| 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（10 μmol/L、50 μmol/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10 μmol/L或50 μmol/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。 |
| 关键词: 骨桥蛋白 胰腺星形细胞 PI3K/Akt信号通路 α-平滑肌肌动蛋白 |
| DOI：10.16781/j.0258-879x.2019.03.0316 |
| 投稿时间：2018-09-16修订日期：2019-01-09 |
| 基金项目: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）. |
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| Osteopontin activates pancreatic stellate cells through PI3K/Akt signaling pathway |
| 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
*Corresponding author) |
| Abstract: |
| 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. |
| Key words: osteopontin pancreatic stellate cells PI3K/Akt signaling pathway α-smooth muscle actin |
