黄芪注射液对放疗作用下食管癌大鼠抗肿瘤功能的影响及其机制研究

Effects of astragalus injection on antitumor function of esophageal cancer rats under radiotherapy and its mechanism

目的探讨黄芪注射液(AI)对放疗作用下食管癌大鼠抗肿瘤功能的影响及其机制。方法采用甲基苄基亚硝胺构建食管癌放射治疗模型,将64只Wistar大鼠按照随机数字表法分为4组:对照组、阳性对照组(EM+RT组)、放射治疗组(RT组)和治疗组(AI+RT组),每组各16只。其中对照组不接受任何治疗,RT组仅进行放射治疗,EM+RT组放射后给予不锈钢喂食针食管内灌注111.7 g/mL的食道混合物治疗,AI+RT组放射治疗后给予不锈钢喂食针食管内灌注大鼠100 g/mL AI。在放射后7、14、21和28 d,测定各组大鼠体重,并取食管组织进行组织病理学和免疫组织化学染色,进一步采用酶联免疫吸附试验(ELISA)法测定血清白细胞介素(IL)-6、IL-10和肿瘤坏死因子-α(TNF-α)。结果放射后14、21、28 d,与对照组相比,RT组、EM+RT组和AI+RT组大鼠体重显著降低,差异均有统计学意义(P<0.05);放射后14、21、28 d,与RT组比较,EM+RT组、AI+RT组体重显著降低,差异均有统计学意义(P<0.05)。放射后28 d,RT组大鼠黏膜上皮严重坏死脱落,基底细胞层出现多个空泡,部分病例有溃疡灶。AI治疗改善了黏膜上皮层的完整性,增加了固有层的厚度,并减少了中性粒细胞浸润。放射后7、14、21、28 d,与正常组相比,RT组碱性成纤维细胞生长因子(bFGF)及转化生长因子β1(TGF-β1)的阳性细胞数量显著增加,差异均有统计学意义(P<0.05);放射后14、21、28 d,与RT组比较,AI+RT组bFGF和TGF-β1的阳性细胞数量均显著降低,差异均有统计学意义(P<0.05)。放射后7、14、21、28 d,与正常组相比,RT组血清TNF-α、IL-6和IL-10水平显著升高,差异均有统计学意义(P<0.05);与RT组比较,AI+RT组、EM+RT组血清TNF-α、IL-6和IL-10水平显著降低,差异均有统计学意义(P<0.05)。结论黄芪注射液在食管癌大鼠放射治疗过程中发挥显著的治疗作用,其作用机制可能与改善机体细胞bFGF和TGF-β1表达及调节血清炎症因子水平有关,因此黄芪注射液可作用治疗食管癌放射性治疗后并发症的潜在替代方案。

Objective To investigate the effect of astragalus injection(AI)on the antitumor function of rats with esophageal cancer under radiotherapy and its mechanism.Methods A model of esophageal cancer radiotherapy was established by methylbenzylnitrosamine.And 64 Wistar rats were divided into 4 groups according to the random number table method:control group,radiotherapy group(RT),positive control group(EM+RT)and treatment group(AI+RT),16 rats in each group,and all groups were model rats.The control group did not receive any treatment,the RT group only received radiotherapy,the EM+RT group received 111.7 g/mL esophagus mixture with stainless steel feeding needle after radiotherapy,the AI+RT group received radiotherapy combined with stainless steel feeding needle intraesophageal perfusion of 100 g/mL AI in rats.At 14,21,and 28 days after radiation2,the body weight of the rats in each group was measured,and the esophageal tissue was taken for histopathological and immunohistochemical staining.The serum interleukin-6(IL-6),interleukin-10(IL-10)and tumor necrosis factor-α(TNF-α)were further determined by enzyme-linked immunosorbent assay(ELISA).Results At 14,21,and 28 days after radiation,compared with the control group,the body weight of the rats in RT group,EM+RT group and AI+RT group were significantly decreased,and the differences were statistically significant(P<0.05).At 14,21,and 28 days after radiation,compared with the RT group,the body weight of EM+RT group and AI+RT group were significantly decreased,and the differences were statistically significant(P<0.05).At 28 days after radiation,in the RT group,the mucosal epithelium was severely necrotic and shed,and multiple vacuoles appeared in the basal cell layer,and some cases had ulcers.AI treatment improves the integrity of the mucosal epithelial layer,increases the thickness of the lamina propria,and reduces neutrophil infiltration.At 7,14,21,and 28 days after radiation,compared with the normal group,the numbers of basic fibroblast growth factor(bFGF)and transforming growth factorβ1(TGF-β1)positive cells in the RT group were significantly increased,and the differences were statistically significant(P<0.05).At 7,14,21,and 28 days after radiation,compared with the RT group,the numbers of bFGF and TGF-β1 positive cells in the AI+RT group were significantly decreased,and the differences were statistically significant(P<0.05).At 7,14,21,and 28 days after radiation,compared with the normal group,the serum levels of TNF-α,IL-6 and IL-10 in the RT group were significantly increased,and the differences were statistically significant(P<0.05).Compared with RT group,the levels of serum TNF-α,IL-6 and IL-10 in AI+RT group and EM+RT group were significantly decreased,and the differences were statistically significant(P<0.05).Conclusion Astragalus injection plays a significant role in the radiation therapy of esophageal cancer rats,and its mechanism may be related to improving the expression of bFGF and TGF-β1 in body cells and regulating the level of serum inflammatory response.Therefore,astragalus injection can be a potential alternative for the treatment of complications after radiotherapy for esophageal cancer.