沉默LncRNA MALAT1对补脾益气法调节DE大鼠海马中I-κB、IL-6和PI3K表达的影响

Effect of Silencing LncRNA MALAT1 on Regulating Expressions of I-κB,IL-6 and PI3K in Hippocampus of DE Rats by Tonifying Spleen and Qi Effects Method

目的 探讨将沉默肺腺癌转移相关转录本1 (metastasis associated lung adenocarcinoma transcript 1,MALAT1)做为影响因素,中药方剂归脾汤对糖尿病脑病(diabetic encephalopathy, DE)大鼠海马相关信号转导通路中核因子-κB(NF-κB/I-κB)、炎性因子白细胞介素-6 (IL-6)以及磷脂酰肌醇-3-激酶(PI3K)等相对表达的影响。方法 以高脂饲料结合STZ复制并筛选出DE大鼠模型;分为对照组、模型组,沉默LncRNA MALAT1同时应用归脾汤组(M归脾汤组),归脾汤组,西药组,并应用氧化酶法测其血糖值,采用ELISA法测其胰岛素水平后计算对应的胰岛素敏感指数,选择RT-PCR法检测其海马中I-κB和IL-6 mRNA的相对表达;选用WB法检测其海马中I-κB、IL-6、PI3K蛋白的相对表达。结果 在检测各海马样本中IL-6、I-κB和PI3K相对表达水平时发现,药物治疗4周后,模型组与对照组比较:IL-6表达水平明显上升,I-κB和PI3K表达水平显著下降,差异有统计学意义(P<0.05),提示DE大鼠海马组织中的NF-κB/PI3K信号通路被激活;而在归脾汤组、M归脾汤组与模型组比较后得出:IL-6表达水平明显下降,I-κB和PI3K表达水平显著上升,差异有统计学意义(P<0.05),表明归脾汤能够调节NF-κB/PI3K信号通路,缓解DE大鼠海马组织中的炎性反应,利于改善病情。而归脾汤协同沉默MALAT1可经由进一步下调IL-6,上调IκB-和PI3K水平,更为显著抑制DE大鼠海马组织中NF-κB/PI3K通路的激活,减轻机体炎性反应。结论 中药方剂归脾汤可以调节DE大鼠模型的胰岛素抵抗状况,此效果可能是经由调节I-κB、IL-6和PI3K的表达来实现的,且沉默MALAT1后能加强归脾汤对上述指标的作用,从而为防治DE提供实验基础。

Objective The purpose of this study was to investigate the effect of Guipi Decoction(归脾汤)on the relative expressions of inhibitor of NF-κB(I-κB),interleukin-6(IL-6)and phosphatidylinositol-3-kinase(PI3 K)in hippocampal related signal transduction pathway in diabetic encephalopathy(DE)rats,taking the silencing of metastasis associated lung adenocarcinoma transcript 1(MALAT1)as an influencing factor.Methods The DE rat model was reproduced and screened by high-fat diet combined with streptozotocin(STZ).They were divided into control group,model group,silence LncRNA MALAT1 and Guipi Decoction group,Guipi Decoction group and Western medicine group.The blood glucose was measured by oxidase method.After measuring the insulin level by ELISA,the corresponding insulin sensitivity index was calculated.The relative expressions of I-κB and IL-6 mRNA in hippocampus were detected by RT-PCR.Western blot method was used to detect the relative expressions of I-κB,IL-6 and PI3 K protein in hippocampus.Results In this study,the relative expression levels of IL-6,I-κB and PI3 K in each hippocampal sample were detected.After 4 weeks of drug treatment,the expression level of IL-6 in the model group was increased significantly and the expression levels of I-κB and PI3 K were decreased significantly compared with those of the control group.The difference was statistically significant(P<0.05),suggesting that the NF-κB/PI3 K signal pathway in the hippocampus of DE rats was activated.Compared with those of the model group,the expression levels of IL-6 were decreased significantly,and the expression levels of I-κB and PI3 K were increased significantly(P<0.05),indicating that Guipi Decoction can regulate the NF-κB/PI3 K signal pathway,alleviate the inflammatory reaction in the hippocampus of DE rats and relieve the disease.The synergistic silencing of MALAT1 by Guipi Decoction can further down-regulate the levels of I-κB,IL-6 and PI3 K,significantly inhibit the activation of NF-κB/PI3 K pathway in the hippocampus of DE rats and reduce the inflammatory response.Conclusion Guipi Decoction can regulate the insulin resistance of DE rat model.This effect may be achieved by regulating the expressions of I-κB,IL-6 and PI3 K,and silencing MALAT1 can strengthen the effect of Guipi Decoction on the above indexes,so as to provide an experimental basis for the prevention and treatment of DE.