归芪益元膏对重离子辐射诱发体内旁效应中Cyt-C，Bax及Bcl-2表达的影响

目的:探讨归芪益元膏对重离子辐射旁效应大鼠肺肾组织中细胞色素-C(Cyt-C)及Bax,Bcl-2基因表达的影响。方法:Wistar雄性大鼠随机分为7组,归芪益元膏组预先灌胃归芪益元膏2周(灌胃剂量为3.28 g·kg~(-1)·d~(-1)),正常组和辐射组灌胃等量生理盐水。2周后辐射组和归芪益元膏组大鼠右肺予2Gy~(12)C~(6+)离子束单次照射,正常组不予照射。照射后6,12,24 h处死各组大鼠,蛋白免疫印迹(Western blot),免疫组化及实时荧光定量聚合酶链式反应(Real-time PCR)检测各组大鼠右肺、左肺、左肾组织中Cyt-C,Bax,Bcl-2蛋白及mRNA表达。结果:右肺、左肺、左肾中Cyt-C,Bax mRNA表达在照射后6,12,24h均明显上调,而蛋白表达只在照射后24 h明显上调,与正常组比较,差异有统计学意义(P0.01);右肺中,Bcl-2蛋白及mRNA表达只在照射后24 h明显下调,而左肺、左肾中照射后6,12,24 h均明显下调,与正常组比较,差异有统计学意义(P0.01)。右肺、左肺、左肾中,归芪益元膏组Cyt-C,Bax mRNA表达在照射后6,12,24 h较辐射各组明显下调,归芪益元膏组Cyt-C,Bax蛋白表达在照射后24 h较辐射组明显下调,差异有统计学意义(P0.01);右肺中,归芪益元膏组Bcl-2 mRNA及蛋白表达在照射后24 h较辐射组明显上调,左肺、左肾中,归芪益元膏组Bcl-2 mRNA及蛋白表达在照射后6,12,24 h较辐射各组明显上调,差异有统计学意义(P0.01)。结论:重离子辐射大鼠右肺后,左、右肺和左肾组织中Cyt-C,Bax,Bcl-2基因表达异常,归芪益元膏通过延缓Bcl-2表达的下调而抑制Bax,Cyt-C表达的上调,从而起到预防肺肾细胞凋亡,发挥其对重离子辐射诱发体内旁效应损伤的防护作用。

Influence of Guiqi Yiyuangao on Expression of Cytochrome-C and Bax,Bcl-2 During Damage Caused by Heavy Ion Radiation Induced Bystander Effect in Vivo

Objective: To discuss the influence of Guiqi Yiyuangao on the genetic expression of Bax, Bcl-2 and cytochrome-C(Cyt-C) in lung and kidney of rats damaged by heavy ion radiation induced bystander effect. Method: The Wistar male rats were randomly divided seven groups, the group of Chinese medicine in advance was given Guiqi Yiyuangao by gavage for two weeks. Later the right lung of the rats of the radiation group and the group of Chinese medicine was radiated by ¹²C⁶⁺ ion once. At 6, 12, 24 h after radiation all groups of rats were executed. The amounts of protein and mRNA expression of Bax, Bcl-2 and Cyt-C in right lung, left lung and left kidney were examined with Western blot, immunohistochemistry and Real-time PCR. Result: Compared with the normal control group, the amount of mRNA expression of Bax and Cyt-C in the right lung, left lung and left kidney of the radiation group obviously increases at 6,12,24 h after radiation. While the amount of protein expression of Bax and Cyt-C of the radiation group obviously increases at 24 h after radiation. There is statistically significant difference(P<0.01). The amount of protein and mRNA expression of Bcl-2 in the right lung of the radiation group obviously weakens at 24 h after radiation. But in the left lung and left kidney of the radiation group it obviously weakens at 6, 12, and 24 h after radiation. There is statistically significant difference(P<0.01). Compared with the radiation group, the mRNA expression of Bax and Cyt-C of the Chinese medicine groups obviously down-regulates at 6,12,24 h after radiation. While the protein expression of the Chinese medicine group obviously down-regulates at 24 h after radiation. There is statistically significant difference(P<0.01). Compared with the radiation group, the expression of mRNA and protein of Bcl-2 in the right lung of the Chinese medicine group obviously up-regulates at 24 h after radiation. Compared with the radiation group, the expression of mRNA and protein of Bcl-2 in the left lung and left kidney of the Chinese medicine group obviously up-regulates at 6,12,24 h after radiation. There is statistically significant difference(P<0.01). Conclusion: It indicates that there are abnormal expressions of Bax, Bcl-2 and Cyt-C of lung and kidney in rats damaged by heavy ion radiation induced bystander effect. By delaying the down-regulation of the expression of Bcl-2, controlling the up-regulation of the expression of Bax and Cyt-C, Guiqi Yiyuangao can prevent the lung and kidney cell apoptosis and defend the damage caused by heavy ion radiation induced bystander effect in vivo.