therapeutic effect of the roots extract of Rhododendron molle in ADRN mice.Methods:(1)ADRN was established by single intravenous injection of adriamycin(ADR)15 mg/kg to 8 weeks old male Balb/c mice.(2)ADRN mice were given intragastrical administration of the following medications for 5 weeks: low dose of the roots extract of RM(5 mg/kg),high dose of the roots extract of RM(60 mg/kg),and positive control captopril(30 mg/kg).(3)Gomerular injury was evaluated by PAS staining.(4)Kidney fibrosis was evaluated by Sirius red staining and Masson staining.(5)The expressions of podocyte marker(nephrin and podocin)were detected with immunohistochemical staining.(6)The expression of fibronectin was detected with immunohistochemical staining.Results:(1)Low dose of the roots extract of RM could significantly attenuate proteinuria and hypoalbuminemia in ADRN mice.(2)Low dose of the roots extract of RM could significantly attnuate glomerular sclerosis and improve nephrin and podocin expressions in ADRN mice.(3)Low dose of the roots extract of RM could significantly decrease positive area of Sirius red staining and Masson staining,and fibronectin expression in ADRN mice.Conclusion: Low dose of the roots extract of RM could effectively reduce proteinuria and glomerulus injury,and delay kidney fibrosis in ADRN mice.Part ? Therapeutic effect of rhodojaponin II in adriamycin nephropathy mice and its mechanismObjective: To test the therapeutic effect of rhodojaponin II in ADRN mice,and to explore its mechanism.Methods:(1)ADRN was established by single intravenous injection of ADR 15 mg/kg.(2)8 weeks old male Balb/c mice were randomly divided into 4 groups:Sham control group(Sham),Sham control+R-II 0.04 mg/kg group(Sham+R-II 0.04),adriamycin nephropathy group(ADR),and adriamycin nephropathy+R-II 0.04mg/kg group(ADR+R-II 0.04).R-II was given intragastrically for 5 weeks.(3)Glomerular injury was evaluated by PAS staining.(4)Kidney fibrosis was evaluated by Sirius red staining and Masson staining.(5)The expressions of nephrin and podocin were detected with immunohistochemical staining and Western blot analysis.(6)The expression of fibronectin was detected with immunohistochemical staining and Western blot analysis.(7)Interstitial infiltrated inflammatory cells(CD4+ T cells,CD8+ T cells,and CD68+ macrophages)were detected with immunohistochemical staining.(8)The expression of NF-?B p-p65 was detected by Western blot analysis.(9)The expressions of TGF-?1/Smad pathway associated key proteins,such as TGF-?1,p-smad3,Smad3,and Smad7,were detected by Western blot analysis.Results:(1)R-II could significantly attenuate proteinuria and hypoalbuminemia in ADRN mice.(2)R-II could significantly attenuate glomerular sclerosis and improve nephrin and podocin expressions in ADRN mice.(3)R-II could significantly reduce positive area of Sirius red staining and Masson staining,and fibronectin expression in ADRN mice.(4)R-II could significantly attenuate interstitial infiltration of CD4+ T cells,CD8+ T cells,and CD68+ macrophages in ADRN mice.(5)R-II could significantly reduce the phosphorylation of NF-?B p65 in ADRN mice.(6)R-II could significantly downregulate the expressions of TGF-?1 and p-Smad3 in ADRN mice.Conclusions:(1)Rhodojaponin II could significantly reduce proteinuria and kidney injury in ADRN mice,and it is the effective compound of the roots of Rhododendron molle for the treatment of ADRN.(2)The mechanisms are related to its anti-inflammatory effect and regulation of TGF-?1/Smad signaling pathway.