绞股蓝总皂苷对内毒素、氧自由基介导血管内皮细胞表达c-sis基因和调节平滑肌细胞增殖的影响

Effects of gypenosides on c sis expression of bovine aortic endothelial cells and proliferation of vascular smooth muscle cells induced by lipopolysaccharide and active oxygen species

目的 研究绞股蓝总皂苷（ Ｇｙｐ） 对血管平滑肌细胞增殖的调节与内皮细胞ｃｓｉｓ 基因表达和一氧化氮（ Ｎ Ｏ） 合成／ 释放的关系。方法 采用原位杂交法检测内皮细胞ｃｓｉｓｍ Ｒ Ｎ Ａ 表达，用 Ｇｒｉｅｓｓ 法测定内皮细胞 Ｎ Ｏ 释放量，用 Ｍ Ｔ Ｔ快速比色法检测平滑肌细胞（ Ｓ Ｍ Ｃ） 增殖。结果 内毒素（ Ｌ Ｐ Ｓ）１６ ｍｇ· Ｌ－ １ 促进牛主动脉内皮细胞ｃｓｉｓ 基因表达，明显降低内皮细胞培养液中一氧化氮含量，刺激 Ｓ Ｍ Ｃ 增殖；黄嘌呤－ 黄嘌呤氧化酶（ Ｘ Ｘ Ｏ） 系统产生的氧自由基（ Ｏ Ｆ Ｒ） 具有与 Ｌ Ｐ Ｓ 相似的作用。绞股蓝总皂苷对 Ｌ Ｐ Ｓ诱导的内皮细胞ｃｓｉｓ 基因表达具有明显抑制作用，并能对抗 Ｏ Ｆ Ｒ 诱导的 Ｅ Ｃ 损伤，保护内皮细胞释放 Ｎ Ｏ 的能力，抑制 Ｌ Ｐ Ｓ 和 Ｏ Ｆ Ｒ通过 Ｅ Ｃ 介导的 Ｓ Ｍ Ｃ 增殖。 Ｇｙｐ 的效应可被 Ｎ Ｏ 合成酶抑制剂硝基左旋精氨酸部分取消。结论 绞股蓝总皂苷通过抑制内皮细胞ｃｓｉｓ 基因表达，促进内皮细胞合成（ 释放） Ｎ

AIM To investigate the effects of gypenosides(Gyp) on c sis expression of endothelial cells (EC) and proliferation of vascular smooth muscle cells (VSMC)induced by lipopolysaccharide (LPS) and oxygen free radical(OFR) METHODS Cultured bovine aortic endothelial cells were incubated with Gyp (10～ 20 mg·L -1 )and LPS (16 mg·L -1 ) or xanthine(100 μmol·L -1 ) plus xanthine oxidase (100 U·L -1 ) C sis expression was determined by in situ hybridigation Nitrite, the matabolic product of NO, was measured according to a modified Griess method,and the proliferation of endothelial cells and VSMC was estimated by MTT assay. RESULTS We observed that LPS promoted c sis gene expression, and inhibited NO release, and that X XO inhibited EC proliferation and NO release Both LPS and X XO stimulated SMC proliferation by inducing growth factors release from EC Gyp significantly decreased c sis gene expression,increased NO synthesis and proliferation of EC Furthermore, Gyp indirectly inhibited SMC proliferation by blocking the effects of LPS and X XO. The effects of Gyp were abolished by NO synthase inhibitor N G L Arg CONCLUSION Gyp inhibited X XO and LPS induced proliferation of VSMC by attenuating c sis gene expression and promoting NO synthesis/release of EC.