Simvastatin protect RLE-6TN cells from LPS induced injury via HO-1 up-regulation through PI3K/AKt pathway
-
摘要: 目的:观察辛伐他丁(SIM)对脂多糖(LPS)诱导的大鼠肺泡Ⅱ型上皮细胞(RLE-6TN)损伤的保护作用及机制。方法:以离体培养RLE-6TN细胞为研究对象,观察SIM对RLE-6TN细胞血红素加氧酶-1(HO-1)表达的影响及机制。在HO-1表达最高时间点建立LPS炎症损伤模型,通过检测细胞活力和培养液中乳酸脱氢酶(LDH)含量,观察SIM对LPS诱导的RLE-6TN细胞炎症损伤的保护作用。结果:与正常对照组相比,SIM组Akt磷酸化水平显著增加,并在处理后2h达到峰值。SIM可显著上调RLE-6TN细胞HO-1表达,mRNA和蛋白分别在处理后6h和12h到达峰值。PI3K/Akt抑制剂LY294002可显著抑制SIM对RLE-6TN细胞HO-1的上调作用。LPS处理可降低RLE-6TN细胞活力和增加培养液中LDH含量。SIM可显著减轻LPS对RLE-6TN的损伤,但这种保护作用可以被HO-1抑制剂Zn-pp显著抑制。结论:SIM可通过PI3K/Akt通路上调HO-1表达并对抗LPS诱导的RLE-6TN细胞损伤。Abstract: Objective:To observe the protective effects and mechanisms of Simvastatin (SIM) on lipopolysaccharide (LPS) induced alveolar epithelial cells type Ⅱ (RLE-6TN) injury.Method:The expression of HO-1 and the phosphorylation of Akt after SIM treatment in RLE-6TN were detected by RT-QPCR and/or western blot.Using lactate dehydrogenase release assay and cell counting kit-8 assay, the injury induced by LPS was determined and compared among RLE-6TN treated with SIM with or without HO-1 inhibitor.Result:The phosphorylation of Akt after SIM treatment was significantly increased.RT-QPCR results showed that the mRNA levels of HO-1 significantly increased and reached highest at 6 h and western blot further showed that HO-1 significantly increased and reached highest at 12 hfollowing SIM treatment and reversed by PI3K/Akt inhibitor LY294002.SIM significantly increased the cell viability and decreased the medium lactate dehydrogenase content in cultures treated with LPS.Pretreatment with zinc protoporphyrin IX, a specific inhibitor of HO-1, significantly blocked the protective effects of SIM.Conclusion:These results suggested that SIM could protect RLE-6TN against LPS injury mostly by up-regulating of HO-1 expression through PI3K/Akt pathway.
-
Key words:
- Simvastatin /
- RLE-6TN /
- heme oxygenase-1 /
- PI3K/Akt
-
[1] Standiford T J, Ward P A.Therapeutic targeting of acute lung injury and acute respiratory distress syndrome[J].Translational Res:J Lab Clin Med, 2016, 167:183-191.
[2] 李成恩, 郝建, 李树雯, 等.爆炸致家兔急性肺损伤及乌司他丁的保护作用[J].中国急救医学, 2016, 36 (9):842-845.
[3] Hsu H H, Ko W J, Hsu J Y, et al.Simvastatin ameliorates established pulmonary hypertension through a heme oxygenase-1dependent pathway in rats[J].Respiratory Res, 2009, 10:32-32.
[4] Jang H J, Hong E M, Kim M, et al.Simvastatin induces heme oxygenase-1 via NF-E2-related factor 2 (Nrf2) activation through ERK and PI3K/Akt pathway in colon cancer[J].Oncotarget, 2016.
[5] Zhao W, Song H, Huo W.Long-term administration of simvastatin reduces ventilator-induced lung injury and upregulates heme oxygenase 1 expression in a rat model[J].J Surg Res, 2015, 199:601-607.
[6] Lee T S, Chang C C, Zhu Y, et al.Simvastatin induces heme oxygenase-1:a novel mechanism of vessel protection[J].Circulation, 2004, 110:1296-1302.
[7] Zhang F, Sun D, Chen J, et al.Simvastatin attenuates angiotensin IIinduced inflammation and oxidative stress in human mesangial cells[J].Mol Med Rep, 2015, 11:1246-1251.
[8] Zhao Y, Feng Q, Huang Z, et al.Simvastatin inhibits inflammation in ischemia-reperfusion injury[J].Inflammation, 2014, 37:1865-1875.
[9] Barnett M, Hall S, Dixit M, et al.Simvastatin attenuates oleic acid-induced oxidative stress through CREBdependent induction of heme oxygenase-1 in renal proximal tubule cells[J].Pediatric Research, 2016, 79:243-250.
[10] Richards J A, Wigmore S J, Devey L R.Heme oxygenase system in hepatic ischemia-reperfusion injury[J].World J Gastroenterol:WJG, 2010, 16:6068-6078.
[11] Boczkowski J, Poderoso J J, Motterlini R.CO-metal interaction:Vital signaling from a lethal gas[J].Trends in biochemical sciences, 2006, 31:614-621.
[12] Hayashi S, Takamiya R, Yamaguchi T, et al.Induction of heme oxygenase-1suppresses venular leukocyte adhesion elicited by oxidative stress:role of bilirubin generated by the enzyme[J].Circ Res, 1999, 85:663-671.
[13] Kim K J, Kim K S, Kim N R, et al.Effects of simvastatin on the expression of heme oxygenase-1in human RPE cells[J].Investigative Ophthalmol Visual Sci, 2012, 53:6456-6464.
计量
- 文章访问数: 66
- PDF下载数: 47
- 施引文献: 0