肠复方对肠癌肿瘤相关巨噬细胞及趋化因子2、白介素10表达的影响  

Experimental Study on the Expression of TAM and CCL2,IL-10 in Colorectal Cancer by Intestinal Compound

作　　者：崔超宇 梁慧 章慧 邓湘琴 林宏远 龙梦铃 谭骏岚 李云辉[3] CUI Chaoyu;LIANG Hui;ZHANG Hui;DENG Xiangqin;LIN Hongyuan;LONG Mengling;TAN Junlan;LI Yunhui(Hunan Chinese Medical University,Changsha 410208 Hunan,China;Hunan Cancer Hospital,Changsha 410013 Hunan,China;Xiangya Hospital,Changsha 410008 Hunan,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]中南大学湘雅医学院附属肿瘤医院,湖南长沙410013 [3]中南大学湘雅医院,湖南长沙410008

出　　处：《中药新药与临床药理》2020年第9期1021-1028,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：湖南省自然科学基金面上项目(2017JJ2175);湖南省中医药管理局重点项目(201807);长沙市科技局项目(kp1701044)。

摘　　要：目的研究肠复方对裸鼠肠癌原位移植瘤不同部位肿瘤相关巨噬细胞(tumor-associated macrophages,TAM)及相关因子的影响。方法复制裸鼠肠癌原位移植瘤模型,将模型复制成功的28只裸鼠随机分为肠复方高(72 g·kg^-1)、中(36 g·kg^-1)、低(18 g·kg^-1)剂量组和蒸馏水组,每组7只。模型复制成功次日为第1天,于第14天开始给予相应药物干预,共干预3周。干预结束后处死裸鼠,剥离肿瘤标本,称质量计算抑瘤率;采用HE染色观察各组肠癌肿块不同部位标本细胞学形态;流式细胞学法检测各组标本M1型、M2型TAM含量;Western Blot及RT-PCR法检测各组标本中趋化因子2(C-C motif ligand 2,CCL2)及白介素10(Interleukin 10,IL-10)的蛋白及mRNA表达水平。结果与蒸馏水组比较,肠复方药物组肿瘤质量明显降低,差异有统计学意义(P<0.05),肠复方高、中、低剂量组抑瘤率分别为36.01%、20.01%、18.00%。无药物干预的蒸馏水组中,M1型、M2型肿瘤相关巨噬细胞(TAM)含量分布趋势一致:癌中心>癌浸润前缘>癌旁组织,与癌旁组织比,差异均具有统计学意义(P<0.01)。与蒸馏水组相应部位比,肠复方高、中、低剂量组癌浸润前缘及癌中心M1型TAM含量均下降,差异有统计学意义(P<0.01,P<0.05);肠复方高、中剂量组M2型TAM含量明显下降,差异有统计学意义(P<0.01),肠复方低剂量组M2型TAM虽有下降趋势,差异无统计学意义(P>0.05)。无药物干预的蒸馏水组中:CCL2、IL-10蛋白及mRNA表达一致:癌中心>癌浸润前缘>癌旁组织,与癌旁组织比差异均有统计学意义(P<0.01)。与蒸馏水组相应部位比较,肠复方各组癌中心CCL2、IL-10蛋白及mRNA表达均明显下降,差异有统计学意义(P<0.01);肠复方各组癌浸润前缘CCL2、IL-10蛋白表达均明显下降,差异有统计学意义(P<0.01);肠复方各组癌浸润前缘CCL2 mRNA(P<0.01)、IL-10 mRNA(P<0.05,P<0.01)表达均有所下降。结论癌组织(癌中心、癌浸润前缘)�Objective To investigate the effect of Intestinal compound on tumor-associated macrophages(TAM) and related factors in different parts of nude mice colon cancer orthotopic transplantation tumor.Methods An orthotopic tumor transplantation model was established in nude mice.Twenty-eight nude mice were randomly divided into high(72 g·kg^-1),medium(36 g·kg^-1),low(18 g·kg^-1) dose groups and distilled water group.Corresponding intervention was started on the 14 th day after the modelling,and the intervention last for 21 days.After the intervention,nude mice were sacrificed and the tumor samples were stripped;the tumor suppression rate was calculated by weighing;the cytology morphology of different parts of intestinal cancer masses in each group was observed by HE staining,and the contents of M1 and M2 TAM in each group were detected by flow cytometry;the protein and mRNA expression levels of chemokine 2(CCL2) and interleukin 10(IL-10) in each group were detected by Western Blot and RT-PCR.Results Compared with the distilled water group,the mass of tumor in the Intestinal compound drug group was significantly reduced(P<0.05).The tumor inhibition rates of the Intestinal compound high,medium and low dose groups were 36.01%,20.01%,18.00%.The contents of M1 and M2(TAM) in the distilled water group without drug intervention were the same:cancer center tissue>cancer invasion front> adjacent tissues,the difference between the first two and the adjacent tissue were statistically significant(P<0.01).The contents of M1 TAM in high,medium and low dose groups of Intestinal compound decreased,and the difference compared with those in distilled water group were statistically significant(P<0.01,P<0.01,P<0.05).The contents of M2 type TAM in high and medium dose groups of Intestinal compound decreased significantly(P<0.01);and there was a downward trend in low dose group of Intestinal compound with no significant difference(P<0.05).In the distilled water group without drug intervention:CCL2,IL-10 protein and mRNA expression were co

关 键 词：肠复方 大肠癌 肿瘤相关巨噬细胞 趋化因子2 白细胞介素10 裸鼠 

分 类 号：R285.5[医药卫生—中药学]