归芪益元膏对12C6+束辐射旁效应大鼠免疫功能的影响

Effects of Guiqi Yiyuan Ointment on Immune Function in 12C6+Beam Bystander-effect Rats

目的 观察归芪益元膏对12C6+束辐射旁效应大鼠免疫功能的影响及作用机制。方法 60只Wistar大鼠随机分为空白组、模型组、阳性对照组和中药低、中、高剂量组,每组10只。中药低、中、高剂量组予归芪益元膏1.2、2.4、4.8 g/kg灌胃,空白组、模型组、阳性对照组予等量生理盐水灌胃,共14 d;阳性对照组造模前30 min腹腔注射氨磷汀。第15日除空白组外,其余各组右肺予4 Gy12C6+束单次照射,照射后各组继续灌胃3 d。观察大鼠一般状况,检测外周血白细胞(WBC)、红细胞(RBC)、血红蛋白(Hb)、血小板(PLT)水平,ELISA检测血清白细胞介素(IL)-4、γ干扰素(IFN-γ)含量,计算肺、脾脏器指数,HE染色观察左肺、脾组织病理形态,流式细胞仪检测脾CD4+、CD8+T淋巴细胞比例,qPCR检测左肺、脾组织T-bet、GATA-3 mRNA表达。结果 与空白组比较,模型组大鼠外周血WBC降低,RBC、Hb、PLT升高,血清IL-4、IFN-γ含量降低;左肺、脾脏器指数明显降低,肺泡变形,肺泡壁变厚,肺泡腔缩小,肺组织明显出血,脾组织结构破坏严重,脾窦腔变窄,脾索出现纤维性增生;模型组脾CD4+T淋巴细胞比例和CD4+/CD8+比值降低、CD8+T淋巴细胞比例升高,左肺、脾组织T-bet mRNA表达降低,GATA-3 mRNA表达升高,差异均有统计学意义(P<0.05)。与模型组比较,阳性对照组和中药中、高剂量组大鼠外周血WBC升高,RBC、Hb、PLT降低,血清IL-4、IFN-γ含量升高;中药高剂量组左肺、脾脏器指数上升;各给药组肺、脾组织病理损伤均有不同程度减轻,阳性对照组和中药高剂量组脾CD4+T淋巴细胞比例和CD4+/CD8+比值升高,CD8+T淋巴细胞比例降低,左肺、脾组织T-bet mRNA表达升高,GATA-3 mRNA表达降低,差异均有统计学意义(P<0.05)。结论 归芪益元膏对12C6+束辐射旁效应大鼠肺、脾损伤有一定改善作用,其机制与调节CD4+、CD8+T淋巴细胞比例及T-bet、GATA-3表达,恢复细胞免疫功能有关。

Objective To observe the effects and mechanism of Guiqi Yiyuan Ointment on immune function in 12C6+beam bystander-effect rats.Methods Totally 60 Wistar rats were randomly divided into blank group,model group,positive control group,and TCM low-,medium-,and high-dosage groups,with 10 rats in each group.TCM low-,medium-,and high-dosage groups were given 1.2,2.4,and 4.8 g/kg Guiqi Yiyuan Ointment for gavage,and the blank group,model group and positive control group were given equal dose of sodium chloride solution for gavage for 14 days.Positive control group received intraperitoneal injection of amifostine 30 minutes before modeling.On the 15th day, except the blank group, the other groups were given 4 Gy 12C6+ beam of single irradiation on the rightlung. After irradiation, each group was given gavage for 3 days. The general condition of rats was observed,peripheral blood leukocyte (WBC), red blood cell (RBC), hemoglobin (Hb), platelet (PLT) levels were detected,ELISA was used to detect the contents of serum interleukin (IL)-4 and interferon-γ (IFN-γ), lung and spleen organindices were calculated, HE staining was used to observe the pathological morphology of left lung and spleen tissues,and flow cytometry was used to detect the proportion of CD4+ and CD8+ T lymphocytes in the spleen, qPCR wasused to detect the expressions of T-bet and GATA-3 mRNA in right lung and spleen tissues. Results Compared withthe blank group, the model group rats showed a decrease in peripheral blood WBC, an increase in RBC, Hb, PLT, andan decrease in serum IL-4 and IFN-γ contents;the organ indices of left lung and spleen was significantly reduced, thealveoli were deformed, the alveolar walls became thicker, the alveolar cavities shrank, the lung tissue wassignificantly bleeding, the spleen tissue structure was severely damaged, the splenic sinus cavity became narrower,and the splenic cord showed fibrous hyperplasia;the proportion of CD4+ T lymphocytes and CD4+/CD8+ ratio inspleen tissue of the model group decreased, while the proportion of CD8+ T lymphocytes increased, the expression ofT-bet mRNA in the left lung and spleen tissues decreased, while the expression of GATA-3 mRNA increased, withstatistical significance (P<0.05). Compared with the model group, the positive control group and TCM medium- andhigh-dosage groups showed an increase in peripheral blood WBC, an decrease in RBC, Hb, PLT, an increase inserum IL-4 and IFN-γ contents;the index of left lung and spleen organs increased in TCM high-dosage group;thepathological damage of left lung and spleen tissues in each treatment group alleviated to varying degrees, while theproportion of CD4+ T lymphocytes and CD4+/CD8+ ratio in spleen tissue of the positive control group and TCM highdosagegroup increased, the proportion of CD8+ T lymphocytes decreased, the expression of T-bet mRNA in left lungand spleen tissues increased, and the expression of GATA-3 mRNA decreased, with statistical significance (P<0.05).Conclusion Guiqi Yiyuan Ointment can alleviate the lung and spleen injury of rats induced by 12C6+ beam radiation.Its mechanism is related to regulating the proportion of CD4+, CD8+ T lymphocytes and the expression of T-bet andGATA-3, and recovering cellular immune function.