糖通方对体外培养糖尿病大鼠髂动脉血管平滑肌细胞增殖的影响

Effect of Tangtong Recipe on the Proliferation of Cultured Iliac Arterial Smooth Muscle Cells of Diabetic Rats in vitro

目的用血清药理学与分子生物学相结合的方法,探讨糖通方对糖尿病大鼠模型血管平滑肌细胞增殖的抑制作用。方法选择纯种新西兰家兔,每天灌服不同剂量(18、13、8mL/kg)的糖通方中药制剂,制备含有不同药物浓度的兔血清;用组织块种植法体外培养糖尿病大鼠模型的髂动脉平滑肌细胞(VSMC)并用不同浓度含药血清进行干预;用同位素液闪测定氚标记胸腺嘧啶脱氧核糖核苷(3H-TDR)掺入量,检测含药血清对平滑肌细胞增殖的影响;流式细胞仪测定细胞周期。结果各糖通方含药血清组呈浓度依赖性地表现为3H-TDR掺入量显著减少,3个含药血清组3H-TDR掺入量均明显低于正常血清对照组(P<0.01);流式细胞仪测定各含药血清组处于S和G2/M期细胞百分比显著低于正常血清对照组(P<0.01)。结论糖通方含药血清抑制体外培养糖尿病大鼠VSMC增殖,且抑制效应呈浓度依赖性。

Objective To explore the inhibition of Tangtong Recipe （TTR） on proliferation of vascular smooth muscle cells （VSMCs） in rat model of diabetes mellitus using seropharmacological and modern molecular biological technologies. Methods The drug-serum of TTR was prepared by feeding pure-breed New Zealand rabbits at various doses （18 mL/kg, 13 mL/kg, and 8 mL/kg） of TTR, and used to intervene the VSMCs get from the lilac artery of diabetic model rats cultured by tissue block plantation. The impact of drug-serum on VSMCs proliferation was estimated through measuring tritiated thymine deoxy-ribonucleotide （3H-TDR） incorporation using liquid-scintillation detector, and the cell cycle was detected using flow cytometry. Results The 3H- TDR incorporation in cells after intervention with various doses of TTR drug-serum were reduced significantly, in a concentration-dependent manner, to the level lower than that in the normal serum control group （ P 〈 0.01 ）. And the percentage of S and G2/M phase cells in the drug-serum treated groups also were markedly lower than that in the normal control group （ P 〈 0.01 ）. Conclusion Drug-serum of TTR could inhibit the proliferation of VSMC of diabetic rat in vitro in a concentration-dependent manner.