基于HIF-1α/VEGF信号通路探讨大黄䗪虫丸抑制KM小鼠肿瘤血管的机制研究  

The Mechanism of Dahuang Zhechong Pill Inhibiting Tumor Angiogenesis in KM Mice Based on the HIF-1α/VEGF Signaling Pathway

作　　者：李盼丽 李钦青 王光琦 柳甜 郭继龙 贺文彬 LI Pan-li;LI Qin-qing;WANG Guang-qi;LIU Tian;GUO Ji-long;HE Wen-bin(National International Joint Research Center for Molecular Chinese Medicine,Shanxi University of Chinese Medicine,Taiyuan Shanxi 030024;Shanxi Key Laboratory of Chinese Medicine Encephalopathy,Shanxi University of Chinese Medicine,Taiyuan Shanxi 030024;The Fourth Clinical College,Shanxi University of Chinese Medicine,Taiyuan Shanxi 030024)

机构地区：[1]山西中医药大学分子中医药学国家级国际联合研究中心,山西太原030024 [2]山西中医药大学,中医脑病学山西省重点实验室,山西太原030024 [3]山西中医药大学第四临床学院,山西太原030024

出　　处：《世界中西医结合杂志》2024年第8期1559-1565,共7页World Journal of Integrated Traditional and Western Medicine

基　　金：山西省基础研究计划项目(20090311053)。

摘　　要：目的 研究大黄䗪虫丸对昆明(KM)小鼠移植瘤的抑瘤率、体积、质量、缺氧诱导因子(Hypoxia inducible factor-1,HIF-1α)、血管内皮生长因子(Vascular endothelial growth factor, VEGF)、肿瘤血管密度(Microvascular density, MVD)的影响,探讨大黄䗪虫丸是否是通过HIF-1α/VEGF信号通路达到抑制肿瘤血管生成进而控制肿瘤生长的作用。方法 将H22人肝癌细胞注射到KM小鼠进行小鼠皮下移植瘤造模,造模成功后按照随机数字表法分为空白组、模型组、大黄䗪虫丸低剂量组、大黄䗪虫丸中剂量组、大黄䗪虫丸高剂量组和索拉非尼组,每组各15只,实验干预1次/d,连续干预2周。大黄䗪虫丸低剂量组、中剂量组、高剂量组和索拉非尼组均给予相应药物灌胃,模型组给予生理盐水灌胃。实验干预结束后取瘤体,对各组瘤体长、短径进行测量,计算瘤体的抑瘤率、体积及体质量并记录;采用苏木精-伊红染色法检测各组肿瘤组织形态学的病理变化;实时荧光定量聚合酶链反应法检测HIF-1α和VEGF mRNA转录水平;蛋白印记法检测HIF-1α和VEGF蛋白表达水平;采用免疫组织化法检测肿瘤组织检测反映MVD的表达。结果 体积抑瘤率比较,大黄䗪虫丸低、中、高剂量组的抑瘤率分别为17.11%、21.73%和37.09%,索拉非尼组的抑瘤率为46.75%;HE染色结果显示:与模型组比较,大黄䗪虫丸高剂量组可使肿瘤组织出现大面积的坏死区域;PCR和WB结果显示:与模型组比较,大黄䗪虫丸各治疗组均可不同程度的显著降低肿瘤组织HIF-1α、VEGF mRNA蛋白水平表达(P<0.05,P<0.01);CD31-MVD结果显示:与模型组比较,大黄䗪虫丸高剂量组MVD明显减少(P<0.05)。结论 大黄䗪虫丸可能是通过HIF-1α/VEGF信号通路达到抑制肿瘤血管生成进而控制肿瘤的生长。Objective To investigate the mechanism by which the Dahuang Zhechong pill inhibits tumor angiogenesis in Kunming(KM)mice,focusing on its effects on tumor inhibition rate,tumor volume,tumor mass,hypoxia-inducible factor-1α(HIF-1α),vascular endothelial growth factor(VEGF),and tumor microvascular density(MVD).The study aims to explore whether the Dahuang Zhechong pill inhibits tumor angiogenesis and control tumor growth via the HIF-1α/VEGF signaling pathway.Methods H22 human liver cancer cells were injected into KM mice to establish subcutaneous tumor models.After successful modeling,the mice were randomly divided into six groups:a blank group,a model group,a low-dose Dahuang Zhechong pill group,a medium-dose group,a high-dose group,and a Sorafenib group,with 15 mice in each group.The mice were administered the experimental interventions once daily for two consecutive weeks.The Dahuang Zhechong pill groups and the Sorafenib group were administered corresponding drugs via gavage,while the model group received saline via gavage.After the intervention,the tumors were excised,and their length and width were measured to calculate tumor inhibition rate,tumor volume,and tumor mass,which were recorded.Hematoxylin-eosin(HE)staining was used to observe pathological changes in tumor tissues.Real-time fluorescent quantitative polymerase chain reaction(qPCR)was used to detect the transcription levels of HIF-1α and VEGF mRNA.Western blot(WB)was used to detect the protein expression levels of HIF-1α and VEGF.Immunohistochemistry was employed to detect MVD in tumor tissues.Results The tumor inhibition rates in the low,medium,and high-dose Dahuang Zhechong pill groups were 17.11%,21.73%,and 37.09%,respectively,while the inhibition rate in the Sorafenib group was 46.75%.HE staining showed that the high-dose Dahuang Zhechong pill group led to large areas of necrosis in tumor tissues compared to the model group.qPCR and WB results indicated that all treatment groups significantly reduced the expression levels of HIF-1α and VEGF m

关 键 词：大黄䗪虫丸 原发性肝癌 H22细胞 缺氧诱导因子 血管内皮生长因子 

分 类 号：R285.5[医药卫生—中药学]