摘要
目的:探讨PTD-NBD多肽对体外大鼠胰腺腺泡细胞核因子-κB(nuclear factorκB,NF-κB)p65的表达影响.方法:分离培养大鼠胰腺腺泡细胞,分为正常对照组、急性胰腺炎(acute pancreatitis,AP)组和PTD-NBD多肽组,用脂多糖(10mg/L)诱导体外细胞AP模型,分别于建模6和12h观察细胞形态学变化,培养液中淀粉酶(amylase,AMY)、超氧化物歧化酶(superoxide dismutase,SOD)和白介素(interleukin,IL)-1β含量,RT-PCR和免疫印迹法检测细胞中NF-κB p65RNA及其蛋白表达.结果:与对照组相比,AP组各时间点腺泡细胞发生肿胀和死亡增多(细胞形态学评分:6h:8.90±0.34vs1.10±0.13;12h:9.40±0.26vs1.20±0.15,P<0.05),培养液中AMY升高(6h:2135.8±347.2vs873.5±91.6;12h:3299.6±217.7vs917.7±101.9,P<0.05),IL-1β含量升高(6h:84.9±15.7vs39.3±7.9;12h:95.6±17.1vs38.9±5.2,P<0.05),SOD含量降低(6h:116.3±30.3vs176.2±21.6;12h:101.5±25.6vs173.6±27.9,P<0.05),细胞中NF-κB p65RNA及其蛋白表达量增多(P<0.05);与AP组比较,经PAT-NBD多肽预处理后腺泡细胞出现肿胀和死亡减少(细胞形态学评分:6h:6.80±0.23vs8.90±0.34;12h:7.50±0.19vs9.40±0.26,P<0.05),培养液中SOD上升(6h:137.6±27.4vs116.3±30.3;12h:144.3±23.6vs101.5±25.6,P<0.05),AMY下降(6h:1951.5±211.7vs2135.8±347.2;12h:1761.3±231.5vs3299.6±217.7,P<0.05),IL-1β含量明显下降(6h:66.8±11.6vs84.9±15.7;12h:54.8±21.2vs95.6±17.1,P<0.05),NF-κB p65蛋白表达降低(P<0.05).结论:PTD-NBD多肽透过大鼠胰腺腺泡细胞膜抑制脂多糖刺激的NF-κB p65活化及其活性,下调IL-1β表达,上调SOD含量,从而减轻胰腺腺泡细胞的炎症损伤.
AIM：To examine the effect of PTD-NBD polypeptide on the expression of nuclear factor B（NF-κB） p65 in inflammatory pancreatic acinar cell injury in rats.METHODS：Rat pancreatic acinar cells were isolated,cultured,and divided into a normal control group,an acute pancreatitis（AP） group and a PTD-NBD polypeptides group.An in vitro model of AP was induced by treating rat pancreatic acinar cells with lipopolysaccharide（10 mg/L）.Cell morphological changes were observed,and the contents of amylase,superoxide dismutase（SOD） and IL-1 in culture medium were tested.Expression of NF-κB p65 mRNA and protein in cells was detected by RT-PCR and Western blot 6 and 12 h after modeling,respectively.RESULTS：Compared to the control group,pancreatic acinar cell swelling and death were increased（6 h：8.9 ± 0.34 vs 1.1 ± 0.13;12 h：9.4 ± 0.26 vs 1.2 ± 0.15,both P 0.05）,the contents of amylase（6 h：2135.8 ± 347.2 vs 873.5 ± 91.6;12 h：3299.6 ± 217.7 vs 917.7 ± 101.9,both P 0.05） and IL-1（6 h：84.9 ± 15.7 vs 39.3 ± 7.9;12 h：95.6 ± 17.1 vs 38.9 ± 5.2,both P 0.05） were increased and the contents of SOD were decreased in culture medium（6 h：116.3 ± 30.3 vs 176.2 ± 21.6;12 h：101.5 ± 25.6 vs 173.6 ± 27.9,P 0.05）,and the expression of NF-κB p65 in pancreatic acinar cells was increased（P 0.05） in the AP group at 6 and 12 h after modeling.Compared to the AP group,pancreatic acinar cell swelling and death were lessened（6 h：6.8 ± 0.23 vs 8.9 ± 0.34;12 h：7.5 ± 0.19 vs 9.4 ± 0.26,both P 0.05）,the contents of SOD were raised（6 h：137.6 ± 27.4 vs 116.3 ± 30.3;12 h：144.3 ± 23.6 vs 101.5 ± 25.6,both P 0.05） and the contents of amylase（6 h：1951.5 ± 211.7 vs 2135.8 ± 347.2;12 h：1761.3 ± 231.5 vs 3299.6 ± 217.7,both P 0.05） and IL-1（6 h：66.8 ± 11.6 vs 84.9 ± 15.7;12 h：54.8 ± 21.2 vs 95.6 ± 17.1,both P 0.05） were decreased in culture medium,and the expression of NF-κB p65 mRNA and protein was down-regulated in the PAT-NBD polypeptide group（
出处
《世界华人消化杂志》
CAS
北大核心
2013年第22期2136-2142,共7页
World Chinese Journal of Digestology
基金
广东省科技计划基金资助项目
No.2007A060305007
No.2007B031405004
No.2012B060300029
广东省医学科研基金资助项目
No.B2012187
广东省中山市科技计划基金资助项目
No.20102A124~~
