中华生物医学工程杂志
中華生物醫學工程雜誌
중화생물의학공정잡지
Chinese Journal of Biomedical Engineering
2015年
4期
325-329
,共5页
杨院平%罗和生%仝巧云%李中艳%郑世华
楊院平%囉和生%仝巧雲%李中豔%鄭世華
양원평%라화생%동교운%리중염%정세화
炎症%肠上皮细胞%脂多糖%壳寡糖%TLR4
炎癥%腸上皮細胞%脂多糖%殼寡糖%TLR4
염증%장상피세포%지다당%각과당%TLR4
Inflammation%Intestinal epithelial cells%Lipopolysaccharide%Oligochitosan%TLR4
目的：研究壳寡糖对脂多糖（LPS）诱导肠上皮细胞炎症的影响。方法肠上皮细胞Caco?2分为5组：正常对照组、模型组、壳寡糖高、中、低浓度组。以1μg/L LPS刺激细胞48h建立炎症模型。药物干预组于LPS刺激前2 h，分别以0.25，0.5和1.0 g/L壳寡糖预处理细胞。MTT法检测细胞活力。以ELISA法检测培养上清中炎症相关因子肿瘤坏死因子（TNF）?α，白介素（IL）?8和前列腺素（PG）E2水平。以Western印迹法检测细胞Toll样受体（TLR）4、核因子κB（NF?κB）及环氧酶（COX）2的表达变化。结果壳寡糖和/或LPS处理后Caco?2细胞存活率不受影响。与模型组相比，壳寡糖在0.25，0.5和1.0 g/L剂量下呈浓度依赖性地降低Caco?2细胞TNF?α（352.5±21.6，298.4±25.1，203.4±20.0 vs 436.8±38.7μg/L）和PGE2的释放（632.2±35.6，522.6±26.7，402.4±30.2 vs 822.3±23.5μg/L）（P<0.05；P<0.01），0.5和1.0 g/L壳寡糖可降低细胞COX?2表达水平（P<0.05；P<0.01）。壳寡糖中、高浓度组TLR4表达明显低于模型组（P<0.05），壳寡糖各剂量组NF?κB的表达水平显著降低（P<0.05；P<0.01）。结论壳寡糖可对抗LPS诱导的肠上皮细胞炎症，对炎性肠病具潜在保护作用。作用机理可能与TLR4\NF?κB信号通路抑制有关。
目的：研究殼寡糖對脂多糖（LPS）誘導腸上皮細胞炎癥的影響。方法腸上皮細胞Caco?2分為5組：正常對照組、模型組、殼寡糖高、中、低濃度組。以1μg/L LPS刺激細胞48h建立炎癥模型。藥物榦預組于LPS刺激前2 h，分彆以0.25，0.5和1.0 g/L殼寡糖預處理細胞。MTT法檢測細胞活力。以ELISA法檢測培養上清中炎癥相關因子腫瘤壞死因子（TNF）?α，白介素（IL）?8和前列腺素（PG）E2水平。以Western印跡法檢測細胞Toll樣受體（TLR）4、覈因子κB（NF?κB）及環氧酶（COX）2的錶達變化。結果殼寡糖和/或LPS處理後Caco?2細胞存活率不受影響。與模型組相比，殼寡糖在0.25，0.5和1.0 g/L劑量下呈濃度依賴性地降低Caco?2細胞TNF?α（352.5±21.6，298.4±25.1，203.4±20.0 vs 436.8±38.7μg/L）和PGE2的釋放（632.2±35.6，522.6±26.7，402.4±30.2 vs 822.3±23.5μg/L）（P<0.05；P<0.01），0.5和1.0 g/L殼寡糖可降低細胞COX?2錶達水平（P<0.05；P<0.01）。殼寡糖中、高濃度組TLR4錶達明顯低于模型組（P<0.05），殼寡糖各劑量組NF?κB的錶達水平顯著降低（P<0.05；P<0.01）。結論殼寡糖可對抗LPS誘導的腸上皮細胞炎癥，對炎性腸病具潛在保護作用。作用機理可能與TLR4\NF?κB信號通路抑製有關。
목적：연구각과당대지다당（LPS）유도장상피세포염증적영향。방법장상피세포Caco?2분위5조：정상대조조、모형조、각과당고、중、저농도조。이1μg/L LPS자격세포48h건립염증모형。약물간예조우LPS자격전2 h，분별이0.25，0.5화1.0 g/L각과당예처리세포。MTT법검측세포활력。이ELISA법검측배양상청중염증상관인자종류배사인자（TNF）?α，백개소（IL）?8화전렬선소（PG）E2수평。이Western인적법검측세포Toll양수체（TLR）4、핵인자κB（NF?κB）급배양매（COX）2적표체변화。결과각과당화/혹LPS처리후Caco?2세포존활솔불수영향。여모형조상비，각과당재0.25，0.5화1.0 g/L제량하정농도의뢰성지강저Caco?2세포TNF?α（352.5±21.6，298.4±25.1，203.4±20.0 vs 436.8±38.7μg/L）화PGE2적석방（632.2±35.6，522.6±26.7，402.4±30.2 vs 822.3±23.5μg/L）（P<0.05；P<0.01），0.5화1.0 g/L각과당가강저세포COX?2표체수평（P<0.05；P<0.01）。각과당중、고농도조TLR4표체명현저우모형조（P<0.05），각과당각제량조NF?κB적표체수평현저강저（P<0.05；P<0.01）。결론각과당가대항LPS유도적장상피세포염증，대염성장병구잠재보호작용。작용궤리가능여TLR4\NF?κB신호통로억제유관。
Objective To investigate the protective effect of oligochitosan on lipopolysaccharide (LPS)induced inflammation of the intestinal epithelial cells. Methods intestinal epithelial cells Caco?2 were divided into five groups:normal control group,model group,high?,medium? and low?level oligochitosan groups. In the oligochitosan groups,the cells were pre?treated with 0.25,0.5 and 1.0 g/L oligochitosan,respectively,at 2 h before LPS stimulation. MTT method was used for cell viability assay. ELISA was used to measure the levels of inflammation related cytokines,such as tumor necrotizing factor?related factor alpha (TNF?α),interleukin?8 (IL?8) and prostaglandin E2 (PGE2),in the cell culture supernatant. Western blotting was used to detect the changes in expression of Toll?like receptor 4(TLR4), nuclear factor κB(NF?κB)and cyclooxygenase 2(COX?2). Results The Caco?2 cell viability was not affected by treatment with oligochitosan and/or LPS. Compared with the model group,0.25,0.5 and 1.0 g/L oligochitosan reduced the production of TNF?α(352.5±21.6,298.4±25.1,203.4 ±20.0 vs 436.8±38.7μg/L, P<0.05)and PGE2(632.2±35.6,522.6±26.7,402.4±30.2 vs 822.3±23.5μg/L,P<0.01)in Caco?2 cells in a dose dependent manner;0.5 and 1.0 g/L oligochitosan reduced COX 2 expression(P<0.05 and P<0.01);medium?and high?level oligochitosan was significantly associated with lowered expression of TLR 4 compared with the model group (P<0.05);the expression of NF?κB in each group of oligochitosan dosage was significantly lowered (P<0.05;P<0.01). Conclusion Oligochitosan may reverse LPS ? induced inflammation of the intestinal epithelial cells and therefore may be potentially protective against inflammatory bowel diseases. The underlysing mechanism of this action may be related to inhibition of TLR4/NF?κB signaling pathway.
