文章摘要
李鑫,孙茂苍,崔山龙.微小 RNA-195降低寡聚态 β样淀粉蛋白 1-42诱导大鼠肾上腺嗜铬细胞炎症反应和凋亡作用机制[J].安徽医药,2022,26(1):106-111.
微小 RNA-195降低寡聚态 β样淀粉蛋白 1-42诱导大鼠肾上腺嗜铬细胞炎症反应和凋亡作用机制
The mechanism of miR-195 affects oligomeric AβAβ1-42 inducing inflammatory response and apoptosis in PC12 cells by regulating ATP13A2
  
DOI:10.3969/j.issn.1009-6469.2022.01.025
中文关键词: 微小 RNAs  阳离子转运蛋白质类  肾上腺嗜铬细胞  阿尔茨海默病  微小 RNA-195  抗炎  增殖  凋亡
英文关键词: MicroRNAs  Cation transport proteins  Adrenal chromaffin cells  Alzheimer disease  miR-195  Anti-inflammatory  Pro-liferation  Apoptosis
基金项目:
作者单位E-mail
李鑫 聊城市第四人民医院神经内科山东聊城 252000  
孙茂苍 聊城市第四人民医院神经内科山东聊城 252000  
崔山龙 聊城市第四人民医院神经内科山东聊城 252000 1545782053@qq.com 
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中文摘要:
      目的探讨微小 RNA-195(miR-195)对寡聚态 β样淀粉蛋白( Aβ)1-42诱导大鼠肾上腺嗜铬细胞(PC12细胞)炎症反应和凋亡的作用机制。方法本研究起止时间为 2018年 3—11月。 PC12细胞购自美国模式培养物保藏中心。运用 25 μmol/mL的 Aβ1-42处理 PC12细胞建立神经细胞损伤模型;将 PC12细胞分为 Aβ1-42+miR-con组、 Aβ1-42+miR-195组、 Aβ1-42+si-con组、 Aβ1-42+ATP酶阳离子转运 13A2小干扰 RNA(si-ATP13A2)组、 miR-con组、 miR-195组、 Aβ1-42+miR-195+pcDNA组、 Aβ1-42+miR-195+ATP13A2过表达质粒( pcDNA -ATP13A2)组。实时定量 PCR检测细胞中 miR-195、ATP13A2的 mRNA的表达; Western blotting检测细胞中 ATP13A2的蛋白表达;酶联免疫吸附实验检测细胞中白细胞介素 1β(IL-1β)、肿瘤坏死因子 α(TNF-α)的含量;噻唑蓝法、流式细胞术检测细胞的增殖和凋亡;双荧光素酶报告基因检测实验检测细胞中 miR-195与 ATP13A2的结合力。结果与对照组相比, Aβ1-42可显著下调 PC12细胞中 miR-195的表达[( 0.22±0.04)比( 1.01±0.06)]上调 ATP13A2的表达[(0.78±0.08)比( 0.41±0.05)];过表达 miR-195可促进 PC12细胞的增殖[(76.59±8.44)%比(52.73±5.82)%],,
英文摘要:
      Objective To investigatethe mechanism ofmiR-195 on the inflammatoryandapoptosis of PC12cellsinduced byoligomeric AβAβ1-42.Methods This research was conducted from March to November, 2018 PC12 cells were purchased from American Model Cul-ture Preservation Center. Nerve injury cell model was established by treatment of PC12 cells with 25 μmol/mL Aβ1-42; PC12 cells were di-videdinto Aβ1-42+miR-congroup,Aβ1-42+miR-195group,Aβ1-42+si-congroup,Aβ1-42+ATPase cationtransporting 13A2smallinter-feringRNA (si-ATP13A2)group,miR-con group,miR-195 group,Aβ1-42+miR-195+pcDNA group,Aβ1-42+miR-195+ATP13A2 overex-pressedplasmid(pcDNA-ATP13A2)group.ThemRNAexpressionofmiR-195andATP13A2weredetectedbyreal-timequantitativePCR. The protein expression of ATP13A2 was detected by Western blotting. The content of interleukin 1β(IL-1β) and tumor necrosis factor α (TNF-α)were detected by enzyme-linked immunosorbentassay.The cell proliferationand apoptosiswere detected by Methyl ThiazolylTet-razolium assay and cytometry; dual-luciferase reporter assay was used to detect the binding force of miR-195 and ATP13A2 in cells.Re. sults Compared with the control group, Aβ1-42 significantly down-regulated the expression of miR-195 (0.22±0.04) vs. (1.01±0.06) in PC12 cells and up-regulated the expression of ATP13A2 (0.78±0.08) vs. (0.41±0.05). Overexpression of miR-195 inhibited the expression of IL-1β [(875.28±118.47) pg/mL vs. (1753.84±215.92) pg/mL], TNF-α [(185.32±23.27) pg/mL vs. (268.75±27.67) pg/mL] and apoptosis [(17.76±1.87)% vs. (26.56±2.12) %],promotedproliferation [(76.59±8.44)% vs. (52.73±5.82)%] ofPC12cells;silencingATP13A2 inhib-ited the expression of IL-1β [(953.64±105.28) pg/mL vs. (1880.77±238.49) pg/mL], TNF-α [(152.81±22.46) pg/mL vs. (258.40±28.16) pg/ mL] and apoptosis [(18.03±1.82) % vs. (24.73±2.55) %], promoted proliferation [(78.59±9.76) % vs. (53.59±6.46) %] of PC12 cells. miR-195 inhibited the fluorescence activity of wild-type ATP13A2 cells and negatively regulated the expression of ATP13A2; overexpression of ATP13A2 reversed the effect of miR-195 on anti-inflammation, proliferation-promotion and apoptosis-inhibition of Aβ1-42-treated PC12 cells.Conclusion miR-195caninhibittheinflammatoryreactionandapoptosisofPC12cellsinducedbyAβ1-42.Themechanismisrelat-edtotargetingATP13A2,whichwillprovideanewtargetforthetreatmentofAlzheimer'sdisease.
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