The Experimental Study Of Yiqi Huoxue Recipe On The Prevention Of Radiation-Induced Lung Injury In Rats

To study the effect of the Yiqi Huoxue Recipe on the expression of TGF-β1, TNF-α, ET-1,γ-IFN in rats with radiation-induced lung injury, and clarify the effective impact of Yiqi Huoxue Recipe on preventing the lung radioactive dandification, the expression of TGF-β1 protein and TNF-αprotein in human lung were examined by immunohistochemistry (ABC) method. Meanwhile, Microarray technique was also used to screen the susceptibility gene in radiation-induced lung injury, and finally to clarify the mechanism of it. The effect of Yiqi Huoxue Recipe on susceptibility gene expression of the radiation-induced lung injury can provide certain some scientific evidence for the clinical use of Yiqi Huoxue Recipe.70 cases of healthy rats (35 males and 35 females) were raised in separated cages. After feed in SPF laboratory for three days, those rats were divided into three groups at random by 3:3:1: namely radiational and the traditional Chinese medicines(TCM) group(A, n=30), simple exposure group(B, n=30) and control group(C, n=10). Rats in group A and group B were irradiated on the right lungs with 6MV-X Linear Accelerator. Before radiating, the rats were anesthetized with sodium pentobarbital 0.04g.Kg-1 by intraperitoneal injection and put on the platform; their limbs were fixed under Simulator with regulus lead blocking the left lung and mediastinum. The irradiation area is 3cm×3cm, one session every week for 5Gy and the total irradiation dose was 30Gy, awaking naturally after irradiation. Rats in Group C were only anesthetized without any irradiation. From the very day of irradiation, every rat in group A was treated with Yiqi Huoxue Recipe of 1mL per day, while the other two groups were given normal saline 1ml instead. Six rats, randomly selected from Group A and B each time at the 4^th 6^th 8^th 12^th 26^th weekend after first radiation, were executed immediately after intraperitoneal anesthesia. At the same time, six rats were also executed in group C at the end of 4 th week and two rats were needed for comparison at 6^th 12^th weekend . After executing the rats, serum were measured with enzyme-linked immunosorbent assay, right lung tissue were examined by using immunohistochemical methods, and the right lung tissue at the 6^th 12^th weekend were also taken for microarray examination. The changes of TGF-β₁,TNF-α,ET-1,γ-IFN in serum at different timepiece were measured with observing the dynamic growth of TGF-β1,TNF-αprotein expression.1. The serum level of TGF-β1,TNF-α,ET-1in those three groups began to increase after treatment at the 4^th week, reached the peak at the 12 th week and decreased slightly at the 26^th week; The expression levels of protein of the TGF-β₁,TNF-α,ET-1in group A were lower than those in group B(P<0.05). Yetγ-IFN began to decrease at the 4th week, decreased rapidly at the 12 th week and restored gradually by the 26^th week. The expression ofγ-IFN in group A in the serum were normally higher than those in group B(P<0.05).2. The protein expression of TGF-β₁,TNF-αin the lung tissues show that: the average of IOD of positive cell reactive material from the 4th week in both group A and group B began to increase and was higher than those in the control group, reached the peak at the 12^th week and slightly declined at the 26th week. But the average IOD value of group A were lower than that of group B(P<0.05) all time.3. Compared the rats in group B which were executed at the end of the 6th week with the rats in group C, one can see that among those differentially expressed genes, there were 669 up-regulated genes and 381 down-regulated genes. Compared the rats in group B which were executed at the end of the 12th week with those in the group C, the genes expressed differentially were screened including 833 up-regulated genes and 1217 down-regulated genes. The up-regulated genes related with radiation-induced lung injury were CTHRC, EDN1, IGF1, RAD, Bcl-2, MDM2, XRCC5, ERCC5. And the down-regulated genes included FGFR3, ERO1L, UNC5, STK4, ATRX, TP53, NFKB, FAS.4. Compared the executed rats in group A with those in group B at the 6^th weekend, the genes expressed differentially were choose including 936 up-regulated genes and 752 down-regulated genes. Rats which were executed at the 12^th weekend explained that those differentially expressed genes consist of 169 up-regulated genes and 183 down-regulated genes. Yiqi Huoxue Recipe has certain regulating effect on radiation-induced lung injury related to sensitivity genes CTHRC, FGFR3, EDN1, ERO1L.By inhibiting the expression of TGF-β₁, TNF-α, ET-1 and promoting the secretion ofγ-IFN, Yiqi Huoxue Recipe can relieve early radiated inflammation response and late radiated pulmonary fabrosis and has good effect on the prevention of radiation-induced lung injury. The genes of CTHRC1, FGFR3, EDN1, and ERO1L may be closely concerned with the cause of radiation-induced lung injury. Through regulating the expression of sensitivity genes CTHRC1,FGFR3,EDN1,ERO1L related to radiation-induced lung injury, Yiqi Huoxue Recipe paly a preventive role on radiation-induced lung injury.