Effects Of Icariin On NO Production And NOS Activity Of Rabbit Clitoral Corporal Smooth Muscle Cells

Objectives: To realize the biological characteristics of rabbit clitoral smooth muscle cells (RCSMC) and the effects of Icariin on RCSMC, the effects of Icariin on nitric oxide (NO) production and nitric oxide synthase (NOS) activity of RCSMC were investigated.Methods: In vitro the RCSMC were cultured by enzymatically dispersed method and cell culture technology, morphological observation, cell counting, adhesion rate evaluations, cell growth curve measurement, immunocytochemistry staining were applied to study the biological features of RCSMC. The effects of icarrin on NO production and NOS activity in RCSMC were measured by nitrate reductase test and NOS kit. The cell viability and proliferation treated with icariin evaluated by MTT [3(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium-bromide].Results: Cultured RCSMC exhibited typical morphologic characteristics with spindle shaped and parallel along their longitudinal axis, showing obvious orientation. The attachment and the proliferation of RCSMC in vitro were rapid. Cultured RCSMC could grow and maintain their steady characteristics provided appropriate passage rate and culture condition. NO production and NOS activity in RCSMC treated by icariin were increased significantly in a dose dependent manner (p<0.01), therefore, these responses were significantly inhibited by NOS inhibitor LNNA (p<0.01). The MTT results showed that icariin could increase the RCSMC growth rate in a certain concentration range.Conclusions: The New Zealand White rabbit RCSMC culture model will allow further investigation of the cellular and molecular physiologic mechanisms of female clitoral physiology, receptor-mediated signal transduction mechanisms involved in clitoral smooth muscle contractility, pharmacological interactions. Icariin may increase NO production by activating NOS in RCSMC, and may be effective for clitoral corporal smooth muscle relaxation inducing clitoris tumescence during sexual stimulation. Icariin may also increase RCSMC growth rate in low concentration. The further study is needed.