淫羊藿苷体内抑瘤作用及其机制  

The in vivo anti-tumor effects of Icariin and its mechanisms

作　　者：李翠玲[1] 张玲[1] 顾洪涛[1] 王军[1] 毛海婷[1] 杨尚军[2] 温培娥[1] 

机构地区：[1]山东省医学科学院基础医学研究所 [2]山东省医学科学院药物研究所,济南250062

出　　处：《中国肿瘤生物治疗杂志》2007年第2期137-142,共6页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金主任基金资助项目(No.30540041)

摘　　要：目的:探讨淫羊藿苷(icariin,ICA)对肝癌细胞H22荷瘤小鼠体内抑瘤作用及其机制。方法:肝癌细胞株H22右腋皮下接种C57BL/6小鼠,建立小鼠体内荷移植瘤模型。接种H22细胞后24h,用高、中、低不同剂量(9、4.5、2.25mg/ml)的ICA和环磷酰胺(cyclophosphamide,Cy)(30mg/kg)进行局部注射治疗,待荷瘤对照组肿瘤长至1g左右,各组小鼠称重,处死。留自凝血,取血清检测TNF-α;依次取脾脏及肿瘤组织称重;取1/2脾脏和肿瘤组织制成单细胞悬液,剩余的用10%中性甲醛溶液固定,做病理切片和H-E染色以及TUNEL实验;脾单细胞悬液用流式细胞术(FACS)检测CD3、CD4、CD8、DX5等抗原分子的表达;肿瘤单细胞悬液用AnnexinV/PI双染检测肿瘤细胞的早期与晚期凋亡情况,同时取适量的瘤细胞用70%乙醇固定以FACS检测肿瘤细胞周期各时相的分布情况。结果:Cy治疗组抑瘤率为67.16%,ICA高、中、低剂量治疗组抑瘤率分别为24.63%、32.84%、5.22%。AnnexinV/PI双染结果表明,Cy治疗组与ICA治疗组早期凋亡细胞率均高于荷瘤对照组[Cy组为(9.11±1.584)%,ICA组为(7.14±1.376)%,荷瘤对照组为(3.6±2.38)%]。TUNEL结果显示,ICA治疗组凋亡指数为(5.35±0.73)%,显著高于荷瘤对照组中的凋亡指数[(1.03±1.17)%](P<0.01)。正常小鼠血清中未检测到TNF-α的存在,而ICA治疗组及Cy治疗组中检测到微量TNF-α。病理检测表明,ICA中、高剂量组肿瘤组织中出现较多的灶状及小片状坏死,包膜处及肿瘤边缘可见瘤细胞坏死,包膜及肿瘤组织中可见少量的炎细胞(淋巴细胞与单核细胞)浸润。结论:ICA有显著的抑瘤作用,可能通过促进实体瘤细胞早期凋亡引起肿瘤组织坏死而起作用。Objective： To investigate the in vivo anti-tumor effects of Icarrin （ICA） on H22-bearing mice and the re-ated anti-tumor mechanisms. Methods： C57BL/6 mice were subcutaneously inoculated with H22 hepatocarcinoma cell line to establish H22-bearing mice model. 24 h after the inoculation, H22-bearing mice were treated with different doses of ICA （9.0 mg/ml, 4.5 mg/ml, and 2.25 mg/ml） and cyclophosphamide （30 mg/kg）. When the tumors in H22-bearing mice grew to 1 g, the mice were weighed, sacrificed and their spleens and tumors were harvested. The mice sera were also collected for determination of TNF-α. Half of the spleens and tumors were made into single-cell suspensions and the other half was fixed by 10% formaldehyde solution for pathology analysis and TUNEL detection. Flow cytometry technique was used to detect the expression of surface antigens CD3, CIM, CD8, and DX-5 in the spleen. Annexin- V/PI double staining method was used to analyze the early and late apoptosis of tumor single-cell suspension. 1.0 ×10^6 cells were used to analyze the cell cycle of the solid tumor cells by flow cytometry. Results： The tumor inhibition rate of cyclophosphamide was 67.16% （P 〈 0. 001 ）, and those of high-, medium- and low-dose ICA groups were 24.63% （P 〈 0.05 ）, 32.84% （P 〈 0.01 ） and 5.22%, respectively. Annexin V/PI double staining results indicated that the early apoptosis rate of tumor cells in cyclophosphamide group and medium-dose ICA group were higher than those in H22-bearing group [Cy （ 9. 11 ± 1. 584） %, ICA （7.14 ± 1. 376） %, H22-bearing group （3.6 ± 2.38） % ]. TUNEL result showed that apoptosis index in ICA-medium group was significantly higher than that in H22-bearing group [ （ 5.35 ± 0.73 ） % vs （ 1.03±1. 17）% , P 〈0.01 ]. No TNF-α was detected in normal control mice and only very low level of TNF-α was found in ICA treatment group and Cy group. Pathology study showed that there were severe necrosis in tumors of medium- and high-dose ICA groups, w

关 键 词：淫羊藿苷 抑瘤作用 细胞凋亡 细胞周期 

分 类 号：R730.5[医药卫生—肿瘤]