中华小儿外科杂志
中華小兒外科雜誌
중화소인외과잡지
CHINESE JOURNAL OF PEDIATRIC SURGERY
2013年
6期
458-462
,共5页
黄晓忠%林正秀%张田丰%朱利斌%李仲荣%林锦
黃曉忠%林正秀%張田豐%硃利斌%李仲榮%林錦
황효충%림정수%장전봉%주리빈%리중영%림금
脂肪酸类,挥发性%肠黏膜%丝裂原激活蛋白激酶类
脂肪痠類,揮髮性%腸黏膜%絲裂原激活蛋白激酶類
지방산류,휘발성%장점막%사렬원격활단백격매류
Fatty acids,volatile%Intestinal mucosa%Mitogen-activated protein kinases
目的 观察不同浓度丁酸钠对Caco-2细胞肠屏障模型的作用情况及p38MAPK在这一过程中可能的影响,为进一步纠正肠屏障功能异常寻找新靶点.方法 通过Caco-2细胞培养,建立单层Caco-2细胞肠屏障模型；比较对照组、不同浓度丁酸钠(2 mmol/L、5 mmol/L和8 mmol/L)及丁酸钠(5mmol/L)+ SB203580(p38MAPK特异性抑制剂,20 μmol/L)组对Caco-2细胞肠屏障模型的作用情况,测定连续培养72 h后肠屏障模型的跨上皮电阻(TER)、菊粉-FITC(inulin-FITC)的通透性及扫描电镜表现.结果 体外培养的Caco-2细胞能在3周后形成肠上皮细胞屏障模型.低浓度丁酸钠(2 mmol/L)作用于肠屏障模型72 h后,TER达到(467.65±7.41)Ω·cm2,与对照组的(417.58±6.29)Ω·cm2相比差异有统计学意义(P＜0.05)；中、高浓度丁酸钠(5 mmol/L和8mmol/L)作用72 h后,TER分别为(113.87±20.27)Ω·cm2和(58.60±29.02)Ω·cm2,菊粉通透性分别为(11.08±1.04)％和(16.01±1.56)％,与对照组相比差异均有统计学意义(P＜0.05).而5 mmol/L丁酸钠+20 μmol/L SB203580组TER和通透性分别为(395.17±9.97)Ω·cm2和(7.19±0.97)％,与单纯5 mmol/L丁酸钠组相比,差异有统计学意义(P＜0.05).扫描电镜发现对照组和2 mmol/L丁酸钠组单层Caco-2细胞屏障表面完整,细胞连接紧密无脱落；5 mmol/L和8 mmol/L丁酸钠组随药物浓度增加,细胞变形呈球形脱落亦增加；5 mmol/L丁酸钠+20 μmol/L SB203580组细胞脱落较5mmol/L丁酸钠组明显减轻.结论 丁酸钠对Caco-2细胞肠屏障模型作用具有双向性:低浓度丁酸钠促进肠屏障功能,而中、高浓度丁酸钠可破坏肠屏障功能；p38MAPK特异性抑制剂SB203580可减轻中等浓度丁酸钠对肠屏障功能的损害.
目的 觀察不同濃度丁痠鈉對Caco-2細胞腸屏障模型的作用情況及p38MAPK在這一過程中可能的影響,為進一步糾正腸屏障功能異常尋找新靶點.方法 通過Caco-2細胞培養,建立單層Caco-2細胞腸屏障模型；比較對照組、不同濃度丁痠鈉(2 mmol/L、5 mmol/L和8 mmol/L)及丁痠鈉(5mmol/L)+ SB203580(p38MAPK特異性抑製劑,20 μmol/L)組對Caco-2細胞腸屏障模型的作用情況,測定連續培養72 h後腸屏障模型的跨上皮電阻(TER)、菊粉-FITC(inulin-FITC)的通透性及掃描電鏡錶現.結果 體外培養的Caco-2細胞能在3週後形成腸上皮細胞屏障模型.低濃度丁痠鈉(2 mmol/L)作用于腸屏障模型72 h後,TER達到(467.65±7.41)Ω·cm2,與對照組的(417.58±6.29)Ω·cm2相比差異有統計學意義(P＜0.05)；中、高濃度丁痠鈉(5 mmol/L和8mmol/L)作用72 h後,TER分彆為(113.87±20.27)Ω·cm2和(58.60±29.02)Ω·cm2,菊粉通透性分彆為(11.08±1.04)％和(16.01±1.56)％,與對照組相比差異均有統計學意義(P＜0.05).而5 mmol/L丁痠鈉+20 μmol/L SB203580組TER和通透性分彆為(395.17±9.97)Ω·cm2和(7.19±0.97)％,與單純5 mmol/L丁痠鈉組相比,差異有統計學意義(P＜0.05).掃描電鏡髮現對照組和2 mmol/L丁痠鈉組單層Caco-2細胞屏障錶麵完整,細胞連接緊密無脫落；5 mmol/L和8 mmol/L丁痠鈉組隨藥物濃度增加,細胞變形呈毬形脫落亦增加；5 mmol/L丁痠鈉+20 μmol/L SB203580組細胞脫落較5mmol/L丁痠鈉組明顯減輕.結論 丁痠鈉對Caco-2細胞腸屏障模型作用具有雙嚮性:低濃度丁痠鈉促進腸屏障功能,而中、高濃度丁痠鈉可破壞腸屏障功能；p38MAPK特異性抑製劑SB203580可減輕中等濃度丁痠鈉對腸屏障功能的損害.
