天麻素通过调节miRNA-221/PI3K/Akt通路对LPS诱导的肺泡上皮细胞损伤的影响  

Effect of gastrodin on alveolar epithelial cells injury induced by lipopolysaccharide through miRNA-221/PI3K/Akt pathway.

作　　者：杨亚勤[1] 孙冰[1] 牛丽丹[1] 张江波 张月华 谢青 杨飞云[1] YANG Ya-qin;SUN Bing;NIU Li-dan;ZHANG Jiang-bo;ZHANG Yue-hua;XIE Qing;YANG Fei-yun(Department of Emergency Medicine,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan,CHINA)

机构地区：[1]新乡医学院第一附属医院急救医学科,河南卫辉453100

出　　处：《海南医学》2023年第24期3497-3501,共5页Hainan Medical Journal

基　　金：国家自然科学基金青年基金(编号:81502313)。

摘　　要：目的探索天麻素(gastrodin,GAS)及磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)通路在脂多糖(LPS)诱导的肺泡上皮细胞损伤中的作用及可能机制。方法将人肺泡上皮细胞A549分为正常对照组、LPS组、GAS组、miR-221 siRNA组、control siRNA组,采用RT-qPCR实验检测miRNA-221 mRNA的表达水平;ELISA实验检测细胞培养液中炎性因子的含量;MTT实验检测细胞活力;Western blot实验检测细胞中磷脂酰肌醇3-激酶(PI3K)、磷酸化蛋白激酶B(p-Akt)、蛋白激酶B(Akt)、Bcl-2相关X蛋白(Bax)、B细胞淋巴瘤基因-2(Bcl-2)及半胱氨酸/天冬氨酸蛋白酶9(Caspase9)蛋白的表达情况。结果与LPS组比较,GAS组中GAS可明显降低LPS诱导的A549细胞中miRNA-221 mRNA的表达,升高p-Akt蛋白的表达,差异均具有统计学意义(P<0.05);与LPS组比较,miRNA-221 siRNA组抑制miRNA-221的表达后,可促进LPS诱导的A549细胞中PI3K、p-Akt蛋白的表达,差异均具有统计学意义(P<0.05);与LPS组比较,miRNA-221 siRNA组与GAS组均可明显降低LPS诱导的A549细胞培养液中肿瘤坏死因子α(TNF-α),白细胞介素6(IL-6),白细胞介素1β(IL-1β)的含量,降低促凋亡蛋白Bax及Caspase-9的表达,促进抑凋亡蛋白Bcl-2的表达,差异均具有统计学意义(P<0.05);而miRNA-221 siRNA组与GAS组对细胞的作用比较差异无统计学意义(P>0.05)。结论GAS通过抑制miR-221的表达,激活PI3K/Akt信号通路,进而减少炎性因子的释放,减少细胞凋亡,提高细胞活力,发挥细胞保护作用。Objective To explore the role and possible mechanism of gastrodin(GAS)and phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)pathway in alveolar epithelial cells injury induced by lipopolysaccharide(LPS).Methods Human alveolar epithelial cells A549 were randomly divided into normal control group,LPS group,GAS group,miR-221 siRNA group,control siRNA group.The expression of miRNA-221 mRNA was detected by RT-qPCR.The concentration of inflammatory factors were detected by ELISA assay.The cell viability was detected by MTT assay.The expression of phosphatidylinositol 3-kinase(PI3K),phosphorylated protein kinase B(p-Akt),protein kinase B(Akt),Bcl-2 associated X protein(Bax),B-cell lymphoma-2(Bcl-2)and cysteine-requiring aspartate protease 9(Caspase-9)proteins were detected by Western blot.Results Compared with LPS group,GAS(GAS group)significantly decreased the expression of miRNA-221 mRNA and increased the expression of p-Akt protein in A549 cells induced by LPS(P<0.05).Compared with LPS group,inhibition of miRNA-221 expression(miRNA-221 siRNA group)could increase the expressions of PI3K and p-Akt proteins in A549 cells induced by LPS(P<0.05).Compared with LPS group,miRNA-221 siRNA(miRNA-221 siRNA group)and GAS(GAS group)could significantly reduce the concentration of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β)in A549 cell culture medium,decrease the expression of pro-apoptotic proteins Bax and Caspase-9,promote the expression of anti-apoptotic protein Bcl-2 in A549 cells induced by LPS(P<0.05).There was no statistically significant difference in the effect on the cells between miRNA-221 siRNA group and GAS group(P>0.05).Conclusion GAS plays a cytoprotective role by inhibiting the expression of miR-221 and activating PI3K/Akt signaling pathway,thereby reducing the release of inflammatory factors,reducing cell apoptosis,and increasing cell viability.

关 键 词：天麻素 miRNA-221 信号通路 细胞损伤 炎性因子 作用机制 

分 类 号：R563[医药卫生—呼吸系统]