CAJ | 학술논문

目的：建立小肠隐窝细胞株（IEC-6）药理实验方法。方法：观察细胞接种密度、α-二氟甲基鸟氨酸（DFMO）和胃泌素对IEC-6细胞增殖及细胞鸟氨酸脱羧酶（ODC）的影响。结果：较高接种密度（＞0.5×104细胞/孔）组，接种后第2天，细胞生长抑制，尤其是密度增加到4×104细胞/孔时，第2～3天，OD值逐渐增加，第4天明显增加，第5天达高峰，此后OD值开始下降。而低密度（0.2×104细胞/孔）接种后第4天，细胞生长出现抑制，此后其生长与其他密度相似。加入DFMO后1～3天，完全抑制了细胞增殖，此后与空白组一样，细胞逐渐生长。 DFMO还明显抑制IEC-6细胞分化和移行，以及 ODC活性和腐胺含量，与空白组比较具有显著性差异（P＜0.01）。IEC-6细胞在胃泌素250μg·L-1作用后第一天，开始增殖，第3天达高峰。胃泌素500μg·L-1作用后第1～2天，细胞开始增殖，第3天以后，细胞增殖下降。胃泌素明显促进细胞分化和移行。与空白组比较，胃泌素250μg·L-1分别使ODC mRNA水平，ODC活性，腐胺含量增加1.09倍（P＜0.05），1.71倍（P＜0.01）和5.30倍（P＜0.01）。同样，胃泌素500μg·L-1可使以上3项指标分别升高1.16倍（P＜0.05），1.63倍（P＜0.05）和4.41倍（P＜0.01）。但胃泌素两个剂量组之间无显著性差异。结论：IEC-6细胞是进行胃肠粘膜修复药理实验的合适细胞模型。
목적：건립소장은와세포주（IEC-6）약리실험방법。방법：관찰세포접충밀도、α-이불갑기조안산（DFMO）화위비소대IEC-6세포증식급세포조안산탈최매（ODC）적영향。결과：교고접충밀도（＞0.5×104세포/공）조，접충후제2천，세포생장억제，우기시밀도증가도4×104세포/공시，제2～3천，OD치축점증가，제4천명현증가，제5천체고봉，차후OD치개시하강。이저밀도（0.2×104세포/공）접충후제4천，세포생장출현억제，차후기생장여기타밀도상사。가입DFMO후1～3천，완전억제료세포증식，차후여공백조일양，세포축점생장。 DFMO환명현억제IEC-6세포분화화이행，이급 ODC활성화부알함량，여공백조비교구유현저성차이（P＜0.01）。IEC-6세포재위비소250μg·L-1작용후제일천，개시증식，제3천체고봉。위비소500μg·L-1작용후제1～2천，세포개시증식，제3천이후，세포증식하강。위비소명현촉진세포분화화이행。여공백조비교，위비소250μg·L-1분별사ODC mRNA수평，ODC활성，부알함량증가1.09배（P＜0.05），1.71배（P＜0.01）화5.30배（P＜0.01）。동양，위비소500μg·L-1가사이상3항지표분별승고1.16배（P＜0.05），1.63배（P＜0.05）화4.41배（P＜0.01）。단위비소량개제량조지간무현저성차이。결론：IEC-6세포시진행위장점막수복약리실험적합괄세포모형。
Objective:To established a pharmacological experimental methods of rat small intestinal crypt cell line.Methods:To observed the role of plating densities,alpha-difluoromethylornithine(DFMO)and pentagastrin on the proliferation,differentiation,migration , ODC mRNA level,ODC activity and putrescine content of IEC-6 cells in vitro. Results:Higher plating densities (>0.5×104 cells/well) inhibited the growth of cells on day 2 , especially when a density reach to 4 × 104 cells/well , the OD value increased gradually from day 2 to day 3 , significantly increased on day 4 and reached a peak on day 5 , since then , the OD value begun to reduce . Nevertheless,the growth of IEC-6 was limited at a low density(0.2×104 cells/well) on day 4. After day 4,the growth of the cells was similar with other densities. In addition,DFMO caused a complete inhibition of proliferation of IEC-6 on day 1 to day 3. Since then, the cells grew gradually as well as control group . DFMO also inhibited significantly the differentiation , migration , ODC mRNA level , ODC activity and putrescine content of IEC-6 . Exposure of IEC-6 cells to pentagastrin the proliferation increased initially on d1 and reach a peak on d3 in 250μg·L-1 concentration . Pentagastrin 500μg·L -1 increased cell proliferation on day 1 and day 2 and then decreased.Compared with control group. Pentagastrin stimulated Significantly the differentiation and migration of IEC-6.pentagastrin 250μg·L-1 increased ODC mRNA level 1 . 09-fold(P<0 . 05), ODC activity 1 . 71-fold(P<0 . 01), and putrescine content 5 . 30-fold (P<0.01) respectively. Similarly,pentagastrin 500μg·L-1 also increased ODC mRNA level 1.16-fold (P<0.05),ODC activity 1.63-fold (P<0.05),and putrescine content 4.41-fold (P<0.01) respectively. But there is not significant difference between them. Conclusion:IEC-6 cells are the suitable cell model of gastrointestinal mucosal repair pharmacological experiments .