葡萄糖和胰岛素对膝下血管平滑肌细胞增殖的相加作用。

The additive effects of glucose and insulin on the proliferation of infragenicular vascular smooth muscle cells.

作者信息

Avena R, Mitchell M E, Neville R F, Sidawy A N

机构信息

Departments of Surgery, Veterans Affairs Medical Center, George Washington and Georgetown University Medical Centers, Washington, DC, USA.

出版信息

J Vasc Surg. 1998 Dec;28(6):1033-8; discussion 1038-9. doi: 10.1016/s0741-5214(98)70029-1.

DOI:10.1016/s0741-5214(98)70029-1

PMID:9845654

Abstract

PURPOSE

Peripheral vascular disease involving the infragenicular arterial tree is common in patients with diabetes mellitus (DM). Accelerated proliferation of vascular smooth muscle cells (VSMCs) plays an important role in the development of atherosclerosis. Insulin and glucose stimulate VSMC proliferation and are elevated in patients with non-insulin-dependent DM. We have previously described the mitogenic effect of insulin on VSMCs in vitro; the effects of insulin and glucose separately and in combination on the proliferation of VSMCs grown in serum-free media were studied.

METHODS

Human infragenicular VSMCs isolated from diabetic patients with end-stage peripheral vascular disease undergoing below-knee amputation were used. Cells from passages 3 to 5 were grown in serum-free media with varying glucose (0.05%, 0.1%, 0.2%, 0.4%, 0.6%, and 0.8%) and insulin (no added insulin, 100 ng/mL, and 1000 ng/mL) concentrations for 6 days.

RESULTS

Insulin stimulated VSMC growth at glucose concentrations more than 0.2% (0.4% glucose with no added insulin resulted in 13,073 +/- 336 cells/mL, 0.4% glucose with 100 ng/mL insulin resulted in 16,536 +/- 1175 cells/mL, 0.4% glucose with 1000 ng/mL insulin resulted in 17,500 +/- 808 cells/mL, 0.6% glucose with no added insulin resulted in 14,167 +/- 1062 cells/mL, 0.6% glucose with 100 ng/mL insulin resulted in 18,984 +/- 1265 cells/mL, 0.6% glucose with 1000 ng/mL insulin resulted in 20,450 +/- 1523 cells/mL, 0.8% glucose with no added insulin resulted in 15, 853 +/- 1650 cells/mL, 0.8% glucose with 1000 ng/mL insulin resulted in 26,302 +/- 1919 cells/mL; P <.05 compared with glucose with no added insulin). Glucose stimulated VSMC proliferation up to a concentration of 0.2% (42% and 117% higher growth at 0.1% and 0.2% glucose, respectively, compared with the baseline, P <.05), regardless of the insulin concentration in the media. The greatest growth (26,302 +/- 1919 cells/mL) occurred in the group with the highest concentration of both insulin (1000 ng/mL) and glucose (0.8% glucose; P <.05).

CONCLUSION

Both insulin and glucose stimulate the growth of diabetic infragenicular VSMCs. The mitogenic effects of insulin and glucose are additive and may contribute to the development of atherosclerosis in patients with DM.

摘要

目的

累及膝下动脉树的外周血管疾病在糖尿病（DM）患者中很常见。血管平滑肌细胞（VSMC）的加速增殖在动脉粥样硬化的发展中起重要作用。胰岛素和葡萄糖刺激VSMC增殖，且在非胰岛素依赖型DM患者中升高。我们之前已经描述了胰岛素在体外对VSMC的促有丝分裂作用；研究了胰岛素和葡萄糖单独及联合对在无血清培养基中生长的VSMC增殖的影响。

方法

使用从接受膝下截肢的终末期外周血管疾病糖尿病患者中分离出的人膝下VSMC。第3至5代细胞在含有不同葡萄糖（0.05%、0.1%、0.2%、0.4%、0.6%和0.8%）和胰岛素（不添加胰岛素、100 ng/mL和1000 ng/mL）浓度的无血清培养基中培养6天。

结果

在葡萄糖浓度高于0.2%时，胰岛素刺激VSMC生长（无添加胰岛素的0.4%葡萄糖培养基中细胞数为13,073±336个/mL，添加100 ng/mL胰岛素的0.4%葡萄糖培养基中细胞数为16,536±1175个/mL，添加1000 ng/mL胰岛素的0.4%葡萄糖培养基中细胞数为17,500±808个/mL，无添加胰岛素的0.6%葡萄糖培养基中细胞数为14,167±1062个/mL，添加100 ng/mL胰岛素的0.6%葡萄糖培养基中细胞数为18,984±1265个/mL，添加1000 ng/mL胰岛素的0.6%葡萄糖培养基中细胞数为20,450±1523个/mL，无添加胰岛素的0.8%葡萄糖培养基中细胞数为15,853±1650个/mL，添加1000 ng/mL胰岛素的0.8%葡萄糖培养基中细胞数为26,302±1919个/mL；与无添加胰岛素的葡萄糖组相比，P<.05）。无论培养基中的胰岛素浓度如何，葡萄糖在浓度达到0.2%时刺激VSMC增殖（与基线相比，0.1%和0.2%葡萄糖时生长分别高出42%和117%，P<.05）。胰岛素（1000 ng/mL）和葡萄糖（0.8%葡萄糖）浓度均最高的组生长最为显著（26,302±1919个/mL；P<.05）。