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[1]王培文,姚學(xué)東,俞海明,等.干擾NIX通過抑制自噬緩解脊髓神經(jīng)元的缺血再灌注損傷[J].福建醫(yī)藥雜志,2024,46(04):88-91.[doi:10.20148/j.fmj.2024.04.026]
　WANG Peiwen,YAO Xuedong,YU Haiming,et al.Interference of NIX alleviates spinal cord neurons ischemia-reperfusion injury by inhibiting autophagy[J].FUJIAN MEDICAL JOURNAL,2024,46(04):88-91.[doi:10.20148/j.fmj.2024.04.026]
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干擾NIX通過抑制自噬緩解脊髓神經(jīng)元的缺血再灌注損傷()
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《福建醫(yī)藥雜志》[ISSN:1002-2600/CN:35-1071/R]

卷:
46
期數(shù):
2024年04期
頁碼:
88-91
欄目:
基礎(chǔ)研究
出版日期:
2024-08-15

文章信息/Info

Title:
Interference of NIX alleviates spinal cord neurons ischemia-reperfusion injury by inhibiting autophagy
文章編號(hào):
1002-2600(2024)04-0088-04
作者:
王培文; 姚學(xué)東; 俞海明; 莊華烽; 許昊
福建醫(yī)科大學(xué)附屬第二醫(yī)院骨科, 泉州 362000
Author(s):
WANG Peiwen; YAO Xuedong; YU Haiming; ZHUANG Huafeng; XU Hao
Department of Orthopaedics,the Second Affiliated Hospital of Fujian Medical University,Quanzhou,Fujian 362000,China
關(guān)鍵詞:
NIX;  脊髓神經(jīng)元;  缺血再灌注;  自噬
Keywords:
NIX;  spinal cord neuron cells;  ischemia reperfusion;  autophagy
分類號(hào):
R394; R651.2
DOI:
10.20148/j.fmj.2024.04.026
文獻(xiàn)標(biāo)志碼:
B
摘要:
目的 觀察線粒體外膜蛋白NIX能否通過抑制自噬緩解脊髓神經(jīng)元缺血再灌注損傷。方法 利用H2O2(200 μmol/mL)處理分離的原代脊髓神經(jīng)元,以構(gòu)建體外缺血再灌注模型,通過轉(zhuǎn)染shNIX干擾NIX的表達(dá),并通過流式凋亡檢測(cè)與CCK-8增殖檢測(cè)觀察脊髓神經(jīng)元的凋亡情況,PCR檢測(cè)NIX基因的轉(zhuǎn)錄水平,蛋白免疫印跡法檢測(cè)NIX、LC3與Beclin蛋白的表達(dá)水平。結(jié)果 shNIX能顯著下調(diào)NIX基因的轉(zhuǎn)錄水平與蛋白的表達(dá)水平,抑制效果明顯,用于后續(xù)的實(shí)驗(yàn)操作。流式凋亡檢測(cè)與CCK-8增殖檢測(cè)表明,缺血再灌注損傷誘導(dǎo)脊髓神經(jīng)元出現(xiàn)凋亡,降低其增殖速度。缺血再灌注損傷促進(jìn)NIX、LC3和Beclin的表達(dá)。shNIX使缺血再灌注損傷誘導(dǎo)脊髓神經(jīng)元的凋亡減少,促進(jìn)了細(xì)胞的增殖,并且使LC3和Beclin的蛋白表達(dá)減少。結(jié)論 干擾NIX通過抑制自噬減弱了缺血再灌注誘導(dǎo)的脊髓神經(jīng)元損傷。
Abstract:
Objective This study aimed to observe whether interference of mitochondrial outer membrane protein NIX could alleviate spinal cord neurons ischemia-reperfusion injury by inhibiting autophagy. Methods Primary spinal cord neurons were isolated and treated with H2O2(200 μmol/mL)to construct the ischemia-reperfusion cell model in vitro.The expression of NIX was knocked down by transfection with shNIX. The apoptosis and proliferation of spinal cord neurons were observed by flow cytometry proliferation of CCK-8 proliferation detection. The transcriptional level of NIX gene was detected by PCR,and the protein expression of NIX,LC3,and Beclin were detected by Western blotting. Results shNIX significantly downregulated the transcription level and protein expression levels of NIX,and the inhibition effect is obvious,which is used for subsequent experimental operations. Flow cytometry proliferation of CCK-8 proliferation detection showed that ischemia-reperfusion injury induced apoptosis of spinal cord neurons with reduced proliferation rate. In addition,ischemia-reperfusion injury promoted the expressions of NIX,LC3,and Beclin. More importantly,shNIX reduced the apoptosis of spinal cord neurons,promoted cell proliferation,as well as reduced the protein expressions of LC3 and Beclin. Conclusion Interference of NIX attenuated ischemia-reperfusion induced spinal cord neurons injury by inhibiting autophagy.

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備注/Memo

備注/Memo:
基金項(xiàng)目:福建省自然科學(xué)基金項(xiàng)目(2020J01211)
更新日期/Last Update: 2024-08-15