温郁金二萜类化合物C通过miR-223-3p/NF-kB通路对幽门螺杆菌诱导的GES-1细胞的保护作用  

Protective effect of Curcuma wenyujin diterpenoid compound C on GES-1 cells induced by Helicobacter pylori through miR-223-3p/NF-kB pathway

作　　者：刘治智 黄美君[1] 杨颖强[1] 陈伟萍[1] LIU Zhi-zhi;HUANG Mei-jun;YANG Ying-qiang;CHEN Wei-ping(Department of Gastroenterology,Taizhou Hospital,Taizhou Enze Medical Group Enze Hospital,Zhejiang 318050,China)

机构地区：[1]浙江省台州医院、台州恩泽医疗中心(集团)恩泽医院消化内科,浙江318050

出　　处：《中国卫生检验杂志》2023年第7期788-792,共5页Chinese Journal of Health Laboratory Technology

基　　金：台州市科技计划项目(1801KY71)。

摘　　要：目的探索温郁金二萜类化合物C对幽门螺杆菌(Hp)诱导的GES-1细胞的保护作用及机制。方法选取GES-1细胞(空白组),采用Hp感染(模型组),使用不同浓度温郁金二萜类化合物C(10μg/ml、20μg/ml及80μg/ml)处理细胞(低剂量组、中剂量组及高剂量组)。CCK-8法检测细胞增殖,流式细胞术检测细胞凋亡,Transwell检测细胞侵袭、迁移,实时荧光定量PCR(qRT-PCR)检测上皮细胞钙黏蛋白(E-cadherin)、神经钙黏素(N-cadherin)、波形纤维蛋白(Vimentin)基因mRNA及miR-223-3p水平,Western blot检测细胞E-cadherin、N-cadherin、Vimentin、p-p65、p65及核因子κB抑制蛋白α(IKBα)蛋白水平。结果与空白组比较,模型组24 h、48 h及72 h细胞A值升高,凋亡率降低,侵袭、迁移细胞数升高(P<0.05);与模型组比较,温郁金二萜类化合物C处理组24 h、48 h及72 h细胞A值降低,凋亡率升高,侵袭、迁移细胞数降低,且呈现浓度依赖性(P<0.05)。Hp诱导GES-1细胞发生上皮间质转化(EMT),温郁金二萜类化合物C可逆转EMT,且呈现浓度依赖性。与空白组比较,模型组细胞miR-223-3p、p-p65、p65及IKBα蛋白表达水平升高(P<0.05),与模型组比较,温郁金二萜类化合物C处理组细胞miR-223-3p、p-p65、p65及IKBα蛋白表达水平下降,且呈现浓度依赖性(P<0.05)。结论温郁金二萜类化合物C可通过miR-223-3p/NF-κB通路抑制Hp感染GES-1细胞的恶性生物学行为。Objective This paper aims to explore the protective effect and mechanism of Curcuma wenyujin diterpenoid compound C on GES-1 cells induced by Helicobacter pylori(Hp).Methods GES-1 cells(blank group)were infected with Hp(model group),and different concentrations of Curcuma wenyujin diterpenoids compound C(10μg/ml,20μg/ml and 80μg/ml)were used to treat cells(low dose group,medium dose group and high dose group).CCK-8 method was used to detect cell proliferation.Flow cytometry was used to detect cell apoptosis.Transwell was used to detect cell invasion and migration.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the mRNA of E-cadherin,N-cadherin,Vimentin gene and the level of miR-223-3p in epithelial cells.Western blot was used to detect protein level of E-cadherin,N-cadherin,Vimentin,p-p65,p65 and nuclear factorκB inhibitory proteinα(IKBα).Results Compared with the blank group,the A value of cells in the model group increased at 24 h,48 h and 72 h,the apoptosis rate increased,and the number of invasive and migratory cells increased,showing concentration dependence(P<0.05).Compared with the model group,the A value of cells in Curcuma wenyujin diterpenoid compound C treatment group decreased at 24 h,48 h and 72 h,the apoptosis rate increased,and the number of invasive and migratory cells decreased in a concentration dependent manner(P<0.05).Hp induced epithelial mesenchymal transformation(EMT)in GES-1 cells,and Curcuma wenyujin diterpenoid compound C could reverse EMT in a concentration dependent manner.Compared with the blank group,the level of miR-223-3p and the protein levels of p-p65,p65 and IKBαin the model group cells increased(P<0.05).Compared with the model group,the levels of miR-223-3p,p-p65,p65 and IKBαof cells in Curcuma wenyujin diterpene compound C treatment group decreased in a concentration dependent manner(P<0.05).Conclusion Curcuma wenyujin diterpenoid compound C can inhibit the malignant biological behavior of GES-1 cells infected by Hp via miR-223-3p/NF-κB pathway.

关 键 词：温郁金二萜类化合物C miR-223-3p NF-kB通路 幽门螺杆菌 人正常胃黏膜上皮细胞 

分 类 号：R446.5[医药卫生—诊断学]