2023 年8 期 第31 卷
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氧化应激环境下 LncRNA MALAT1 对内皮细胞TLR4/MyD88/NF-κB 信号通路的影响
Effect of LncRNA MALAT1 on TLR4/MyD88/NF-κB Signaling Pathway in Endothelial Cells under Oxidative Stress
作者:纪海涛,赵颖馨,于锡巧,柴强,刘振东,张丛丛
- 单位:
- 1.250000山东省济南市,山东第一医科大学临床与基础医学院 2.250000山东省济南市章丘区口腔医院牙体牙髓科
- Units:
- 1.School of Clinical and Basic Medicine, Shandong First Medical University, Jinan 250000, China2.Department of Endodontics, Jinan Zhangqiu Stomatological Hospital, Jinan 250000, ChinaCorresponding author: ZHAO Yingxin, E-mail: xiaoying781013@126.com
- 关键词:
- 氧化性应激;长链非编码RNA肺腺癌转移相关转录物1;Toll样受体4;髓样分化因子88;NF-κB
- Keywords:
- Oxidative stress; Long chain non-coding RNA metastasis-associated lung adenocarcinoma tran 1; Toll-like receptor 4; Myeloid differentiation factor 88; NF-kappa B
- CLC:
- R 349.1
- DOI:
- 10.12114/j.issn.1008-5971.2023.00.214
- Funds:
- 山东省自然科学基金资助项目(ZR2020MH043)
摘要:
目的 探究氧化应激环境下长链非编码RNA肺腺癌转移相关转录物1(LncRNA MALAT1)对内皮细胞Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核因子κB(NF-κB)信号通路的影响。方法 本实验时间为2022年10—12月。将人脐静脉内皮细胞(HUVEC)分为A、B、C、D组,分别使用600 μmol/L的过氧化氢(H2O2)处理0、6、8、10 h,随后采用CCK-8法测定细胞活性以分析H2O2诱导氧化应激细胞模型的最佳干预时间。将HUVEC分为对照组(不做任何处理)、H2O2组(使用600 μmol/L的H2O2处理8 h以构建氧化应激细胞模型),采用q-PCR检测LncRNAMALAT1表达水平。将HUVEC分为对照组(不做任何处理)、H2O2组(使用600 μmol/L的H2O2处理8 h以构建氧化应激细胞模型)、H2O2+siRNA组(转染LncRNA MALAT1的siRNA后使用600 μmol/L的H2O2处理8 h以构建氧化应激细胞模型),采用Western blot法检测TLR4、MyD88、NF-κB表达水平。将HUVEC分为对照组(不做任何处理)、H2O2组(使用600 μmol/L的H2O2处理8 h以构建氧化应激细胞模型)、H2O2+siRNA组(转染LncRNA MALAT1的siRNA后使用600 μmol/L的H2O2处理8 h以构建氧化应激细胞模型),采用q-PCR检测TLR4、MyD88、NF-κB mRNA表达水平。结果 B、C、D组细胞活性均小于A组(P<0.05);C组细胞活性最接近60%,故H2O2诱导氧化应激细胞模型的最佳干预时间为8 h。H2O2组LncRNA MALAT1表达水平低于对照组(P<0.05)。H2O2+siRNA组TLR4、MyD88、NF-κB表达水平高于对照组(P<0.05);H2O2+siRNA组MyD88、NF-κB表达水平高于H2O2组(P<0.05)。H2O2组TLR4、MyD88mRNA表达水平高于对照组(P<0.05);H2O2+siRNA组TLR4、MyD88、NF-κB mRNA表达水平高于对照组、H2O2组(P<0.05)。结论 氧化应激环境下,LncRNA MALAT1表达水平降低,进而导致内皮细胞TLR4/MyD88/NF-κB信号通路被激活。
Abstract:
Objective To explore the effect of long chain non-coding RNA metastasis-associated lungadenocarcinoma tran 1 (LncRNA MALAT1) on Toll-like receptor 4 (TLR4) /myeloid differentiation factor 88 (MyD88) /nuclearfactor-kappa B (NF-κB) in endothelial cells under oxidative stress. Methods This experiment was conducted from Octoberto December 2022. Human umbilical vein endothelial cells (HUVECs) were divided into groups A, B, C and D, and treated with600 μmol/L hydrogen peroxide (H2O2) for 0, 6, 8 and 10 h, respectively. Then the cell activity was determined by CCK-8 methodto analyze the optimal intervention time of H2O2 to induce oxidative stress cell model. HUVECs were divided into control group(without any treatment) and H2O2 group (treated with 600 μmol/L H2O2 for 8 h to construct oxidative stress cell model) , and theexpression level of LncRNA MALAT1 was detected by q-PCR. HUVECs were divided into control group (without any treatment) ,H2O2 group (treated with 600 μmol/L H2O2 for 8 h to construct oxidative stress cell model) , H2O2+siRNA group (transfected withLncRNA MALAT1 siRNA and treated with 600 μmol/L H2O2 for 8 h to construct oxidative stress cell model) . The expressionlevels of TLR4, MyD88 and NF-κB were detected by Western blot. HUVECs were divided into control group (without anytreatment) , H2O2 group (treated with 600 μmol/L H2O2 for 8 h to construct oxidative stress cell model) , H2O2+siRNA group(transfected with LncRNA MALAT1 siRNA and treated with 600 μmol/L H2O2 for 8 h to construct oxidative stress cell model) . The mRNA expression levels of TLR4, MyD88 and NF-κB were detected by q-PCR. Results The cell activity of groups B, Cand D was lower than that of group A (P < 0.05) . The cell activity of group C was the closest to 60%, so the optimal interventiontime of H2O2 to induce oxidative stress cell model was 8 h. The expression level of LncRNA MALAT1 in H2O2 group was lowerthan that in control group (P < 0.05) . The expression levels of TLR4, MyD88 and NF-κB in H2O2+siRNA group were higher thanthose in control group (P < 0.05) . The expression levels of MyD88 and NF-κB in H2O2+siRNA group were higher than those inH2O2 group (P < 0.05) . The mRNA expression levels of TLR4 and MyD88 in H2O2 group were higher than those in control group(P < 0.05) . The mRNA expression levels of TLR4, MyD88 and NF-κB in H2O2+siRNA group were higher than those in controlgroup and H2O2 group (P < 0.05) . Conclusion Under oxidative stress, the expression level of LncRNA MALAT1 is decreased,which leads to the activation of TLR4/MyD88/NF-κB signaling pathway in endothelial cells.
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