목적 관찰불동농도정산납대Caco-2세포장병장모형적작용정황급p38MAPK재저일과정중가능적영향,위진일보규정장병장공능이상심조신파점.방법 통과Caco-2세포배양,건립단층Caco-2세포장병장모형；비교대조조、불동농도정산납(2 mmol/L、5 mmol/L화8 mmol/L)급정산납(5mmol/L)+ SB203580(p38MAPK특이성억제제,20 μmol/L)조대Caco-2세포장병장모형적작용정황,측정련속배양72 h후장병장모형적과상피전조(TER)、국분-FITC(inulin-FITC)적통투성급소묘전경표현.결과 체외배양적Caco-2세포능재3주후형성장상피세포병장모형.저농도정산납(2 mmol/L)작용우장병장모형72 h후,TER체도(467.65±7.41)Ω·cm2,여대조조적(417.58±6.29)Ω·cm2상비차이유통계학의의(P＜0.05)；중、고농도정산납(5 mmol/L화8mmol/L)작용72 h후,TER분별위(113.87±20.27)Ω·cm2화(58.60±29.02)Ω·cm2,국분통투성분별위(11.08±1.04)％화(16.01±1.56)％,여대조조상비차이균유통계학의의(P＜0.05).이5 mmol/L정산납+20 μmol/L SB203580조TER화통투성분별위(395.17±9.97)Ω·cm2화(7.19±0.97)％,여단순5 mmol/L정산납조상비,차이유통계학의의(P＜0.05).소묘전경발현대조조화2 mmol/L정산납조단층Caco-2세포병장표면완정,세포련접긴밀무탈락；5 mmol/L화8 mmol/L정산납조수약물농도증가,세포변형정구형탈락역증가；5 mmol/L정산납+20 μmol/L SB203580조세포탈락교5mmol/L정산납조명현감경.결론 정산납대Caco-2세포장병장모형작용구유쌍향성:저농도정산납촉진장병장공능,이중、고농도정산납가파배장병장공능；p38MAPK특이성억제제SB203580가감경중등농도정산납대장병장공능적손해.
Objective To explore the effects of different concentrations of sodium butyrate on a Caco-2 cell monolayer model of intestinal barrier and to investigate the potential influence of p38 MAPK on this process.Methods The Caco-2 cell monolayer model of intestinal barrier was established via cell culture,which was further divided into 3 groups including the control group,the sodium butyrate groups with different concentrations (2 mmol/L,5 mmol/L,and 8 mmol/L)) and the sodium butyrate (5 mmol/L) + SB203580(20μmol/L,p38MAPK specific inhibitor)group.After 72 hours of culture,the transepithelial electrical resistance (TER) and the barrier permeability by testing the concentration of inulin-FITC were detected.The surface of Caco-2 cells was observed by electronic scanning microscopy.Results After 3 weeks of culture,monolayer Caco-2 cells formed the intestinal epithelial cell barrier model.After incubation of sodium butyrate (2 mmol/L) on Caco-2 cell monolayers for 72 h,the TER was increased to (467.65 ± 7.41) Ω·cm2 compared to (417.58 ± 6.29)Ω·cm2 in the control group (P＜0.05).After incubation of sodium butyrate (5 mmol/L or 8 mmol/L) on Caco-2 cells monolayer for 72 h,the TERs were (113.87 ± 20.27)Ω· cm2 and (58.60 ± 29.02)Ω· cm2,which were both significantly lower than those in the control group (P＜0.05).The inulin-FITC permeability was (11.08± 1.04)％ and (16.01 ± 1.56)％ in these 2 groups (P＜0.05,compared to the control group).However,the TER and permeability of the 5 mmol/L sodium butyrate + 20 μmol/L SB203580 group were (395.17 ± 9.97)Ω·cm2 and (7.19 ± 0.97)％ (both P＜0.05,compared to those in the 5 mmol/L sodium butyrate group).The electronic microscopy revealed that the surface of the Caco-2 cells,which were treated with 0 mmol/L and 2 mmol/L sodium butyrate,was connected tightly without cell float.The surface impairment and cell float of Caco-2 cell monolayers were increased in a concentration-dependent fashion.Compared to that in the 5 mmol/L sodium butyrate group,the cell loss in the 5 mmol/L sodium butyrate + 20 μmol/L SB203580 group were significantly decreased.Conclusions The low concentration of sodium butyrate may be beneficial in promoting intestinal barrier functions,whereas excessive sodium butyrate may disrupt normal intestinal barrier.And p38MAPK specific inhibitor SB203580 could decrease the damage of the intestinal barrier function induced by sodium butyrate.
