中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2010年
5期
522-527
,共6页
急性胰腺炎%脂多糖%胰腺腺泡AR42J细胞%核因子-κB%肿瘤坏死因子-α
急性胰腺炎%脂多糖%胰腺腺泡AR42J細胞%覈因子-κB%腫瘤壞死因子-α
급성이선염%지다당%이선선포AR42J세포%핵인자-κB%종류배사인자-α
Acute pancreatitis%Lipopolysaccharide (LPS)%AR42J cell lines%Nuclear factor-κB%Tumor necrosisi factor-α
目的 探讨脂多糖(lipopolysaccharide,LPS)对胰腺腺泡AR42J细胞细胞因子表达的影响及其可能机制,进一步阐明急性胰腺炎的发病机制.方法 用不同质量浓度的LPS(0.001,0.01,0.1,1,10,100 mg/L)刺激AR42J细胞18 h,同时用10mg/L的LPS刺激AR42J细胞不同时间(2,6,12,18,24h),RT-PCR检测核因子-κB(NF-κB)P65和肿瘤坏死因子-α(TNF-α)mRNA表达的变化,放射免疫法(RIA)检测培养液上清TNF-α蛋白浓度的改变.对NF-κB(P65)mRNA的表达与TNF-α mRNA的表达进行相关和回归分析.结果 RT-PCR结果表明0.001 mg/L的LPS处理AR42J细胞时即可出现TNF-α及NF-κB(P65)mRNA表达的明显上调,且呈量效关系;用10 mg/L的IPS处理后,在2 h后即可出现TNF-α及NF-κB(P65)mRNA表达的明显上调,并且呈时效关系.RIA结果表明用0.01 mg/L的LPS处理AR42J细胞后即可出现TNF-α蛋白表达的明显上调,且呈量效关系;用10mg/L的LPS处理后,在6 h后即可出现TNF-α蛋白表达的明显上调,并且呈时效关系.TNF-α mRNA的表达与P65 mRNA的表达呈正相关.结论 LPS可以以时间剂量依赖方式地刺激AR42J细胞NF-κB(P65)和TNF-α的表达,NF-κB(P65)mRNA的表达与TNF-αmRNA的表达呈正相关.针对NF-κB靶点,抑制其活性,可为包括AP的治疗提供一条新的途径.
目的 探討脂多糖(lipopolysaccharide,LPS)對胰腺腺泡AR42J細胞細胞因子錶達的影響及其可能機製,進一步闡明急性胰腺炎的髮病機製.方法 用不同質量濃度的LPS(0.001,0.01,0.1,1,10,100 mg/L)刺激AR42J細胞18 h,同時用10mg/L的LPS刺激AR42J細胞不同時間(2,6,12,18,24h),RT-PCR檢測覈因子-κB(NF-κB)P65和腫瘤壞死因子-α(TNF-α)mRNA錶達的變化,放射免疫法(RIA)檢測培養液上清TNF-α蛋白濃度的改變.對NF-κB(P65)mRNA的錶達與TNF-α mRNA的錶達進行相關和迴歸分析.結果 RT-PCR結果錶明0.001 mg/L的LPS處理AR42J細胞時即可齣現TNF-α及NF-κB(P65)mRNA錶達的明顯上調,且呈量效關繫;用10 mg/L的IPS處理後,在2 h後即可齣現TNF-α及NF-κB(P65)mRNA錶達的明顯上調,併且呈時效關繫.RIA結果錶明用0.01 mg/L的LPS處理AR42J細胞後即可齣現TNF-α蛋白錶達的明顯上調,且呈量效關繫;用10mg/L的LPS處理後,在6 h後即可齣現TNF-α蛋白錶達的明顯上調,併且呈時效關繫.TNF-α mRNA的錶達與P65 mRNA的錶達呈正相關.結論 LPS可以以時間劑量依賴方式地刺激AR42J細胞NF-κB(P65)和TNF-α的錶達,NF-κB(P65)mRNA的錶達與TNF-αmRNA的錶達呈正相關.針對NF-κB靶點,抑製其活性,可為包括AP的治療提供一條新的途徑.
목적 탐토지다당(lipopolysaccharide,LPS)대이선선포AR42J세포세포인자표체적영향급기가능궤제,진일보천명급성이선염적발병궤제.방법 용불동질량농도적LPS(0.001,0.01,0.1,1,10,100 mg/L)자격AR42J세포18 h,동시용10mg/L적LPS자격AR42J세포불동시간(2,6,12,18,24h),RT-PCR검측핵인자-κB(NF-κB)P65화종류배사인자-α(TNF-α)mRNA표체적변화,방사면역법(RIA)검측배양액상청TNF-α단백농도적개변.대NF-κB(P65)mRNA적표체여TNF-α mRNA적표체진행상관화회귀분석.결과 RT-PCR결과표명0.001 mg/L적LPS처리AR42J세포시즉가출현TNF-α급NF-κB(P65)mRNA표체적명현상조,차정량효관계;용10 mg/L적IPS처리후,재2 h후즉가출현TNF-α급NF-κB(P65)mRNA표체적명현상조,병차정시효관계.RIA결과표명용0.01 mg/L적LPS처리AR42J세포후즉가출현TNF-α단백표체적명현상조,차정량효관계;용10mg/L적LPS처리후,재6 h후즉가출현TNF-α단백표체적명현상조,병차정시효관계.TNF-α mRNA적표체여P65 mRNA적표체정정상관.결론 LPS가이이시간제량의뢰방식지자격AR42J세포NF-κB(P65)화TNF-α적표체,NF-κB(P65)mRNA적표체여TNF-αmRNA적표체정정상관.침대NF-κB파점,억제기활성,가위포괄AP적치료제공일조신적도경.
Objective To investigate lipopolysaccharide (LPS) stimulates the expression of nuclear factor-KB (NF-κB) mRNA and tumor necrosis factor-a( TNF-α) mRNA in rat's pancreatic acinus AR42J cell line, and further address the pathogenesis of acute pancreatitis. Method The AR42J cell line was stimulated with different concentrations of LPS (0.001, 0.01, 0.1, 1.0, 10 and 100 mg/L) for 18 hours, or stimulated with 10 mg/L of LPS for different lengths of time (2, 6, 12,18 and 24 hrs) .Then, the expressions of NF-κB-P65 mRNA and TNF-α mRNA were determined by using RT-PCR, the levels of TNF-α protein in the culture supernatant were measured with radio-immuno assay (RIA), and the correlation between the expressions of TNF-α mRNA and NF-κB mRNA was analyzed. Results Both the expressions of NF-κB mRNA and TNF-α mRNA were up-regulated when AR42J cell line was stimulated with 10 mg/L of LPS for 2 hours or with 0.001 mg/L of LPS for 18 hours in both dose-dependent and time-dependent manners. Similarly, the levels of TNF-α protein were up-regulated when AR42J cell line was stimulated with 0.01 mg/L of LPS for 18 hours or with 10 mg/L of LPS for 6 hours in both dose-dependent and time-dependent manners. Statistical analysis revealed the positive correlation between the expressions of TNF-α mRNA and NF-κB-P65 mRNA(r = 0.962, P < 0.01). Conclusions LPS stimulates the expressions of TNF-α mRNA and NF-κB mRNA in both dose-dependent and time-dependent manners, and their expressions are closely correlated, suggesting the inhibition of their expressions as a potential therapeutic target for acute panceatitis.
