摘要
目的探讨骨桥蛋白(OPN)对胰腺星形细胞(PSC)的影响及其机制。方法构建OPN慢病毒过表达载体(OPN-O/E)并转染PSC,设置空载体转染细胞作为对照组。分别采用CCK-8实验和Transwell小室检测OPN-O/E转染及OPN-O/E转染联合Akt抑制剂LY294002(10μmol/L、50μmol/L)处理后PSC增殖活性和趋化活性,采用蛋白质印迹法检测α-平滑肌肌动蛋白(α-SMA)及PI3K/Akt信号通路相关蛋白的表达。结果 OPN-O/E转染上调OPN表达后,PSC增殖活性增高、趋化活性增高,细胞中磷酸化PI3K(p-PI3K)、磷酸化Akt(p-Akt)和α-SMA表达水平均增高,与对照组比较差异均有统计学意义(P<0.05或P<0.01);细胞中PI3K和Akt表达水平与对照组相比差异均无统计学意义(P均>0.05)。使用10μmol/L或50μmol/L Akt抑制剂LY294002干预后,细胞中α-SMA及p-Akt的表达被抑制,与OPN-O/E组相比差异均有统计学意义(P均<0.01),Akt的表达无明显变化。结论 OPN通过PI3K/Akt信号通路介导PSC活化,活化后PSC的增殖及趋化活性也增强。
Objective To explore the effect of osteopontin (OPN) on pancreatic stellate cell (PSC) and its mechanisms. Methods We transfected PSC with OPN lentiviral overexpression vector (OPN-O/E) and constructed empty vector control cells (control group). After PSCs were treated with OPN-O/E or OPN-O/E in combination with Akt inhibitor LY294002 (10 μmol/L and 50 μmol/L), the proliferation ability and chemotactic activity were detected by CCK-8 and Transwell assays, respectively. The expression levels of α-smooth muscle actin (α-SMA) and related proteins of PI3K/Akt signal pathway were determined by Western blotting. Results Compared with the control group, proliferation ability and chemotactic activity of PCS were significantly increased in the OPN-O/E group, and the expression levels of phosphorylated- PI3K (p-PI3K), phosphorylated-Akt (p-Akt) and α-SMA were also significantly increased (P<0.05 or P<0.01). There were no significant differences in the expression levels of PI3K or Akt between the OPN-O/E and control groups (both P>0.05). Compared with the OPN-O/E group, the expression levels of α-SMA and p-Akt were significantly inhibited in the PCS treated with 10 μmol/L or 50 μmol/L LY294002 (all P<0.01);however, there was no significant difference in the Akt expression. Conclusion OPN can activate PSC through the PI3K/Akt signaling pathway, and the proliferation ability and chemotactic activity of activated PSC are also increased.
作者
安薇
施新岗
朱建伟
李桂香
孙畅
AN Wei;SHI Xin-gang;ZHU Jian-wei;LI Gui-xiang;SUN Chang(Department of Gastroenterology,Changhai Hospital,Naval Medical University (Second Military Medical University),Shanghai 200433,China)
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2019年第3期316-320,共5页
Academic Journal of Second Military Medical University
基金
国家自然科学基金(81400669
81470885)
上海市卫生和计划生育委员会科研课题(20144Y0255)
第二军医大学青年启动基金(2013QN13)~~
