The Effect Of Yiqi Huoxue Recipe To Sensitive Genes Expression In Radioactive Lung Injury

To analyze the differential expression in the process of rats with radioactive pulmonary injury by digital gene expression profiling in order to find sensitive genes, Yiqi Huoxue Recipe is used to gavage the irradiated rats, then observe the effect of the mRNA and protein expression of the radiation-induced lung injury tissue, in order to analyze the mechanism of the drug from transcription and translation levels, and to procide an ontological bacsis of the durg promotion.90cases of healthy adult female rats are divided into three groups at random: namely simple exposure group(model group, n=30), radiational and traditional Chinese medicines group(Chinese medicine group, n=30), and blank control group(control group, n=30). Rats in model group and Chinese medicine group are irradiated on the right lung with6MV-X Linear Accelerator. Before radiating, the rats are anesthetized with7%chloral hydrate5·kg-1by intraperitoneal injection and put on the platform, with regulus lead blocking the left lung and mediastinum. The irradiation area is3cm×3cm,5Gy for every weeks one session and the total irradiation dose is30Gy, awaking naturally after irradiation. Rats in control group are only anesthetized without any irradiation. From the very day of irradiation, every rat in Chinese medicine group is treated with Yiqi Huoxue Recipe1ml every time, twice a day; while the other two groups are given distilled water1ml every time, twice a day. Six rats are randomly selected from the three groups each time at the4th,6th,8th,12th,26th weekend after first radiation. The right lung tissue of those rats which are executed immediately after intraperitoneal anesthesia and save in right way. Take part irradiated lung tissue of the model group, and normal lung tissue of the control group in4th,12th. Extract their total RNA, and give it to HuaDa gene company to do digital gene expression analysis. The rest lung tissue is dealt with RT-PCR to detect the dynamic change of mRNA expression of MMP-2, MMP-9, TIMP-1and TIMP-2in each time point. Detect the dynamic change of Protein expression of MMP-2, MMP-9, TIMP-1and TIMP-2with Western-blotting in each time point. Use SPSS17.0statistical software to do statistical analysis. The results of gene expression suggest that MMP-2, MMP-9, TIMP-1, TIMP-2genes may be sensitive genes for radioactive lung injury. The results of RT-PCR, Western-blotting and immunohistochemical suggest that the expression of MMP-2, MMP-9mRNA and protein appears an increase first and then decline trend from4th weekend to20th weekend after irradiation. The express peak of MMP-2mRNA and protein is in12th, and the express peak of MMP-2mRNA and protein is in16th. The expression of TIMP-1, TIMP-2mRNA and protein appears a rising trend from4th weekend to20th weekend after irradiation. The mRNA and protein expression trend is similar in Chinese medicine group and model group, but they have different change in different time point.We find that MMP-2, MMP-9, TIMP-1, TIMP-2genes may be sensitive genes for radioactive lung injury. The Yiqi Huoxue Recipe can adjust the expression of MMP-2, MMP-9, TIMP-1, TIMP-2from transcription and translation level in each period of radioactive lung injury. We can make MMPs/TIMPs getting balanced, so as to reduce the radioactive lung injury. This may be the molecular biology mechanism for this medicine to protect and control the radioactive lung injury.