Intervention Effect Of Ursolic Acid On Adriamycin-induced Nephropathy In Mice

Objective: Study of the effect of ursolic acid on adriamycin-induced nephropathy in mice, toexplore its possible mechanism, and to provide the experimental basis for preventing ordelaying the development of renal fibrosis, while seeking new drugs for the prevention andtreatment of renal fibrosis.Methods: Forty male Balb/c mice of18-22g, were randomly divided into Normal group(n=10), ADN group (n=10), LU+ADN group (n=10), HU+ADN group (n=10). At day4,ADN group, LU+ADN group and HU+ADN group was treated with a single tail veininjection of ADR at10mg/kg. The Normal group received a single tail vein injection ofsaline only. From day1to18，LU+ADN group was given UA25mg/(Kg d)(dissolved by14%DMSO) via intraperitoneal injection once a day. HU+ADN group was given UA50mg/(Kg d)(dissolved by14%DMSO). Normal group and ADN group was given the sameamount of vehicle (14%DMSO) by intraperitoneal injection. Seven days after ADRtreatment, twenty four-hour urine was collected with using metabolic cages. Fourteen daysafter ADR treatment, all mice were killed and serum was collected biochemical parameterstests，kidneys were immediately harvested histological analyses. HE staining was used toobverse the tubulointerstitial lesions, and Immunohistochemical staining detected theexpression of TGF-β1in tubulointerstitium.Results:1. General condition and body weight：After ADR injection, the mice were less foodand water intake less and their activity decrease obviously. The body weight of Normal groupincrease over time. The body weight of other group decrease after ADR injection.2.24-hoururinary protein excretion：At day7after ADR injection, albuminuria was most evident inADN group as compared with Normal group (P<0.05), and was attenuated in HU+ADNgroup (P<0.05).3. Renal function：Serum creatinine and BUN were detemnined to reflectrenal function. The serum creatinine and BUN were no statistical difference among allgroups.4. Renal oxidative stress: Serum MDA were detemnined to reflect renal oxidativestress. ADR mice had elevated plasma MDA (P<0.01), both of which were no significantly attenuated in UA treatment group.5. HE staining: Renal tissues were not significant changesin Normal group. The renal tubulointerstitial lesions of ADN group mice were obvious, withswelling of tubular epithelial cells, tubular dilation obviously, the tubulointerstitial lesionsdeveloped the fibrosis. The tubulointerstitial lesions of HU+ADN group mice were relativelyrelieved.6.The semi-quantitative analysis of Immunohistochemical staining:①Comparedwith Normal group, the difference of TGF-β1expression in ADN group renal tubularepithelial cells were statistically significant(P<0.05).②Compared with ADN group, thedifference of TGF-β1expression in LU+ADN group were no statistical difference; thedifference of TGF-β1expression in HU+ADN group were statistically significant(P<0.05).Conclusion:1.This experiment confirmed that higher doses of UA (50mg/Kg) can reduceurinary protein in adriamycin-induced nephropathy in mice.2. Initially confirmed the higherdose of UA (50mg/Kg) can inhibit the TGF-β1protein expression in tubulointerstitial ofadriamycin-induced nephropathy in mice, inhibit or delay the progress of glomerulosclerosisand tubule interstitial fibrosis, play a renoprotective role in adriamycin-induced nephropathyin mice.3. The present study suggest the therapeutic potential of UA for management ofhuman glomerular disease.