茶色素对培养猪离体主动脉内皮细胞功能影响的实验研究

The Effects of Tea Pigment on Functions of Porcine Aorta Endothelial Cells in vitro

目的利用体外培养的猪主动脉血管内皮细胞的实验模型，研究茶色素（ＴＰ），左旋精氨酶（Ｌ－Ａｒｇ），ＮＯ合酶抑制剂（Ｌ－ＮＡＭＥ）对血管内皮细胞增殖及分泌一氧化氮（ＮＯ）、内皮素（ＥＴ）的干预作用。方法制备３～６代猪主动脉血管内皮细胞的单细胞悬液。分为六个组，每组８孔，５×１０４个细胞／孔。ａ．对照组，ｂ．Ｌ－Ａｒｇ（１０μｍｏｌ／Ｌ）组，ｃ．Ｌ－ＮＡＭＥ（１０－２μｍｏｌ／Ｌ）组，ｄ．茶色素分低（１０μｇ／ｍｌ）、中（５０μｇ／ｍｌ）、高（１００μｇ／ｍｌ）剂量三个组，培养１，３，５天后分别进行细胞计数，培养４８小时后测培养液中的ＮＯ及ＥＴ含量。结果（１）细胞生长试验：Ｌ－Ａｒｇ组与对照组相比细胞数明显减少，而Ｌ－ＮＡＭＥ与对照组相比，细胞数明显增加。茶色素低、中、高剂量三个组分别与对照组相比，细胞数均明显减少，其抑制作用与剂量相关；（２）ＰＡＥＣ分泌ＮＯ及ＥＴ含量：Ｌ－Ａｒｇ组与对照组相比，ＥＴ降低，ＮＯ升高；而Ｌ－ＮＡＭＥ与对照组相比，ＥＴ增高，ＮＯ降低。茶色素低、中、高剂量三个组与对照组相比，ＮＯ升高，ＥＴ降低，并与剂量相关。结论茶色素抑制离体培养的ＰＡＥＣ增殖，抑制ＥＴ分泌，促进ＮＯ释放，且与剂量相关?

Objective\ We investigated the effect of tea pigment on the release of nitric oxide(NO) and endothelin(EN) in culture endothelial cell from porcine aorta. The effect of L arginine (L Arg) and N G nitro arginine methylester(L NAME) on the NO,ET were studied and compared with the effect of tea pigment.\ Methods\ PAEC was isolated by collagenase and pancreatin digestion. The 3th～6th generation of cultured PAEC were used with the cell count of 5×10 6/ml in each core.The experiment consisted of 6 groups:(A)control group;(B)L Arg(10 μmol/L) group;(C)L NAME(10 -2 μmol/L) group;(D)three kind of dosages tea pigment:Low(10 μg/ml),middle(50 μg/ml), and high(100 μg/ml).\ Results\ L Arg inhibited the proliferation of PAEC. L NAME stimulated proliferation of PAEC, compared with the control group. Tea pigment inhibited the growth of PAEC in a dosage dependent manner. L Arg(10 μmol/L) reduced ET and enhanced NO release.L NAME reduced NO and enhanced ET release,compared with the control group.The tea pigments were dose dependently reduced the release of ET,and increased the release of NO. Conclusions\ L Arg and three dosages tea pigment can inhibit prolifertion of PAEC in vitro and inhibit releasing of ET, and increase the secretion of NO.The effects of L NAME were inverse, compared with that of L Arg.