基于M1型巨噬细胞极化-慢性炎症-肝细胞恶变探究益脾养肝方治疗肝癌前病变作用机制  

Exploration on the Mechanism of Yipi Yanggan Prescription for the Treatment of Liver Precancerous Lesion Based on M1 Type Macrophage Polarization-Chronic Inflammation-Liver Cell Malignant Transformation

作　　者：黄玉 苟雪源 关茜 焦俊喆 闫曙光[1] 常占杰 闫瑞娟 李京涛 HUANG Yu;GOU Xueyuan;GUAN Xi;JIAO Junzhe;YAN Shuguang;CHANG Zhanjie;YAN Ruijuan;LI Jingtao(The First Clinical Medical School of Shaanxi University of Chinese Medicine,Xianyang 712000,China;The Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,China)

机构地区：[1]陕西中医药大学第一临床医学院,陕西咸阳712000 [2]陕西中医药大学附属医院,陕西咸阳712000

出　　处：《中国中医药信息杂志》2024年第10期81-88,共8页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82174330、82374418);陕西省重点研发计划(2020ZDLSF05-15);陕西省自然科学基础研究计划(2022JQ-965);陕西省创新能力支撑计划(2022TD-55);陕西省中医药管理局委托项目(SZY-KJCYC-2023-049、SZY-KJCYC-2023-087);陕西省中医药管理局中医药“双链融合”中青年科研创新团队项目(2022-SLRH-LJ-002);咸阳市重点研发计划(L2022ZDYFSF007);咸阳市重大科技创新专项(L2023-ZDKJ-CYJQ-SF-011);国家中医药管理局全国名老中医药专家传承工作室建设项目(2022年)。

摘　　要：目的基于M1型巨噬细胞极化-慢性炎症-肝细胞恶变发展过程探究益脾养肝方治疗肝癌前病变大鼠的作用机制。方法90只Wistar大鼠随机分为空白组、模型组、护肝片组和益脾养肝方高、中、低剂量组,每组15只。空白组予蒸馏水腹腔注射,其余各组予二乙基亚硝胺腹腔注射诱导肝癌前病变模型,每周2次(50 mg/kg),连续16周。造模第2日起,益脾养肝方高、中、低剂量组分别予1.2、0.6、0.3 g/mL益脾养肝方灌胃,护肝片组予护肝片溶液921 mg/kg灌胃,空白组和模型组予等量生理盐水灌胃,连续16周。观察肝脏外观,ELISA检测血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转氨酶(AST)、碱性磷酸酶(ALP)、α-L-岩藻糖苷酶(AFU)及肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、诱导型一氧化氮合酶(iNOS)、单核细胞趋化蛋白-1(MCP-1)含量,HE染色、Masson染色观察肝组织形态,免疫组化检测肝细胞恶变标志物OV6、细胞角蛋白19(CK19)、CD133和上皮细胞黏附分子(EpCAM)表达,qPCR检测肝组织CK19、CD133、EpCAM mRNA表达,免疫荧光共定位检测M1型巨噬细胞标志物CD68与IL-6和TNF-α共表达。结果与空白组比较,模型组大鼠肝脏质硬,表面粗糙、边缘变钝,可见大量结节,血清ALT、AST、ALP、AFU、TNF-α、IL-6、i NOS、MCP-1含量明显升高(P<0.01),肝组织有大面积异型增生结节、炎性细胞浸润,胶原纤维增多,肝组织OV6、CK19、CD133、EpCAM表达显著升高,CD68与IL-6和TNF-α共表达显著升高(P<0.01);与模型组比较,各给药组大鼠肝脏结节减少且较小,血清ALT、AST、ALP、AFU、TNF-α、IL-6、iNOS、MCP-1含量均显著降低(P<0.01),肝细胞异型增生及炎性细胞浸润显著改善,胶原纤维减少,肝组织OV6、CK19、CD133、EpCAM表达显著降低,CD68与IL-6和TNF-α共表达显著降低(P<0.05,P<0.01)。结论益脾养肝方可能通过抑制M1型巨噬细胞极化减轻炎症反应,改善肝细胞恶变,对二乙基亚硝Objective To explore the mechanism of Yipi Yanggan Prescription in the treatment of precancerous lesion of liver in rats based on M1 type macrophage polarization-chronic inflammation-liver cell malignant transformation.Methods Totally 90 Wistar rats were randomly divided into blank group,model group,Hugan Tablet group and Yipi Yanggan Prescription high-,medium-and low-dosage groups,with 15 rats in each group.The blank group was injected distilled water intraperitoneally,and the other groups were injected 5 mL/kg of diethylnitrosamine intraperitoneally at 50 mg/kg per week(twice per week)for 16 weeks to induce the precancerous lesion of liver model.Starting from the second day of modeling,Yipi Yanggan Prescription high-,medium-and low-dosage groups were orally administered with 1.2,0.6 and 0.3 g/mL Yipi Yanggan Prescription,respectively.The Hugan Tablet group was orally administered with 921 mg/kg Hugan Tablet solution,the blank group and model group were orally administered with an equal amount of physiological saline for 16 consecutive weeks.The appearance of the liver was observed,ELISA was used to detect serum ALT,AST,ALP,AFU,as well as TNF-α,IL-6,iNOS and MCP-1 content,HE staining and Masson staining were used to observe the morphology of liver tissue,immunohistochemistry was used to detect the expressions of liver cell malignancy markers OV6,CK19,CD133 and EpCAM,qPCR was used to detect the mRNA expressions of CK19,CD133 and EpCAM in liver tissue,immunofluorescence co-localization was used to detect the co-expressions of M1 type macrophage markers CD68 with IL-6 and TNF-α.Results Compared with the blank group,the liver of the model group rats was hard,with a rough surface and dull edges,and a large number of nodules were visible,the contents of serum ALT,AST,ALP,AFU,TNF-α,IL-6,iNOS and MCP-1 significantly increased(P<0.01),there were large areas of dysplasia nodules,inflammatory cell infiltration,and increased collagen fibers in liver tissue,the expressions of OV6,CK19,CD133 and EpCAM in liver tissue signi

关 键 词：益脾养肝方 肝癌前病变 肝细胞恶变 慢性炎症 M1型巨噬细胞 大鼠 

分 类 号：R285.5[医药卫生—中药学]