谭潇;董宪喆;郭代红;王石;李牧函;赵润清;刘屏;
TAN Xiao;DONG Xian-zhe;GUO Dai-hong;WANG Shi;LI Mu-han;ZHAO Run-qing;LIU Ping;Department of Clinical Pharmacology,General Hospital of PLA;Department of Pharmacy,the 401 Hospital of PLA;Department of Pharmaceutical Care,General Hospital of PLA;

• 1:解放军总医院临床药理研究室
• 2:解放军401医院药剂科
• 3:解放军总医院药品保障中心

摘要(Abstract)：

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),iv)、鸡血藤组(生药10,20,40 g·kg^(-1))、儿茶素组(50,100,200 mg·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 effect and mechanism of ethanol extracts from Spatholobus suberectus and its active component catechin,ICR mice were exposed to 6Gy irradiation and randomly divided into normal group,model group,positive control group( amifostine,43. 6 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 effect and mechanism of ethanol extracts from Spatholobus suberectus and its active component catechin,ICR mice were exposed to 6Gy irradiation and randomly divided into normal group,model group,positive control group( amifostine,43. 6 mg·kg^(-1),iv 30 min before irradiation),SSD group( 10,20,40 g·kg(-1),iv 30 min before irradiation),SSD group( 10,20,40 g·kg^(-1)) and catechin group( 50,100,200 mg·kg(-1)) and catechin group( 50,100,200 mg·kg^(-1)). The mice were administered the appropriate drugs once a day after irradiation for 28 consecutive days. Blood samples were collected from the tail end and the number of peripheral blood cells was counted before irradiation and on day 1,3,7,14,21 and 28 using a microcell counter. Changes of thymus and spleen index of mice on day 7 were observed. The serum SOD,GSH-Px activity and MDA level were detected by the colorimetric method. The colony forming ability of bone marrow hematopoietic progenitor cells on day 7was detected by semi solid culture method. The HE staining was adopted to observe the pathological changes. The apoptosis of bone marrow cells was detected by flow cytometry. The expression of cleaved caspase-3 and Bax of bone marrow cells were measured separately by western-blotting and immunohistochemistry method. SSD and catechin can both significantly revert the irradiated-induced decline in hematological parameters( RBC,WBC,PLT,Hb),improve thymus and spleen index,significantly enhance serum SOD and GSH-Px activity and decrease the MDA level. The proliferation and differentiation of hematopoietic progenitor cells in bone marrow were promoted,the apoptosis of bone marrow cells was significantly up-regulated and the expression of cleaved caspase-3 and Bax was significantly reduced in SSD and catechin group. SSD and catechin have significant anti-radiation effect and its mechanism may be related to hematopoietic promoting,antioxidant and anti-apoptotic effects.

关键词(KeyWords)： 鸡血藤;儿茶素;辐射损伤;外周血象;造血祖细胞;细胞凋亡
Spatholobus suberectus;catechin;irradiation injury;peripheral blood;hemopoietic progenitor cell;cell apoptosis

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81373781)

作者(Authors): 谭潇;董宪喆;郭代红;王石;李牧函;赵润清;刘屏;
TAN Xiao;DONG Xian-zhe;GUO Dai-hong;WANG Shi;LI Mu-han;ZHAO Run-qing;LIU Ping;Department of Clinical Pharmacology,General Hospital of PLA;Department of Pharmacy,the 401 Hospital of PLA;Department of Pharmaceutical Care,General Hospital of PLA;

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