清热化湿方对MNU联合幽门螺杆菌感染相关胃癌细胞中CD14、NF-κB及IL-6表达的影响
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摘要
目的观察清热化湿方对N-甲基-N-亚硝基脲(MNU)联合幽门螺杆菌感染相关胃癌组织中CD14、NF-κB及IL-6表达的影响,探讨清热化湿方防治胃癌可能的机制。方法取90只雄性5周龄BALB/c小鼠,随机分为正常组、模型组、清热化湿方治疗组(低、中、高剂量组)、对照组共6组,每组15只。除正常组外,其余各组采用MNU联合幽门螺杆菌灌胃法制备胃癌模型。幽门螺杆菌定植成功后,低、中、高剂量组分别予低、中、高剂量清热化湿方药液灌胃,对照组予西药灌胃,正常组及模型组予等剂量蒸馏水灌胃。给药至第38周末后取材,采用透射电镜观察小鼠胃黏膜组织病理学变化及超微结构改变;Western blot和Real-time PCR分别检测胃黏膜CD14、NF-κB、IL-6蛋白及mRNA的表达,ELISA测定其表达量。结果模型组镜下见正常黏膜局部脱失,纤维组织增生,炎性细胞浸润,不规则异型腺体增生,呈中分化腺癌改变。治疗组见不同程度黏膜上皮细胞排列不整、缺失,呈散在溃疡改变,少许炎性细胞浸润,未见异型细胞。模型组CD14、NF-κB蛋白及mRNA的表达上升,治疗组表达下降(P<0.05),而各组间IL-6蛋白及mRNA的表达差异无统计学意义(P>0.05);模型组CD14、NF-κB、IL-6的含量表达上升,治疗组表达下降(P<0.05)。结论清热化湿方可能通过下调CD14、NF-κB的表达以达到防治幽门螺杆菌感染相关胃癌的作用。
引文
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[3] Li K,Dan Z,Hu X,et al.CD14 overexpression upregulates TNF-α-mediated inflammatory responses and suppresses the malignancy of gastric carcinoma cells[J].Mol Cell Biochem,2013,376(1-2):137.
[4] Zhou Y,Xia L,Liu Q,et al.Induction of Pro-Inflammatory Response via Activated Macrophage-Mediated NF-κB and STAT3 Pathways in Gastric Cancer Cells[J].Cell Physiol Bioch,2018,47(4):1399.
[5] 许文彬,陈远能,张涛,等.清热化湿方调节PI3K-AKT信号诱导自噬维持胃黏膜稳态防治幽门螺杆菌感染相关胃癌的机制[J].中华中医药学刊,2016,34(10):2335.
[6] 张涛,黄会云,陈思羽,等.稳定沉默CD14胃癌SGC-7901细胞系变化及清热化湿类方药干预研究[J].时珍国医国药,2015,26(3):762.
[7] 孙艳珍,张涛,陈良荣,等.幽门螺杆菌联合MNU灌胃法制备balb/c小鼠胃癌模型研究[J].重庆医学,2017,46(20):2806.
[8] Plummer M,Franceschi S,Vignat J,et al.Global burden of gastric cancer attributable to Helicobacter pylori[J].Int J Cancer,2015,136(2):487.
[9] 赵婕,赵丽华.胃热汤治疗幽门螺杆菌阳性慢性萎缩性胃炎疗效观察[J].河南中医,2018,38(3):431.
[10] 林秋蓉,张怡,陈冠儒,等.邪正发病学说与消化性溃疡发病之关联探析[J].亚太传统医药,2018,14(1):107.
[11] 史娇.半夏泻心汤加味治疗幽门螺杆菌相关性胃炎的效果评定[J].当代医药论丛,2018,16(13):190.
[12] 平倩,张涛,陈玉,等.CD14启动子区多态性介导幽门螺杆菌参与胃癌中上皮-间质转化路径的机制研究[J].广东医学,2014,35(24):3916.
[13] Gong AM,Li XY,Xie YQ,et al.Association between CD14 SNP -159 C/T and gastric cancer:an independent case-control study and an updated meta-analysis[J].Onco Targets Ther,2016,9:4337.
[14] Li K,Dan Z,Hu XJ,et al.Association of CD14/-260 polymorphism with gastric cancer risk in Highland Tibetans[J].World Gastroenterol,2014,20(10):2688.
[15] Mantovani A,Allavena P,Sica A.Cancer-related inflammation[J].Nature,2008,454(7203):436.
[16] Hanahan D.Hallmarks of cancer:the next generation[J].Cell,2011,144(5):646.
[17] Leung TF,Tang NL,Wong GW.CD14 and toll-like receptors:potential contribution of genetic factors and mechanisms to inflammation and allergy[J].Curr Drug Targets Inflamm Allergy,2005,4(2):169.
[18] O'Reilly LA,Putoczki TL,Mielke LA,et al.Loss of NF-κB1 Causes Gastric Cancer with Aberrant Inflammation and Expression of Immune Checkpoint Regulators in a STAT-1-Dependent Manner[J].Immunity,2018,48(3):570.
[19] Li K,Dan Z,Nie Y,et al.CD14 knockdown reduces lipopolysaccharide-induced cell viability and expression of inflammation-associated genes in gastric cancer cells in vitro and in nude mouse xenografts[J].Mol Med Rep,2015,12(3):4332.
[20] Li X,Li M,Huang S,et al.The effect of buffalo CD14 shRNA on the gene expression of TLR4 signal pathway in buffalo monocyte/macrophages[J].Cel Mol Biol Lett,2014,19(4):623.