Effects Of C.Spicatus And Rosmarinic Acid On The Proliferation And Interleukin-1 β Production Of Cultured Murine Mesangial Cells

Objective To explore the effects of decoction of C.Spicatus and rosmarinic acid on the proliferation and interleukin-1β (IL-1β ) production of cultured murine mesangial cells and to analyze the content of rosmarinic acid in the C.Spicatus decoction.Methods Primary cultures of glomerular mesangial cells were prepared from month 1-2 Spraque-Dawley rats.Mesangial cells were identified by phase contrast microscope,scanning electron microscope and immunohistochemistry.The proliferation of cultured cells stimulated by LPS or sera following incubation with the decoction of C.Spicatus and rosmarinic acid was tested by MTT method. The protein synthesis of IL-1β in mesangial cells induced by LPS was measured by ELISA. Trypanblau test was used to examine the cytotoxicity of C.Spicatus and rosmarinic acid.The content of rosmarinic acid in the decoction of C.Spicatus was measured by HPLC. Results K The C.Spicatus decoction in the range of concentration from 50 u g/ml to 1250)4 g/ml could inhibit the proliferation of mesangial cells induced by sera or LPS(P<0.01); C.Spicatus,m the same range of concentration,also inhibited the protein release of IL-1B (P<0.01).2 Treated by Rosmarinic acid in the range of concentration from 5 u g/ml to 125 u g/ml, the mesangial cells proliferation induced by sera or LPS could be inhibited(P<0.01);in the range of concentration from 1 u g/ml to 125 ug/ml, rosmarinic acid also inhibited the protein release of IL-1β (P<0.01). 3. 250 u g/ml C.Spicatus decoction and 25 u g/ml rosmarinic acid have no cytotoxicity to the mesangial cells(P>0.05),while 1250 u g/ml C.Spicatus decoction and 125 ug/ml rosmarinic acid have obvious cytotoxicity to themesangial cells(P<0.01). 4 The content of rosmarinic acid in the decoction of C.Spicatus was 1.91%.Conclusion The decoction of C.Spicatus in the range of concentration from 50 u g/ml to 250 u g/ml and rosmarinic acid in the range of concentration from 5 u g/ml to 25 u g/ml could inhibit the proliferation of mesangial cells induced by sera or LPS;The C.Spicatus decoction in the range of concentration from 50 u g/ml to 250 u g/ml and rosmarinic acid in the range of concentration from 1 u g/ml to 25 u g/ml could inhibit the protein release of IL-1β induced by LPS; The content of rosmarinic acid in the decoction of C.Spicatus was 1.91%.