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研究鸡血藤乙醇提取物及其活性成分儿茶素的抗辐射作用,并探讨初步作用机制。采用6Gy60Coγ射线一次性全身照射ICR小鼠造成亚急性辐射损伤模型,随机分为正常组、模型组、阳性对照组(氨磷汀,43.6 mg·kg~(-1),iv)、鸡血藤组(生药10,20,40 g·kg~(-1))、儿茶素组(50,100,200 mg·kg~(-1)),连续ig给药28 d,每日1次。检测给药前1 d及给药1,3,7,14,21,28d后小鼠外周血WBC,RBC,PLT计数和HGB含量;观察给药7 d后小鼠胸腺及脾脏指数的变化;比色法测定给药7 d血清SOD,GSH-Px活性和MDA水平;采用半固体培养基测定给药7 d后骨髓造血祖细胞集落形成能力;HE染色观察给药7,14 d股骨骨髓病理学改变;流式细胞术检测给药7 d后骨髓细胞的凋亡,Western blot测定骨髓细胞中cleaved caspase-3和Bcl-2的表达,免疫组化法检测股骨骨髓中Bax的表达。结果表明,鸡血藤醇提物和儿茶素均可促进辐射小鼠外周血象的恢复,增加免疫器官胸腺和造血器官脾脏的脏器指数,提高血清抗氧化酶SOD和GSH-Px活性、降低MDA水平,促进骨髓造血祖细胞的增殖分化;增加骨髓腔内细胞数量、减轻立体支架结构损伤,改善造血微环境;抑制骨髓细胞凋亡,使凋亡相关蛋白Bcl-2的表达显著上调,cleaved caspase-3和Bax的表达显著下调。鸡血藤醇提物和儿茶素对亚急性辐射损伤小鼠有明显的保护作用,其机制与促造血、抗氧化、抗凋亡作用有关。
To study the anti-radiation effects of ethanol extract and the active ingredient catechin, and to explore the primary mechanism. The model of acute sub-acute radiation injury was induced by 6Gy60Coγ-ray irradiation in ICR mice. The models were divided into normal group, model group, positive control group (amifostine, 43.6 mg · kg -1, iv) (10, 20, 40 g · kg -1) and catechin group (50, 100, 200 mg · kg -1) were administrated continuously for 28 days once daily. The levels of WBC, RBC, PLT and HGB in the peripheral blood of mice were detected on the 1st day before administration and on the 1st, 3rd, 7th, 14th, 21st and 28th days after administration. Changes of thymus and spleen indexes in mice were observed 7 d after administration. Serum levels of SOD, GSH-Px and MDA were measured by colorimetric method. The hematopoietic progenitor cell colony formation ability of bone marrow was measured by semi-solid medium for 7 days. HE staining was used to observe the effect of 7,14 d femoral bone marrow disease Neutrophil apoptosis was detected by flow cytometry. The expression of cleaved caspase-3 and Bcl-2 in bone marrow cells was detected by Western blot. The expression of Bax in bone marrow of femur was detected by immunohistochemistry. The results showed that the ethanol extract and catechin could promote the recovery of peripheral blood, increase the organ index of thymus and hematopoietic organ of irradiated mice, increase the activities of serum antioxidant enzymes SOD and GSH-Px and decrease MDA level, promote the proliferation and differentiation of bone marrow hematopoietic progenitor cells; increase the number of cells in the marrow cavity, reduce the structural damage of stents and improve the hematopoietic microenvironment; inhibit the apoptosis of bone marrow cells, the expression of apoptosis-related protein Bcl-2 was significantly increased, cleaved The expression of caspase-3 and Bax was significantly down-regulated. Alcohol extracts and catechins have significant protective effect on sub-acute radiation-induced injury in mice. The mechanism is related to promoting hematopoietic, anti-oxidation and anti-apoptotic effects.