CAJ | 학술논문

目的探讨急性胰腺炎(AP)时胰腺腺泡细胞胀亡和巨噬细胞激活的关系.方法以两步酶消化法分离大鼠胰腺腺泡细胞,分为3组.AP组以蛙皮素制成AP细胞模型,实验组以蛙皮素+内皮素处理,对照组加等量培养液.收集腺泡细胞以吖啶橙+溴乙锭荧光染色检测细胞胀亡的情况.收集培养液部分上清液进行淀粉酶及LDH释放的检测;另取1 ml上清液加入接种分离的大鼠腹腔巨噬细胞的培养板中培养6 h,然后以ELISA法检测TNF-α蛋白水平.结果对照组细胞发生胀亡极少,胰腺腺泡细胞释放胰淀粉酶、LDH和巨噬细胞释放TNF-α也处于较低水平,分别为(1175±165)kU/L、(846±118)U/L、(36±5)μg/L;AP组可见细胞胀亡,胰淀粉酶、LDH的释放及TNF-α的水平均明显升高,分别为(7130±680)kU/L、(4262±626)U/L、(155±18)μg/L,与对照组比较差异有统计学意义(t=5.184,4.277,3.665,P<0.05);实验组各指标水平增高更明显,分别为(9240±1177)kU/L、(6937±893)U/L、(268±35)μg/L,与AP组比较差异有统计学意义(t=2.251,2.825,2.843,P<0.05).结论随着细胞胀亡的发生,胰腺腺泡细胞淀粉酶的释放增加,巨噬细胞释放TNF-α的变化趋势与细胞胀亡程度一致.巨噬细胞的炎症反应、胰腺腺泡细胞内容物的释放与胰腺腺泡细胞胀亡之间存在着联系.
목적 탐토급성이선염(AP)시이선선포세포창망화거서세포격활적관계.방법 이량보매소화법분리대서이선선포세포,분위3조.AP조이와피소제성AP세포모형,실험조이와피소+내피소처리,대조조가등량배양액.수집선포세포이아정등+추을정형광염색검측세포창망적정황.수집배양액부분상청액진행정분매급LDH석방적검측;령취1 ml상청액가입접충분리적대서복강거서세포적배양판중배양6 h,연후이ELISA법검측TNF-α단백수평.결과대조조세포발생창망겁소,이선선포세포석방이정분매、LDH화거서세포석방TNF-α야처우교저수평,분별위(1175±165)kU/L、(846±118)U/L、(36±5)μg/L;AP조가견세포창망,이정분매、LDH적석방급TNF-α적수평균명현승고,분별위(7130±680)kU/L、(4262±626)U/L、(155±18)μg/L,여대조조비교차이유통계학의의(t=5.184,4.277,3.665,P<0.05);실험조각지표수평증고경명현,분별위(9240±1177)kU/L、(6937±893)U/L、(268±35)μg/L,여AP조비교차이유통계학의의(t=2.251,2.825,2.843,P<0.05).결론 수착세포창망적발생,이선선포세포정분매적석방증가,거서세포석방TNF-α적변화추세여세포창망정도일치.거서세포적염증반응、이선선포세포내용물적석방여이선선포세포창망지간존재착련계.
Objective To study the relationship between the oncosis of pancreatic acinar cells and activa-tion of macrophage in rat model of acute pancreatitis (AP). Methods The pancreatic acinar cells were isolated by two-step enzyme digestion, and then they were divided into control group, AP group and test group. Pancreatic acinar cells were cultured with caerulein in AP group, with caerulein and endothelin in test group, and with culture medium in control group. The oncesis rate of the pancreatic acinar cells was detected after acridine orange and ethidium bromide fluorescent staining. The supernatant was collected to detect the release of amylase and lactate dehydrogenase (LDH). The macrophages were cultured with 1 ml of supematant for 6 hours, and then the protein level of tumor necrosis factor-α (TNF-α) was measured by ELISA. Results Few oncotic pancreatic acinar cells were observed in the control group, and the levels of amylase and LDH secreted by pancreatic acinar cells and TNF-α secreted by macrophage were (1175±165)kU/L, (846±118)U/L and (36±5)μg/L, respectively. Oncotic pancreatic acinar cells were observed in AP group, and the levels of amylase, LDH and TNF-α were (7130±680) kU/L, (4262±626) U/L and (155±18) μg/L, respectively, which were significantly higher than those in control group (t = 5.184, 4.277, 3.665, P < 0.05). The levels of amylase, LDH and TNF-α were even higher in test group, and they were (9240±1177) kU/L, (6937±893)U/L and (268±35)μg/L, respectively, which were significantly higher than those in AP group (t = 2.251, 2.825, 2.843, P < 0.05). Conclusions The release of amylase was changed as the oncosis of pancreatic acinar cells occurred. The secretion of TNF-α was along with the degree of oncosis of pancreatic acinar cells. The results of the study indicate that a relationship exists among the inflammatory response of macrophage, the release of contents of pancreatic acinar cells and the oncosis of the pancreatic acinar cells.