Effects Of Clerodendranthus Spicatus And Ursolic Acid On TGF-β₁ Expression And The Proliferation Of Cultured Rat Glomerular Mesangial Cells

Objective To investigate the effects of extractions of C.Spicatus and ursolic acid on TGF-β1 expression and the proliferation of cultured rat glomerular mesangial cells,which will be the foundations of Exploitation and Application of C.Spicatus.Methods Primary cultures of glomerular mesangial cells were prepared from month 1-2 SD rats. Mesangial cells were identified by inverted microscope and immunohistochemistry. Ethyl acetate,n-butanol fractions were separated from the ethanol extracts of C.Spicatus. The experimental rats were randomly divided into 8 groups. Rats was drenched the different extracts of C.Spicatus solution at the high- and low- dosages of 10g·kg-1·d-1,2.5g·kg-1·d-1 or ursolic acid solution at the high-, moderate- and low- dosages of 100mg·kg-1·d-1, 50 mg·kg-1·d-1, 25mg·kg-1·d-1 . The drugs-containing serum were separated after 7days. The effects of drugs-containing serum on the proliferation of MC stimulated by LPS were observed by Cell Counting Kit-8(CCK-8) colorimetric assay respectively. The protein release of TGF-β1 in mesangial cells was measured by ELISA and the expression of TGF-β1mRNA was measured by RT-PCR.Results 1. LPS group could obviously stimulate the proliferation of MCs, the protein release of TGF-β1 and the expression of TGF-β1mRNA in MCs. LPS group was more obvious than that of blank group, for there was significant difference(P<0.01).2. The drugs-containing serum of ursolic acid , Ethyl acetate and n-butanol fractions of C.Spicatus extractions could significantly inhibit the proliferation , the protein release of TGF-β1 in mesangial cells induced by LPS (P<0.01;P<0.05); The most remarkable effect was observed by n-butanol fraction of C.Spicatus than that of Ethyl acetate fraction (P<0.01). Some of these effects was dose-dependent (P<0.01; P<0.05). 3. Ursolic acid serum, Ethyl acetate and n-butanol fractions of C.Spicatus extractions serum could significantly inhibit the expression of TGF-β1mRNA in mesangial cells induced by LPS(P<0.01;P<0.05); The effect of n-butanol fraction was superior than that of Ethyl acetate fraction (P<0.01). All these effects was dose-dependent(P<0.01).Conclusion 1. LPS could obviously stimulate the proliferation of MCs, the protein release of TGF-β1 and the expression of TGF-β1mRNA in MCs. 2. The drugs-containing serum of Ethyl acetate and n-butanol fractions of C.Spicatus extractions could significantly inhibit the proliferation,the protein release of TGF-β1 and the expression of TGF-β1mRNA in MCs induced by LPS. The effect of n-butanol fraction was superior than that of Ethyl acetate fraction. 3. The drugs-containing serum of Ursolic acid could significantly inhibit the proliferation,the protein release of TGF-β1 and the expression of TGF-β1mRNA in MCs induced by LPS. Some of these effects was dose-dependent.