Objective: To investigate the mechanism of growth factor receptor bound protein 2(GRB2) gene silencing on lipid deposition and inflammatory infiltration in atherosclerosis(AS) rats through the regulation of PI3K/AKT/NF-κB signaling pathway. Methods: Normal saline group was set as control group, AS model rats were constructed and divided into four groups:negative control group(transfected with negative control sequence of GRB2 shRNA), GRB2 shRNA group(transfected with GRB2 shRNA sequence), pathway inhibitor group(treated with PI3K/AKT pathway inhibitor), combined group(treated with GRB2 shRNA sequence combined with PI3K/AKT pathway inhibitor). The expression of PI3KR1, AKT2 and NF-κB p65 in aorta of rats in each group was detected by quantitative real time polymerase chain reaction(qRT-PCR) and Western blotting. The morphology of aorta was observed by HE staining. Oil red O staining was used to quantify lipid deposition in the arteries of rats in each group. Enzyme-linked immunosorbent assay(ELISA) was utilized to determine the level of tumor necrosis factor α (TNF-α), C-reactive protein(CRP), lipoprorein-assoeiated phosPhohPaseA2(Lp-PLA2) in peripheral blood of rats in each group. Results: qRT-PCR and Western blotting experiments showed that PI3K/AKT signaling pathway was significantly activated in AS rats compared with saline group(P<0.05). Compared with the negative control group, the expression of PI3KR1, NF-κB p65, AKT decreased significantly after GRB2 silencing or PI3K/AKT pathway inhibition(all P<0.05). The results of HE staining indicated that AS pathological state was obvious in negative control group, the pathological condition was improved after GRB2 silencing or PI3K/AKT signaling pathway inhibition, combined group showed more obvious improvement. Oil red O staining showed that compared with negative control group, the area of lipid deposition in GRB2 shRNA group, pathway inhibitor group and combined group decreased significantly (all P<0.05). ELISA results indicated that compared with negative control group, silencing of GRB2, inhibition of PI3K/AKT pathway or combined with the above two treatments could significantly down-regulate the levels of TNF-α, CRP and Lp-PLA2 in priipheral blood in each group(all P<0.05). Conclusion: Down-regulation of GRB2 can lessen the lipid deposition and inflammatory infiltration of AS model rats through inhibiting PI3K/AKT/NF-κB signaling pathway, GRB2 gene silencing exerts protective effects in AS. |
[1] 邢晓林,刘昊凌.骨钙素对动脉粥样硬化作用的研究进展[J].现代医学,2018,46(6):732-735.
[2] WANG H T,WANG Z Z,WANG Z C,et al.Patchouli alcohol attenuates experimental atherosclerosis via inhibiting macrophage infiltration and its inflammatory responses[J].Biomed Pharmacother,2016,83(5):930-935.
[3] LIN C Y,CHEN J H,FU R H,et al.Induction of Pi form of glutathione S-transferase by carnosic acid is mediated through PI3K/Akt/NF-κB pathway and protects against neurotoxicity[J].Chem Res Toxicol,2014,27(11):1958-1966.
[4] YAN H,MA Y,LI Y,et al.Insulin inhibits inflammation and promotes atherosclerotic plaque stability via PI3K-Akt pathway activation[J].Immunol Lett,2016,170(12):7-14.
[5] 谢盈彧,许晓敏,张军平,等.基于PI3K/Akt/mTOR通路探讨补肾抗衰片介导自噬调控动脉粥样硬化的机制[J].中国中西医结合杂志,2018,38(5):586-593.
[6] JOO J H,OH H,KIM M,et al.NADPH oxidase 1 activity and ROS generation are regulated by Grb2/Cbl-mediated proteasomal degradation of NoxO1 in colon cancer cells[J].Cancer Res,2016,76(4):855-865.
[7] 王竟悟,于杨,王大新,等.血管平滑肌细胞与动脉粥样硬化关系的研究新进展[J].实用医学杂志,2017,33(21):3661-3663.
[8] 林春龙,张珍祥,谭章.低氧状态下黏着斑激酶对人肺动脉平滑肌细胞增殖的影响机制[J].中华医学杂志,2011,91(32):2274-2277.
[9] 王园园,龙民慧,邹民吉,等.大鼠动脉粥样硬化动物模型的建立和评价[J].中国实验动物学报,2008,16(6):421-423.
[10] XU Y,XU X,JIN H Y,et al.Effects of a thrombomodulin-derived peptide on monocyte adhesion and intercellular adhesion molecule-1 expression in lipopolysaccharide-induced endothelial cells[J].Mol Vis,2013,19(2):203-212.
[11] 蔡伟,张秀伟,张希龙,等.阻塞性睡眠呼吸暂停综合征、炎症因子与动脉粥样硬化间关系的研究进展[J].东南大学学报(医学版),2015,34(1):159-162.
[12] PLUMP A S,SMITH J D,HAYEK T,et al.Severe hypercholesterolemia and atherosclerosis in apolipoprotein E-deficient mice created by homologous recombination in ES cells[J].Cell,1992,71(2):343-353.
[13] 李安毅,郭晓汐,刘莹,等.支架蛋白Gab的结构与功能及其在肿瘤、炎症和心血管疾病发生中的作用[J].生理学报,2016,68(1):57-64.
[14] 闫博阳,赵津璋,陈虹.PTEN在动脉粥样硬化中的表达及对血管内皮细胞增殖凋亡的影响研究[J].中国免疫学杂志,2018,34(5):111-116.
[15] 王宇,杨欣,李晓冬,等.雌激素抑制内质网应激引起的血管内皮细胞凋亡及机制[J].中国动脉硬化杂志,2016,24(3):217-223.
[16] 付海燕,胡占升,杜红阳.白藜芦醇衍生物TMS对脂多糖诱导血管内皮细胞表达NO、ICAM-1和NFκB的影响[J].山东大学学报(医学版),2015,53(11):10-15.
[17] 胡文君,张振,戴敏.丹皮酚通过抑制PI3K/AKT-NF-κB通路对LPS诱导的与平滑肌细胞共培养的大鼠血管内皮细胞的保护作用[J].中国中药杂志,2016,41(12):2298-2302.
[18] 董晓柳,朱丽霞,徐士军.普罗布考联合瑞舒伐他汀对脑梗死合并糖尿病患者颈动脉粥样硬化斑块、血脂及炎症因子的影响[J].中国动脉硬化杂志,2016,24(2):177-181.
[19] 陈英,张文玲,黄涛,等.炎症因子TNF-α、IL-6、IL-17与类风湿关节炎并发动脉粥样硬化的关系[J].免疫学杂志,2017,33(3):268-272.
[20] 李凤,朱余友,杨孙凤,等.血清五聚素3、超敏C反应蛋白、脂蛋白相关磷脂酶A2水平以及微栓子信号与大动脉粥样硬化型急性脑梗死患者颈动脉粥样硬化斑块稳定性的相关研究[J].临床神经病学杂志,2016,29(2):95-100. |