金属硫蛋白抑制同型半胱氨酸诱导的大鼠血管成纤维细胞激活  

Metallothionein inhibits rat vascular fibroblasts activation induced by homocysteine

作　　者：王冬艳[1] 郭杰[1] 李夏[2] 李扬[2] 唐朝枢[2] 李菊香[2] 

机构地区：[1]哈尔滨医科大学附属二院整形外科,黑龙江哈尔滨150086 [2]北京大学医学部生理学系,北京100083

出　　处：《中国病理生理杂志》2003年第6期773-777,共5页Chinese Journal of Pathophysiology

基　　金：黑龙江省攻关课题 (G99C2 0 - 10 - 2 )

摘　　要：目的 :观察外源性金属硫蛋白 (MT)和ZnCl2 诱导内源性MT生成对同型半胱氨酸 (HCY)激活大鼠血管成纤维细胞增殖的影响 ,以探讨MT拮抗HCY血管损伤的可能机理。方法 :采用 [3 H]-胸腺嘧啶 ([3 H]-TdR)和[3 H]-脯氨酸 ([3 H]-Pro)掺入法测定细胞增殖及胶原生成 ,自动生化分析仪测定乳酸脱氢酶 (LDH)漏出量 ,台盼蓝排斥法计算细胞存活率 ,并用 [10 9Cd]-血红蛋白饱和法检测细胞MT的含量。结果 :HCY 5 0、10 0、5 0 0 μmol/L呈浓度依赖性刺激血管成纤维细胞增殖 ,胶原合成和分泌 (P <0 0 5或P <0 0 1) ,5 0 0 μmol/LHCY使细胞LDH漏出量增多 ,细胞存活率降低。MT 1× 10 -5mol/L、1× 10 -4mol/L与 5 0 0 μmol/LHCY共孵育组与单纯HCY组比较 ,[3 H]-TdR、[3 H]-Pro掺入、胶原分泌降低 ,LDH漏出量减少 (P <0 0 5或P <0 0 1)。ZnCl2 预处理诱导细胞MT含量增加 (P <0 0 5 ) ,而ZnCl2 处理与HCY 5 0、10 0、5 0 0 μmol/L共孵育组 [3 H]-TdR和 [3 H]-Pro掺入 ,胶原分泌、LDH漏出量都明显低于单纯HCY组细胞 ,且细胞存活率高于单纯HCY组 (P <0 0 5或P <0 0 1)。结论 :HCY刺激血管成纤维细胞增殖与胶原合成 ,MT无论外源应用或内源性诱导生成均能有效拮抗HCY的上述作用。MT对HCY作用的拮抗效应可能是其血管细?AIM: To study the effects of exogenous metallothionein (MT) and ZnCl 2-induced MT production on biological action of homocysteine(HCY)in vascular fibroblasts METHODS: -TdR, -Pro incorporation and LDH leakage were measured, the cellular viabilities were calculated by trypan blue exclusion test and the intracellular contents of MT were assayed by [ 109 Cd]-hemoglobin saturation method in cultured rat vascular fibroblasts. RESULTS: Proliferation, collagen production of vascular fibroblasts in HCY-treated group were significantly increased compared with control group in a concentration-depedant manner. HCY (500 μmol/L) increased LDH leakage and decreased the cellular viabilities ( P< 0 05 or P< 0 01). -TdR incorporation,-Pro incorporation, collagen secretion and LDH leakage were all decreased in MT (1×10 -5 mol/L, 1×10 -4 mol/L) plus HCY(500 μmol/L) incubated group, compared with HCY alone group, respectively ( P< 0 05 or P< 0 01). MT content in ZnCl 2 pretreatment group was increased compared with control group. Proliferation,collagen production and LDH leakage in HCY group pretreated with ZnCl 2 were decreased whereas the cellular viabilities were increased compared with HCY alone group. CONCLUSIONS: The results shows that HCY induces proliferation and collagen production of vascular fibroblasts. Both exogenous MT and endogenous MT induced by ZnCl 2 inhibite the above-mentioned effects of HCY on vascular fibroblasts. MT inhibites vascular fibroblast activation induced by HCY, which may be related to its vascular protection.

关 键 词：金属硫蛋白 高胱氨酸 成纤维细胞 大鼠 

分 类 号：R363[医药卫生—病理学]