外周血单个核细胞中p38 MAPK及P-p38 MAPK在急、慢性胰腺炎炎症反应中的水平变化

Changes of p38 MAPK and P-p38 MAPK levels in peripheral blood mononuclear cells during inflammation reaction of acute and chronic pancreatitis

目的观察外周血单个核细胞(PBMC)中p38丝裂原活化蛋白激酶(p38 MAPK)及其活化形式磷酸化p38MAPK(P-p38 MAPK)在急、慢性胰腺炎炎症反应中的水平变化。方法选择2010年11月至2014年12月收治的胰腺炎患者82例,依据临床表现将患者分为慢性胰腺炎组29例、轻型急性胰腺炎组25例、重症急性胰腺炎组28例。选取同期在本院健康体检正常的37例为对照组。取静脉血,分离PBMC,采用实时聚合酶链反应(Real-time PCR)法检测各组p38 MAPK mRNA在PBMC的表达;采用Western Blot检测各组p38 MAPK、P-p38 MAPK蛋白在PBMC的表达。结果 (1)对照组、慢性组、轻型组、重症组p38 MAPK mRNA相对表达量(2-△△Ct)分别为0.33±0.08、0.35±0.04、0.34±0.07、0.42±0.05,重症组明显高于其余3组,差异有统计学意义(F=12.793,P<0.05),其中重症组均值较对照组增加了27.2%。对照组、慢性组、轻型组3组之间比较差异无统计学意义(P>0.05)。(2)p38 MAPK蛋白表达水平(相对表达量,以β-actin为内参):重症组明显高于对照组,差异有统计学意义(P<0.05),其均值较对照组增加了23.4%,对照组、慢性组、轻型组3组之间比较,差异无统计学意义(P>0.05)。P-p38 MAPK蛋白水平表达(相对表达量,以β-actin为内参):重症组P-p38 MAPK蛋白水平明显高于其他3组,差异有统计学意义(P<0.05,P<0.01),其中重症组均值较对照组、慢性组、轻型组分别增加了455.6%、51.5%、47.1%。慢性组和轻型组P-p38 MAPK蛋白水平高于对照组,差异有统计学意义(P<0.05),其均值较对照组分别增加了266.7%、277.8%。慢性组和轻型组比较,差异无统计学意义(P>0.05)。结论胰腺炎患者PBMC中p38MAPK及其磷酸化后的P-p38 MAPK,与病情的严重程度、病程的长短密切相关。对p38 MAPK和P-p38 MAPK在mRNA、蛋白水平表达的检测,有望成为重症胰腺炎临床指标和药物治疗的新靶点。

Objective To observe the levels of p38 mitogen activated protein kinase( p38 MAPK) and phosphorylated p38 MAPK( P-p38 MAPK),an activation form of p38 MAPK,in peripheral blood mononuclear cells( PBMC) during inflammation reactions of acute and chronic pancreatitis. Methods Eighty-two patients with pancreatitis admitted between November 2010 and December 2014 were selected,and the patients were divided into chronic pancreatitis group( n = 29),mild acute pancreatitis group( n = 25) and severe acute pancreatitis group( n = 28) according to clinical manifestations.Thirty-seven healthy subjects in health examination at the same time were selected as control group. Collecting venous blood and separating PBMC in all participants. The real-time PCR method was used to detect p38 MAPK mRNA expression in PBMC. The Western Blot method was used to detect the expressions of p38 MAPK and P-p38 MAPK proteins in PBMC.Results( 1) The relative expressions( 2- △△Ct) of p38 MAPK mRNA were 0. 33 ± 0. 08 in control group,0. 35 ± 0. 04 in chronic pancreatitis group,0. 34 ± 0. 07 in mild acute pancreatitis group and 0. 42 ± 0. 05 in severe acute pancreatitis group,respectively. The relative expression quantity in severe acute pancreatitis group was significantly higher than those in the other 3 groups( F = 12. 793,P < 0. 05) and increased by 27. 2% compared with control group,while there were no significant differences among control group,chronic pancreatitis group and mild acute pancreatitis group( P > 0. 05).( 2) For the p38 MAPK protein relative expression levels( β-actin for reference),the level in severe acute pancreatitis group was significantly higher than that in control group( P < 0. 05),and its mean value increased by 23. 4% compared with control group. There were no significant differences in the levels among control group,chronic pancreatitis group and mild acute pancreatitis group( P > 0. 05). For the P-p38 MAPK protein relative expression levels( β-actin for reference),the level in severe acute pancreatitis group was significantly higher than those in control group,chronic pancreatitis group and mild acute pancreatitis group( P < 0. 05,P < 0. 01),and its mean value increased by 455. 6%,51. 5% and 47. 1%,respectively compared with the other 3 groups. The levels of P-p38 MAPK protein in chronic pancreatitis group and mild acute pancreatitis group were significantly higher than that in control group,and their mean value increased by 266. 7% and 277. 8%,respectively compared with control group. There was no significant difference in the levels between chronic pancreatitis group and mild acute pancreatitis group( P > 0. 05). Conclusions The levels of p38 MAPK and P-p38 MAPK in PBMC of patients with pancreatitis were closely associated with severity of illness and duration of disease. The detection of mRNA and protein expressions of p38 MAPK mRNA and P-p38 MAPK in PBMC is expected to become a new target of drug treatment and a reference judging clinical indexes.