温肾消水膏皮肤给药舒缓H22肝癌腹水瘤模型小鼠癌性腹水的作用机制研究  

Study on mechanism of skin administration of Wenshen Xiaoshui Ointment in relieving malignant ascites in the murine H22 hepatoma ascites mouse model

作　　者：李阳 张洽淳 陈蕾 纪意纯 钟悦 王济国 冯婧宇 张晶 黄国栋 莫潘艳 LI Yang;ZHANG Qiachun;CHEN Lei;JI Yichun;ZHONG Yue;WANG Jiguo;FENG Jingyu;ZHANG Jing;HUANG Guodong;MO Panyan(Department of Oncology,Bao'an Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Chinese Medicine,Shenzhen,Guangdong 518100;Department of Emergency,Bao'an Hospital of Traditional Chinese MedicineAffiliated to Guangzhou University of Chinese Medicine,Shenzhen,Guangdong 518100;Department of Pathology,Bao'an Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Chinese Medicine,Shenzhen,Guangdong 518100;Department of Central Laboratory,Bao'an Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Chinese Medicine,Shenzhen,Guangdong 518100;Department of Traditional Chinese Medicine,Zigong First People's Hospital,Zigong,Sichuan 643000)

机构地区：[1]广州中医药大学附属宝安中医院肿瘤科,广东深圳518100 [2]广州中医药大学附属宝安中医院急诊科,广东深圳518100 [3]广州中医药大学附属宝安中医院病理科,广东深圳518100 [4]广州中医药大学附属宝安中医院中心实验室,广东深圳518100 [5]四川省自贡市第一人民医院中医科,四川自贡643000

出　　处：《河北中医》2023年第12期2026-2031,共6页Hebei Journal of Traditional Chinese Medicine

基　　金：2020年度广东省中医药局科研项目(编号:20201303);2020年深圳市宝安区医疗卫生基础研究项目(编号:2020JD486);2020年深圳市宝安中医药发展基金会“新冠肺炎治疗和疫情防控技术研究及应用中医药专项”科研项目(编号:2020KJCX-KTYJ-37)。

摘　　要：目的观察温肾消水膏皮肤给药舒缓H22肝癌腹水瘤模型小鼠癌性腹水的作用机制。方法将9只C57BL/6小鼠随机分为模型组、温肾消水膏组、空白组,每组3只,其中模型组、温肾消水膏组采用腹腔接种H22肝癌细胞悬液的方法建立H22肝癌腹水瘤小鼠模型。空白组和模型组不予药物,温肾消水膏组从接瘤后24 h开始,每日予温肾消水膏外敷腹部,每天4 h,连续10天。于第11天以颈椎脱臼法处死小鼠,取腹水、肾脏组织;用注射器计量腹水量,采用显微镜计数法计算肿瘤细胞数,用蛋白质免疫印迹(Western Blot)法检测肾脏组织的水通道蛋白1(AQP1)、水通道蛋白2(AQP2)、血管内皮生长因子A(VEGF-A)和肿瘤坏死因子α(TNF-α)蛋白表达,用逆转录聚合酶链式反应(RT-PCR)法检测肾脏组织中AQP1、AQP2、TNF-α、VEGF-A mRNA表达水平;对肾脏组织进行苏木素-伊红(HE)染色和马松(Masson)染色,观察肾脏的病理形态学改变。结果与模型组比较,温肾消水膏组腹水量减少,但比较差异无统计学意义(P>0.05)。与模型组比较,温肾消水膏组腹水瘤细胞计数减少,但比较差异无统计学意义(P>0.05)。与空白组比较,模型组、温肾消水膏组小鼠肾脏组织VEGF-A蛋白表达均降低(P<0.05),AQP1、AQP2、TNF-α蛋白表达比较差异无统计学意义(P>0.05)。与模型组比较,温肾消水膏组小鼠肾脏组织AQP1蛋白表达升高(P<0.05),AQP2、TNF-α、VEGF-A蛋白表达比较差异无统计学意义(P>0.05)。与空白组比较,模型组、温肾消水膏组肾脏VEGF-A mRNA表达均降低(P<0.05),AQP1、AQP2、TNF-αmRNA表达比较差异无统计学意义(P>0.05)。与模型组比较,温肾消水膏组小鼠肾脏组织AQP1、AQP2、TNF-α、VEGF-A mRNA表达比较差异无统计学意义(P>0.05)。HE染色结果显示,与空白组比较,温肾消水膏组、模型组小鼠肾小球系膜区增宽,系膜细胞增生,肾小管变性,部分区域肾小球萎缩,肾间质有炎性细胞浸润Objective To investigate the mechanism of Wenshen Xiaoshui Ointment in relieving malignant ascites in the murine H22 hepatoma ascites mouse model.Methods Nine C57BL/6 mice were randomly divided into model group,Wenshen Xiaoshui Ointment group and blank group,with 3 mice in each group.The H22 hepatoma ascites mouse model was established by the intraperitoneal inoculation of H22 hepatoma cell suspension in mice of model group and Wenshen Xiaoshui Ointment group.Mice in the blank group and the model group were not intervened by medication.Wenshen Xiaoshui ointment was applied to the abdomen for 4 hours per day at 24 hours of tumor inoculation,for 10 days.On the 11^(th) day,mice were sacrificed by cervical dislocation,and ascites and renal tissues were obtained.The volume of ascites was measured with a syringe,and the number of tumor cells was calculated by microscopic counting method.Protein expressions of aquaporin-1(AQP1),aquaporin-2(AQP2),vascular endothelial growth factor-a(VEGF-A)and tumor necrosis factor-alpha TNF-cein renal tissues were detected by Western Blot.The mRNA expressions of AQP1,AQP2,TNF-by Western B in renal tissues were detected by reverse transcription and polymerase chain reaction polymerase chain reaction(RT-PCR).Renal tissues were subjected to hematoxylin and eosin(H&E)and Masson stainingto observe the pathological morphological changes of the kidneys.Results Compared with that of the model group,the number of ascitestumor cell sandascites volumeinthe Wenshen Xiaoshui Ointment groupdecreased,butthere were no significant differences(P>0.05).Compared with those of the blank group,VEGF-A protein expression was significantly downregulated in the renal tissues of the model group and Wenshen Xiaoshui Ointment group(P<0.05),but there were no significant differences in the protein expressions of AQP1,AQP2 and TNF-α(P>0.05).Compared with those of the model group,AQP1 protein expression was significantly upregulated in the renal tissues of Wenshen Xiaoshui Ointment group(P<0.05),but there were no sign

关 键 词：肝肿瘤 晚期 腹水 小鼠 动物模型 中药外敷 水通道蛋白质1 水通道蛋白质2 

分 类 号：R442.5[医药卫生—诊断学] R244[医药卫生—临床医学]