摘要
目的观察SLC5A8基因过表达后,脂多糖诱导的炎症性肠病细胞模型结构及白细胞介素(IL)-8、转化生长因子(TGF)-β_(1)水平变化。方法培养人正常结肠上皮细胞并分为对照组、模型组及实验组。对照组转染pcDNA3.1-GFP空载质粒;模型组转染pcDNA3.1-GFP空载质粒,并以100µg/mL的脂多糖刺激24 h;实验组转染过表达SLC5A8基因质粒,并以100µg/mL的脂多糖刺激24 h。透射电镜下观察三组结肠上皮细胞结构,采用RT-qPCR法检测三组细胞中的IL-8、TGF-β_(1)mRNA,采用ELISA法检测培养液上清中的IL-8、TGF-β_(1)。结果对照组细胞膜表面微绒毛无断裂,无脱落,紧密连接结构致密呈带状;模型组细胞膜表面微绒毛变稀疏,脱落,紧密连接的细胞间隙增宽;实验组细胞膜表面存在微绒毛脱落,紧密连接结构部分破坏。与对照组相比,模型组IL-8 mRNA表达及培养液上清中IL-8水平升高,TGF-β_(1)mRNA表达及培养液上清中TGF-β_(1)水平降低(P均<0.01);与模型组相比,实验组IL-8 mRNA表达及培养液上清中IL-8水平降低,TGF-β_(1)mRNA表达及培养液上清中TGF-β_(1)水平升高(P均<0.01)。结论SLC5A8基因过表达有助于维持炎症性肠病细胞模型的结构,抑制炎症细胞因子IL-8表达,促进抗炎细胞因子TGF-β_(1)表达,从而有助于减轻炎症性肠病的炎症反应。
Objective To observe the changes in morphology of lipopolysaccharide-induced inflammatory bowel disease cell model and levels of interleukin(IL)-8 and transforming growth factor(TGF)-β_(1) after SLC5A8 gene overexpression.Methods Human normal colon epithelial cells were cultured and divided into the control group,model group and experimental group.Cells in the control group were transfected with pcDNA3.1-GFP empty plasmid,cells in the model group were transfected with pcDNA3.1-GFP empty plasmid and then were stimulated with 100μg/mL lipopolysaccharide for 24 h,and the cells in the experimental group were transfected with overexpressed SLC5A8 gene plasmid and stimulated with 100μg/mL lipopolysaccharide for 24 h.The structure of colonic epithelial cells in the three groups was observed under transmission electron microscope.IL-8 and TGF-β_(1) mRNA in cells of the three groups were detected by RT-qPCR,and IL-8 and TGF-β_(1) in the supernatant of culture medium were detected by ELISA.Results The microvilli on the cell membrane surface of the control group were not broken and exfoliated,and the tight junction structure was dense and banded;in the model group,the microvilli on the cell membrane surface became sparse,fell off,and the gap between the tight junction cells widened;there was shedding of microvilli on the cell membrane surface of the experimental group,and the tight junction structure was partially destroyed.Compared with the control group,the expression of IL-8 mRNA and the level of IL-8 in the culture supernatant of the model group increased,while the expression of TGF-β_(1) mRNA and the level of TGF-β_(1) in the culture supernatant decreased(all P<0.01).Compared with the model group,the expression of IL-8 mRNA and the level of IL-8 in the culture supernatant of the experimental group were reduced,while the expression of TGF-β_(1) mRNA and the level of TGF-β_(1) in the culture supernatant increased(all P<0.01).Conclusion Overexpression of SLC5A8 gene helps maintain the morphology of lipopolysaccha
作者
赵泽满
吴育朔
刘雅涵
矫政洧
蔡思源
靳小石
ZHAO Zeman;WU Yushuo;LIU Yahan;JIAO Zhengwei;CAI Siyuan;JIN Xiaoshi(School of Clinical Medicine,Hebei University,Baoding 071000,China;不详)
出处
《山东医药》
CAS
2023年第32期30-33,共4页
Shandong Medical Journal
基金
河北省重点研发计划项目(2137773D)。
