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目的:检测益气活血方干预前后大鼠放射性肺损伤组织基质金属蛋白酶2(matrix metalloproteinase-2,MMP2)和基质金属蛋白酶抑制剂2(tissue inhibitor of matrix metalloproteinase,TIMP2)表达的影响,了解该方剂的作用机制。方法:健康成年雌性SD大鼠120只,随机分为模型组30只、照射加益气活血方高剂量组30只、照射加益气活血方低剂量组30只、空白组30只。益气活血方高、低剂量组大鼠受照当天开始,用药量20,10 g·kg-1,每次ig益气活血方剂5 m L·kg-1,2次/d;模型组、空白组每次ig蒸馏水5 m L·kg-1,2次/d。4组大鼠分别于照射后第4,8,12,16,20周末随机抽取6只,处死并取其右肺组织保存。分别用反转录PCR技术(RT-PCR),蛋白质印迹法(Western-blotting)检测MMP-2/TIMP-2 mRNA及蛋白表达的动态变化。结果:模型组MMP-2 mRNA及蛋白表达于第12周达高峰,而TIMP-2 mRNA及蛋白表达在照射后第4周至20周期间出现逐渐升高的趋势,各时间点模型组与空白组相比均具有明显差异(P<0.01)。益气活血方高剂量组与模型组相比,各mRNA及蛋白表达趋势与模型组相似,MMP-2 mRNA及蛋白表达在12周前低于模型组,差异有统计学意义(P<0.05,P<0.01)。益气活血方低剂量组各项指标均有变化,趋势与模型组类似,差异没有统计学意义。结论:益气活血方高剂量在放射性肺损伤的各个时间段,从转录及翻译层面对MMP-2/TIMP-2的表达进行调整,使MMPs/TIMPs趋于平衡,从而减轻放射性肺损伤,这可能是该药防治放射性肺损伤的分子生物学机制。
Objective: To investigate the effect of Yiqi Huoxue Recipe on the expression of matrix metalloproteinase-2 (MMP2) and tissue inhibitor of matrix metalloproteinase (TIMP2) in radiation-induced lung injury in rats before and after intervention The mechanism of action. Methods: A total of 120 healthy female Sprague-Dawley rats were randomly divided into model group (n = 30), high dose irradiation group (n = 30) and low dose group (n = 30). Yiqihuoxue Fang high and low dose group rats started the day of irradiation, the dosage of 20,10 g · kg-1, each ig Yiqihuoxue Fang 5 m L · kg-1, 2 times / d model group, Blank group ig distilled water 5 m L · kg-1, 2 times / d. Four rats were randomly selected at the end of the 4th, 8th, 12th, 16th and 20th week after irradiation respectively. Six rats were sacrificed and their right lung tissues were preserved. The dynamic changes of MMP-2 / TIMP-2 mRNA and protein expression were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blotting respectively. Results: The expression of MMP-2 mRNA and protein peaked at the 12th week in the model group, while the expression of TIMP-2 mRNA and protein increased gradually from the 4th week to the 20th week after irradiation. At each time point, Compared with significant differences (P <0.01). Compared with the model group, the expression of mRNA and protein of Yiqi Huoxue Fang high-dose group was similar to that of the model group, and the expression of MMP-2 mRNA and protein was lower than the model group 12 weeks ago (P <0.05, P <0.01). Yiqihuoxue low-dose group indicators have changed, the trend is similar to the model group, the difference was not statistically significant. CONCLUSION: The high dose of Yiqi Huoxue regulates the expression of MMP-2 / TIMP-2 from the transcriptional and translational levels during various periods of radiation-induced lung injury, which leads to the balance of MMPs / TIMPs and thus the radiation-induced lung injury. It may be the drug’s molecular mechanism of prevention and treatment of radiation-induced lung injury.