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。
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.
出处
《临床急诊杂志》
CAS
2017年第12期929-932,共4页
Journal of Clinical Emergency
