川芎嗪对H22肝癌小鼠血管生成及KLF4/VEGF/ADAMTS1通路的影响  

Effect of Ligustrazine on Angiogenesis and KLF4/VEGF/ADAMTS1 Pathway in H22 Liver Cancer Mice

作　　者：张占栋 马瑞 李瑞芳[2] 李三强[2] 都婉红 ZHANG Zhandong;MA Rui;LI Ruifang;LI Sanqiang;DU Wanhong(Henan Vocational College of Tuina,Luoyang 471023,China;College of Basic Medicine and Forensic Medicine,Henan University of Science and Technology,Luoyang 471023,China)

机构地区：[1]河南推拿职业学院,河南洛阳471023 [2]河南科技大学基础医学与法医学院,河南洛阳471023

出　　处：《中国中医基础医学杂志》2023年第11期1847-1850,共4页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然科学基金项目(81201558);河南省科技发展计划项目(142102310263)。

摘　　要：目的 研究川芎嗪(tetramethylpyrazine,TMP)对H22肝癌小鼠血管生成的影响及其分子机制。方法 40只BALB/c小鼠,皮下接种H22肝癌肿瘤细胞悬液建立荷瘤模型后随机分为模型组,低、高剂量组(TMP 25,50 mg/kg),阳性对照组(盐酸多柔比星2 mg/kg),连续给药10 d。ELISA检测各组小鼠血清谷丙转氨酶(glutamic-pyruvic transaminase,ALT)、门冬氨酸转氨酶(glutamic oxalacetic transaminase,AST)水平;计算抑瘤率;苏木精-伊红(hematoxylin eosin,HE)染色观察肿瘤组织的病理形态学变化;免疫组化染色观察肿瘤组织的微血管生成,以CD34标记肿瘤微血管;Western blot检测肿瘤组织中Kruppel样因子4(Kruppellike factor 4,KLF4)、血管内皮生长因子(vascular endothelial growth factor,VEGF)、人金属肽酶含血小板反应蛋白(human A disintegrin and metalloprotease,ADAMTS) 1的表达变化。结果 连续治疗10 d后,与模型组比较,低、高剂量组、阳性对照组小鼠的肿瘤质量和转氨酶ALT、AST水平显著下降(P<0.01),癌巢数量减少,肿瘤细胞出现坏死和空泡样表现,肿瘤组织中KLF4、ADAMTS1蛋白表达显著增高,而VEGF蛋白表达显著降低(P<0.01)。免疫组化染色结果示模型组小鼠肿瘤组织CD34阳性表达率高,可见大量微血管生成,低、高剂量组小鼠随着TMP剂量的增加,肿瘤血管生成逐渐减少。结论 川芎嗪对H22肝癌小鼠的肿瘤生长具有一定的抑制作用,该作用与其调控KLF4、VEGF、ADAMTS1的蛋白表达,进而抑制肿瘤血管生成有关。Objective To investigate the effects of tetramethylpyrazine(TMP)on angiogenesis in H22 hepatoma mice and its molecular mechanism.Methods 40 BALB/c mice were subcutaneously inoculated with H22 hepatocellular carcinoma suspension to establish a tumor-bearing model,then randomly divided into the model group,low dose and high dose group(TMP 25,50 mg/kg),and positive control group(doxorubicin hydrochloride DOX 2 mg/kg).The intervention lasted 10 d.The levels of glutamic-pyruvic transaminase(ALT)serum and glutamic oxaloacetic transaminase(AST)of all groups were detected by ELISA.The inhibitory rate of the tumor was calculated.Hematoxylin-eosin staining was used to observe the pathological changes in tumor tissues.Immunohistochemical staining was used to observe tumor microangiogenesis,and CD34 was used to sign tumor microvessel.Western blotting was used to detect the expression of Kruppel-like factor 4(KLF4),vascular endothelial growth factor(VEGF),and human metallopeptidase platelet reactive protein 1(ADAMTS1).Results After 10 days of continuous treatment,the tumor weight and serum ALT and AST were significantly decreased in low dose&high dose and positive control group compared with the model group(P<0.01).The cancer nests were reduced,the tumor cells were discovered necrosis and vacuole-like.The protein expressions of KLF4 and ADAMTS1 were significantly increased in tumor tissues where VEGF was significantly decreased(P<0.01).Immunohistochemical staining found positive rate of CD34 expression was high and a large number of microangiogenesis were found in the tumor tissues of model group.Angiogenesis gradually decreased along with increasing dosage of TMP in the low-dose and high-dose groups.Conclusion TMP has an inhibitory effect on tumor growth in H22 hepatocellular carcinoma mice,which is related to its inhibitory effect of tumor angiogenesis by means of regulating the protein expressions of KLF4,VEGF,ADAMTS1.

关 键 词：川芎嗪 肝细胞癌 血管生成 VEGF KLF4 ADAMTS1 

分 类 号：R285.5[医药卫生—中药学]