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背景:他汀类药物可以通过阻断甲羟戊酸的合成,抑制某些膜联结蛋白的翻译后修饰,从而阻断某些细胞内信号传导通路,抑制多种细胞的增殖。目的:探讨辛伐他汀对肺成纤维细胞的增殖、胶原合成及基质金属蛋白酶2分泌的影响。设计:完全随机设计,对照实验。单位:中国协和医科大学阜外心血管病医院器官移植研究室。材料:实验于2004-06/2004-10在中国协和医科大学北京协和医院心内科实验室完成。培养的新生SD大鼠的肺成纤维细胞,根据培养液中加入的干预药物浓度不同而分为辛伐他汀0,1,5,10,50μmol/L及辛伐他汀50μmol/L+甲羟戊酸200μmol/L组。方法:用消化法培养新生SD大鼠的肺成纤维细胞,给予不同浓度的辛伐他汀干预。四氮唑蓝比色法检测细胞增殖,细胞免疫组化法测定细胞胶原的合成,酶联免疫吸附法测定细胞培养上清液基质金属蛋白酶2的含量。主要观察指标:不同浓度辛伐他汀及辛伐他汀加甲羟戊酸干预后肺成纤维细胞增殖和胶原合成能力,以及基质金属蛋白酶2分泌量。结果:①辛伐他汀5,10,50μmol/L组四氮唑蓝比色A490值,肺成纤维细胞表达Ⅰ,Ⅲ型胶原的平均吸光度值(A值)及培养上清液中的基质金属蛋白酶2含量明显低于辛伐他汀0μmol/L组(0.520±0.010,0.334±0.011,0.260±0.012,0.111±0.011;0.508±0.011,0.324±0.014,0.232±0.015,0.083±0.015;0.445±0.017,0.305±0.015,0.216±0.015,0.068±0.012;0.561±0.013,0.361±0.012,0.289±0.012,0.140±0.013,t=3.359～8.111,P<0.05～0.01)。②辛伐他汀50μmol/L+甲羟戊酸200μmol/L组四氮唑蓝比色A490值,肺成纤维细胞表达Ⅰ,Ⅲ型胶原的平均吸光度值(A值)及培养上清液中的基质金属蛋白酶2含量明显高于辛伐他汀50μmol/L组(0.567±0.015,0.354±0.014,0.283±0.012,0.138±0.011,t=4.715～10.950,P<0.01)。结论:辛伐他汀能抑制肺成纤维细胞增殖和胶原合成,并可减少基质金属蛋白酶2的分泌,抑制肺成纤维细胞黏附迁移功能,并可通过影响甲羟戊酸通路而具有抗细胞增殖作用。
BACKGROUND: Statins can block certain intracellular signaling pathways and inhibit the proliferation of many kinds of cells by blocking the synthesis of mevalonate and inhibiting the post-translational modification of some membrane-associated proteins. Objective: To investigate the effects of simvastatin on the proliferation, collagen synthesis and matrix metalloproteinase 2 secretion of lung fibroblasts. Design: completely random design, control experiment. SETTING: Organ Transplantation Laboratory, Fuwai Hospital, Peking Union Medical College. MATERIALS: The experiment was performed at the Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College from June 2004 to October 2004. The cultured lung fibroblasts of neonatal SD rats were divided into simvastatin 0, 1, 5, 10, 50 μmol / L and simvastatin 50 μmol / L + mevalonate according to the concentration of the intervention drug in the culture solution 200μmol / L group. Methods: Pulmonary fibroblasts of neonatal SD rats were cultured by digestion, and different concentrations of simvastatin were given. Cell proliferation was measured by tetrazolium blue colorimetric assay, cell collagen synthesis was measured by immunohistochemistry and matrix metalloproteinase 2 concentration by enzyme-linked immunosorbent assay (ELISA). MAIN OUTCOME MEASURES: Proliferation and collagen synthesis of lung fibroblasts and secretion of matrix metalloproteinase 2 after intervention with simvastatin and simvastatin plus mevalonate. Results: (1) The average absorbance value (A value) and the average absorbance value (A value) of type Ⅰ and Ⅲ collagen in A4, 10 and 50μmol / L simvastatin groups, Protease 2 content was significantly lower than that of simvastatin 0μmol / L group (0.520 ± 0.010,0.334 ± 0.011,0.260 ± 0.012,0.111 ± 0.011,0.508 ± 0.011,0.324 ± 0.014,0.232 ± 0.015,0.083 ± 0.015; 0.445 ± 0.017, 0.305 ± 0.015,0.216 ± 0.015,0.068 ± 0.012; 0.561 ± 0.013,0.361 ± 0.012,0.289 ± 0.012,0.140 ± 0.013, t = 3.359 ~ 8.111, P <0.05 ~ 0.01). ② Simvastatin 50 micromol / L + mevalonate 200 micromol / L group, the A490 value of A480, the average absorbance value (type A) of type Ⅰ and type Ⅲ collagen in lung fibroblasts and matrix in culture supernatant The content of metalloproteinase 2 was significantly higher than that of simvastatin 50μmol / L group (0.567 ± 0.015,0.354 ± 0.014,0.283 ± 0.012,0.138 ± 0.011, t = 4.715 ~ 10.950, P <0.01). Conclusion: Simvastatin can inhibit lung fibroblast proliferation and collagen synthesis, and can reduce the secretion of matrix metalloproteinase 2, inhibit the adhesion and migration of lung fibroblasts, and may have an anti-cell proliferation effect by affecting the mevalonate pathway .