CaMKⅡ在小鼠重症急性胰腺炎胰腺损伤中的作用

Role of CaMK Ⅱ in pancreatic injury in mice with severe acute pancreatitis

目的观察钙调素依赖蛋白激酶(CaMKⅡ)在重症急性胰腺炎(SAP)小鼠胰腺组织中的表达情况,并探讨其抑制剂KN93对SAP胰腺损伤的保护作用和机制。方法36只健康雄性C57小鼠随机分为假手术组、SAP组、KN93组、SAP组+KN93组,9只/组。于造模后24 h收集血清和胰腺组织;采用HE染色观察胰腺组织病理改变;Elisa检测血清脂肪酶、淀粉酶活性以及炎性因子变化情况;免疫印迹法检测CaMKⅡ、p-CaMKⅡ、p-NF-κB、MAPK、p-MAPK在小鼠胰腺组织中的表达变化。结果与假手术组比较,SAP组p-CaMKⅡ、p-NF-κB、p-MAPK表达水平升高(P<0.05)。KN93干预后,SAP小鼠胰腺组织病理损伤减轻,血清脂肪酶、淀粉酶以及炎性因子(TNF-α、IL-6)水平降低(P<0.05),同时NF-κB、ERK和MAPK蛋白磷酸化也降低(P<0.05)。结论CaMKⅡ蛋白在SAP疾病胰腺组织活性升高,CaMKⅡ抑制剂KN93能有效减轻SAP小鼠胰腺损伤和炎症程度,其机制可能与ERK/MAPK信号通路有关。

Objective To investigate the expression of Ca^(2+)/calmodulin-dependent protein kinase II(CaMKⅡ) in pancreatic tissues of mice with severe acute pancreatitis(SAP) and explore the protective effect of KN93, a CaMKⅡinhibitor, against pancreatic injury in SAP and the possible mechanism. Methods Thirty-six healthy male C57 mice were randomly divided into sham operation group, SAP group, KN93 group and SAP + KN93 group(n=9). Serum and pancreatic tissue samples were collected 24 h after modeling. The pathological changes in the pancreatic tissues were observed using HE staining. Serum lipase and amylase activities and the levels of inflammatory factors were detected using ELISA. Western blotting was used to detect the expressions of CaMKⅡ, p-CaMKⅡ, p-NF-κB, MAPK and p-MAPK in mouse pancreas. Results Compared with those in sham operation group, the expressions of p-CaMKⅡ, p-NF-κB and p-MAPK were significantly increased in SAP group(P<0.05). KN93 treatment obviously alleviated pathological injuries of the pancreas in SAP mice, and significantly lowered serum levels of lipase, amylase and inflammatory factors(TNF-α and IL-6) and phosphorylation levels of NF-κB, ERK and MAPK proteins(P<0.05). Conclusion The activity of CaMKⅡis significantly increased in the pancreatic tissue of SAP mice. KN93 can alleviate pancreatic injury and inflammation in SAP mice possibly through the ERK/MAPK signaling pathway.