高糖、高胰岛素对血管内皮粘附功能的影响

Effect of high-glucose and high-insulin on the adhesive function of vascular endothelial cells

目的和方法：采用５１Ｃｒ标记血小板与内皮细胞（ＥＣ）粘附，ＥＬＩＳＡ方法检测ＥＣ表面粘附受体－ＶｎＲ（即整合素αｖβ３）改变；Ｆｕｒａ－２／ＡＭ负载测定胞内游离钙等方法，观察高糖、高胰岛素对培养的ＥＣ粘附功能的影响。结果：（１）高糖（３０ｍｍｏｌ／Ｌ）可明显促进ＥＣ与血小板的粘附（ｃｐｍ为３６１００±２１９３ｖｓ２１９６７±１６２６，Ｐ＜００１）。抗ＶｎＲ－β单抗可阻断ＥＣ与血小板粘附，与同刺激物未加单抗组相比有显著差异（ｎｏｎ－Ａｂ３６１００±２１．９３ｖｓＡｂ２７７．６０±１０．４３，ｎ＝６，Ｐ＜００１）。高胰岛素（１５０ｎｍｏｌ／Ｌ）有相同作用（２９０８３±１６８３ｖｓ２１９．６７±１６．２６，Ｐ＜００１，加单抗后ｎｏｎ－Ａｂ２９０００±１６．８３ｖｓＡｂ２２８．１７±１８．６１，Ｐ＜００１）。（２）高糖、高胰岛素均可使ＥＣ的ＶｎＲ表达增加。但过高量（５０ｎｍｏｌ／Ｌ葡萄糖、２５０ｎｍｏｌ／Ｌ胰岛素）使ＶｎＲ减少，伴有ＥＣ损伤。正常量胰岛素（５０ｍｍｏｌ／Ｌ）抑制ＶｎＲ表达，但ＥＣ形态正常。（３）高糖使胞内Ｃａ２＋浓度（ｎｍｏｌ／Ｌ）明显增加（８９６３３±２５１５ｖｓ２１４?

AIM and METHOD:This study was performed on culture endothelial cells (EC) to observe the effect of high-glucose(HG) and high-insulin(HI) on the adhesion of ECs to platelets(PLS) and its mechanism cells RESULTS:HG(30mmol/L) enhanced obviously the adhesion of EC to PLs, the cpm of HG group were 361 00±21.93 and control group were 219.67±16.26 ( P <0 01). Anti-β 3 monoclonal antibody could partly block the effect of this adhesion. There was a remarkable difference between two groups, the cpm were 219.67±16.26 in HG+nonMcAb group and 277 60±10 43 in HG+McAb group ( n =6, P <0 01). The effect of HI (150nmol/L)was similar to HI (290.83±16.83 in HI+McAb group 219.63±16.26 in control and 228.17±18.61 in HG+McAb group,( n =6 P <0 01). HG and HI could increase the expression of αvβ 3,but over-dosage of HG and HI(HG>40mmol/L,HI>200nmol/L) reduced denisities of αvβ 3 with damage of EC. The normal concentration (50nmol/L) of insulin supressed expression of αvβ 3 mildly with normal morphology of EC. HG could increase [Ca 2+ ]i remarkablly (893.33±15.15)nmol/L in HG group vs . (214 00±19.59)nmol/L in control ( n =5, P <0 001),HI induced [Ca 2+ ]i increase also. CONCLUSION: HG and HI could activate EC respectively, enhanced the adhesion of PL to EC, increased[Ca 2+ ]i and αvβ 3 was involved in this mechanism as a very important medium. However, over-concentration of HG reduced expression of αvβ 3 and damaged EC. .