敦煌医方大补脾汤对放射性肠水肿大鼠的防护作用研究

A study on the protective effect of the Dunhuang medical formula Dabupi Decoction on rats with radiation-induced intestinal edema

关键词:  关键词：敦煌医方  大补脾汤  放射性肠水肿  防护作用

Key words:  Dunhuang medical formula  Dabupi Decoction  radiation- induced intestinal edema

摘要:

摘要：目的：探明敦煌医方大补脾汤对放射性肠水肿的防治作用及可能机制，为临床放射性肠水肿的防治及敦煌医方的推广提供实验依据。方法：实验分为3组：空白对照组、照射组和大补脾汤（6.8 g/kg）干预组。观察大鼠一般状况；苏木精-伊红染色观察结肠组织形态变化；透射电镜检测结肠组织超微结构变化；ROS染色检测结肠组织中ROS表达水平；激光共聚焦检测结肠组织中HIF-1α和AQP4表达水平；蛋白免疫印迹法检测结肠组织HIF-1α、AQP4、HOIL-1L、PKCζ、Occludin和Na+/K+-ATPase蛋白的表达水平。结果：HE染色结果表明：辐射组大鼠出现黏膜坏死脱落、黏膜及黏膜下水肿，大补脾汤干预组黏膜脱落和水肿现象减轻。透射电镜结果显示：辐射组细胞质基质肿胀，线粒体肿胀变形，扩张形成空泡化，大补脾汤干预组线粒体肿胀明显好转，胞质内大面积空泡现象明显改善。激光共聚焦结果表明：与空白组相比，辐射组ROS、HIF-1α和AQP4表达显著升高（P<0.05，P<0.01）；与辐射组相比，大补脾汤组ROS、HIF-1α和AQP4表达显著降低（P<0.05，P<0.01）。蛋白免疫印迹结果表明：与空白组相比，辐射组HIF-1α、AQP4和HOIL-1L蛋白表达显著升高（P<0.01），PKCζ、Occludin和Na+/K+-ATPase蛋白表达显著降低（P<0.01）；与辐射组相比，大补脾汤组HIF-1α、AQP4和HOIL-1L蛋白表达显著降低（P<0.01），PKCζ、Occludin和Na+/K+-ATPase蛋白表达显著升高（P<0.01）。结论：敦煌医方大补脾汤可以有效防护放射性肠水肿，其机制可能是通过抑制ROS、HIF-1α和AQP4的表达，促进PKCζ、Na+/K+-ATPase以及Occludin的表达来发挥作用。

Abstract:

ABSTRACT: OBJECTIVE To investigate the effect and possible mechanism of the Dunhuang medical formula Dabupi Decoction on the prevention and treatment of radiation-induced intestinal edema, and to provide an experimental basis for the prevention and treatment of clinical radiation-induced intestinal edema and the promotion of the Dunhuang medical formula. METHODS The experiment was divided into 3 groups: blank control group, irradiated group and the intervention group of Dabupi Decoction (6.8 g/kg). The general conditions of the rats were observed; morphological changes of colonic tissues were observed by hematoxylin-eosin staining; ultrastructural changes of colonic tissues were detected by transmission electron microscopy; ROS expression levels in colonic tissues were detected by ROS staining; HIF-1α and AQP4 expression levels in colonic tissues were detected by laser confocal; the expression levels of HIF-1α, AQP4, HOIL-1L, PKCζ, Occludin and Na+/K+-ATPase protein expression levels. RESULTS The HE staining results showed that the radiation group showed necrotic detachment of mucous membrane, mucous membrane and submucous membrane edema, while the mucous membrane detachment and edema were reduced in the Dabupi Decoction intervention group. The results of transmission electron microscopy showed that the cytoplasmic matrix of the radiation group was swollen and the mitochondria were swollen and deformed and expanded to form vacuolation, while the mitochondrial swelling in the Dabupi Decoction intervention group was significantly improved and the large intracytoplasmic vacuolation was significantly improved. Laser confocal results showed that the expression of ROS, HIF-1α and AQP4 was significantly higher in the radiation group compared with the blank group (P<0.05, P<0.01), while the expression of ROS, HIF-1α and AQP4 was significantly lower in the Dabupi Decoction intervention group compared with the radiation group (P<0.05, P<0.01). The results of protein immunoblotting showed that compared with the blank group, the expression of HIF-1α, AQP4 and HOIL-1L protein was significantly higher (P<0.01) and the expression of PKCζ, Occludin and Na+/K+-ATPase protein was significantly lower (P<0.01) in the radiation group; compared with the radiation group, the expression of HIF-1α, AQP4 and HOIL-1L protein expression was significantly lower (P<0.01) and PKCζ, Occludin and Na+/K+-ATPase protein expression was significantly higher (P<0.01) compared with the radiation group. CONCLUSION The dunhuang medical formula Dabupi Decoction can effectively protect against radiation-induced intestinal edema, and the mechanism may work by inhibiting the expression of ROS, HIF-1α and AQP4 and promoting the expression of PKCζ, Na+/K+-ATPase and Occludin.