摘要
目的 观察糖尿病/星型胶质细胞富集磷蛋白酶-15（PED/PEA-15）的表达对蛙皮素诱导的体外急性胰腺炎（AP）模型腺泡细胞凋亡的影响.方法 建立体外AP模型：用10 nmol/L的蛙皮素处理胰腺腺泡细胞株（AR42J细胞）建立AP模型,4h后检测培养液淀粉酶水平,实时荧光定量反转录聚合酶链反应（RT-PCR）检测PED/PEA-15的mRNA表达,吖啶橙/溴乙锭（AO/EB）法检测AR42J细胞的凋亡.构建PED/PEA-15的真核表达载体pcDNA3和空载体,利用脂质体转染AR42J细胞,72 h后加入10 nmol/L的蛙皮素处理,RT-PCR检测PED/PEA-15的mRNA表达水平,AO/EB法检测AR42J细胞的凋亡.结果 蛙皮素处理的AR42J细胞培养液淀粉酶水平较未处理细胞明显升高[（748.75±42.90） U/L较（249.75±27.16） U/L,P＜0.05],PED/PEA-15的mRNA表达降低（5.24±0.66）倍（P＜0.05）,细胞凋亡率[（78.75±0.03）％]较未处理组[（17.50±0.04）％]升高（P＜0.05）.PED/PEA-15-pcDNA组AR42J细胞较空载体组PED/PEA-15的mRNA表达升高（4.27±0.78）倍（P＜0.05）,淀粉酶水平[（528.71±34.92） U/L]较空载体组[（856.29±52.39） U/L]明显下降（P＜0.05）,细胞凋亡率[（22.19±1.21）％]较空载体组[（68.92±1.83）％]降低（P＜0.05）,转染空载体组各项指标与未转染组差异无统计学意义（P＞0.05）.结论 PED/PEA-15蛋白在胰腺腺泡细胞的表达升高,能够抑制蛙皮素诱导的AR42J胰腺腺泡细胞凋亡.
Objective To investigate the effect of diabetes mellitus/astrocyte enriched phosphorus protease 15 （PED/PEA-15） expression on the apoptosis of acinar cells in bombesin-induced acute pancreatitis.Methods A model of acute pancreatitis with bombesin of 10 nmol/L was established.Amylase was determined 4 h later.The real-time reverse transcription polymerase chain reaction （RT-PCR） was used to detect the expression of PED/PEA-15 mRNA.The pcDNA or empty vehicle was created and transfected into AR42J cells.After the AR42J cells were treated with 10 nmol/L bombesin for 72 h, the expression of PED/PEA-15 mRNA was detected, and the apoptosis of pancreatic acinar cells was assayed by acridine orange / ethidium bromide （AO/EB） method.Results The expression of PED/PEA-15 in acute pancreatitis was decreased （5.24 ± 0.66） times compared to the control groups （P 〈 0.05）.The expression of PED/PEA-15 induced a （4.27 ± 0.78）-fold increase after transfection of PED/PEA-15-pcDNA compared to empty vehicle group （P 〈 0.05）, in the same time the apoptosis rate of acinar cells was decreased [（22.19±1.21）% vs.（68.92±1.83）%, P〈0.05].Conclusion The overexpression of PED/PEA-15 can inhibit the apoptosis of pancreatic acinar cells in acute pancreatitis.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2015年第11期2655-2657,共3页
Chinese Journal of Experimental Surgery
