当归红芪超滤膜提取物对12C6＋重离子束辐射致H22荷瘤小鼠免疫功能损伤的影响

Effects of ultra-filtration extract from Danggui and Hongqi on immune function injury induced by ^(12)C^(6+) heavy ion beam radiationin H_(22) tumorbearing mice

目的观察当归红芪超滤膜提取物抗^(12)C^(6+)重离子束辐射致H22荷瘤小鼠免疫功能损伤的作用。方法 H22细胞皮下注射法构建H22荷瘤小鼠模型,以检测血液白细胞数目为评价标准确定^(12)C^(6+)重离子束辐射致H22荷瘤小鼠免疫损伤最佳辐射剂量及当归红芪超滤膜提取物药物干预最佳剂量。实验分正常对照组、肿瘤模型组、单纯药物组(给予当归红芪超滤膜提取物6.72 g/kg)、单纯放射组(照射剂量为4Gy)、药物(给予当归红芪超滤膜提取物6.72 g/kg)+放射(照射剂量为4Gy)组。测定各组脾脏指数、胸腺指数;流式细胞仪技术分析各组小鼠血液T淋巴细胞亚群变化;ELISA法检测各组小鼠血清补体C3、干扰素-γ(IFN-γ)、白细胞介素-4(IL-4)含量;光镜下观察脾脏病理形态学改变。结果与肿瘤模型组比较,单纯放射组血液白细胞数目、脾脏指数、胸腺指数、血液CD3+、CD4+、CD8+T淋巴细胞数量、血清补体C3、IFN-γ、IL-4含量明显下降(P<0.05),CD4+/CD8+比值、IFN-γ/IL-4比值明显下降(P<0.05)。病理形态学显示:肿瘤模型组小鼠脾脏形态结构如常,多数脾窦扩张淤血,脾小体清晰,可见动脉周围淋巴鞘,巨核细胞多见,与正常对照组小鼠脾脏形态无明显差异;单纯放射组脾脏脾小体及红髓脾索萎缩,淋巴细胞明显减少,巨核细胞数量减少,纤维组织增生。与单纯放射组比较,药物+放射组血液白细胞数目、脾脏指数、胸腺指数、血液CD3+、CD4+、CD8+T淋巴细胞数量、血清补体C3、IFN-γ含量明显上升(P<0.05),CD4+/CD8+比值、IFN-γ/IL-4比值有明显上升(P<0.05),病理形态学损伤改变明显减轻。结论当归红芪超滤膜提取物有减轻^(12)C^(6+)重离子束辐射引起H22的荷瘤小鼠免疫功能损伤的作用。

Objective To observe the effects of ultra-filtration extract from Danggui （Radix Angelicae Sinensis, Chinese Angelica） and Hongqi （Radix Hedysair, Sweetvetch Root） on immune dysfunction caused by 12C6＋ heavy ion beam radiation in H22 tumor-bearing mice. Methods Tumor-bearing mice model was built by subcutaneously injecting H22 cells. Blood leukocytes count was used to determine the optimal radiation dose of immune injury induced by heavy ion beam irradiation and the optimal dose of ultra-filtration extract from Danggui and ttongqi. The mice were divided into normal control group, tumormodel group, drug intervention group （6.72 g/kg）, radiation group （4Gy）, and drug （6.72 g/kg） plus radiation （4Gy） group. Spleen index and thymus index were measured in each group; Changes of T lymphocyte in each group was detected by using flow cytometry; Levels of serum C3, IFN-γ and IL-4 were tested with ELLSA; Spleen pathological and morphological changes were observed under optical microscope. Results Compared with the model group, blood leukocyte count, spleen index, thymus index, levels of CD3 ＋ , CD4 ＋ and CD8 ＋ T lymphocytes, serum C3, IFN-γ and IL-4 in radiation group decreased significantly （ P 〈 0.05 ） , and ratio of CD4 ＋/CD8 ＋ and IFN-γ/IL-4 in radiation group de- creased significantly （P 〈 0.05 ）. Pathological morphology indicated that there was no significant differ- ence between the model group and normal control group. Spleen structure was normal; most of the splenic sinus dilated and developed blood stasis. Splenic corpuscle was clear. The lymphatie sheath around the artery was present, and macronuelear cells could be observed in the model group. Whereas in the radia- tion group, the spleen body and red pulp spleen cord shrank ; the number of lymphocyte and megakaryo- cytes reduced, and fibrous tissue proliferated. Compared with the radiation group, blood leukocytes count, spleen index, thymus index, the levels of CD3 ＋ , CD4 ＋ and CD8 ＋ T lymphocytes, serum C3, IFN-γ and IL-4 in drug plus radiation group increased significantly （P 〈 0.05 ） , and the ratio of CD4 ＋ / CD8 ＋ and IFN-γ/IL-4 in drug plus radiation group increased markedly （P 〈 0.05 ）. Compared with the radiation group, pathological changes of spleen in the drug plus radiation group alleviated significantly. Conclusions The ultra-filtration extract from Danggui and Hongqi can could alleviate the immune func- tion injury caused by 12C6＋ heavy ion beam radiation in H22 tumor-bearing mice.