氧化应激环境中长链非编码RNA KLHL7-AS1对髓核细胞增殖和凋亡的影响及其机制  

Effects of long non-coding RNA KLHL7-AS1 on the proliferation and apoptosis of nucleus pulposus cells under the environment of oxidative stress and its mechanism

作　　者：杨钊琦 孙中仪 戴健[1] 马健 李瑶 孙宇 唐晓明[1] Yang Zhaoqi;Sun Zhongyi;Dai Jian;Ma Jian;Li Yao;Sun Yu;Tang Xiaoming(Department of Orthopedics,the First Hospital of Huaian City,Nanjing Medical University,Huaian 223300,China;Department of Orthopedics,Nanjing Jiangbei Hospital,Nanjing 210048,China)

机构地区：[1]南京医科大学附属淮安第一医院骨科,淮安223300 [2]南京江北医院骨科,南京210048

出　　处：《中华医学杂志》2025年第5期379-384,共6页National Medical Journal of China

基　　金：江苏省自然科学基金(BK20201130)。

摘　　要：目的研究氧化应激环境中长链非编码RNA KLHL7-AS1(LncRNA KLHL7-AS1)对髓核细胞增殖和凋亡的影响及其机制。方法应用人髓核细胞HUM-iCell-s012对细胞进行分组处理,未氧化的髓核细胞转染pcDNA空载体(pcDNA-control)以构建空白对照组(pcDNA-control)。参考以往氧化应激诱导椎间盘髓核细胞衰老的研究及前期实验,以400μmol/L的H_(2)O_(2)处理髓核细胞,建立氧化应激环境。氧化的髓核细胞分别转染pcDNA-control、KLHL7-AS1干扰RNA(siRNA-KLHL7-AS1)和KLHL7-AS1过表达质粒(pcDNA-KLHL7-AS1)以构建模型对照组(H_(2)O_(2)+pcDNA-control)、模型沉默组(H_(2)O_(2)+siRNA-KLHL7-AS1)以及模型过表达组(H_(2)O_(2)+pcDNA-KLHL7-AS1)。依上处理后,检测比较各组(n=3)KLHL7-AS1表达、细胞增殖及细胞凋亡情况,并检测比较各组细胞淋巴瘤基因-2蛋白(BCL-2)以及BCL-2相关蛋白(BAX)表达情况。结果模型对照组的KLHL7-AS1表达高于空白对照组(7.716±0.851比0.996±0.086),模型过表达组高于模型沉默组(19.592±1.747比4.222±0.039)(均P<0.001)。模型沉默组细胞活性与空白对照组相当(0.125±0.006比0.127±0.010,P=0.390),但细胞凋亡率较高(17.2%±1.7%比11.5%±1.0%),BCL-2蛋白表达亦较高(0.897±0.025比0.730±0.026),BAX蛋白表达较低(0.250±0.030比0.523±0.015)(均P<0.05)。模型沉默组的细胞活性高于模型对照组(0.125±0.006比0.076±0.008),细胞凋亡率较低(17.2%±1.7%比30.7%±2.3%),BCL-2蛋白表达较高(0.730±0.026比0.440±0.036),BAX蛋白表达较低(0.523±0.015比0.723±0.021),差异均有统计学意义(均P<0.001)。模型沉默组的细胞活性高于模型过表达组(0.125±0.006比0.039±0.006),细胞凋亡率较低(17.2%±1.7%比43.1%±1.9%),BCL-2蛋白表达较高(0.730±0.026比0.227±0.021),BAX蛋白表达较低(0.523±0.015比1.147±0.035),差异均有统计学意义(均P<0.001)。结论下调KLHL7-AS1的表达可促进髓核细胞的增殖,抑制髓核细胞的凋亡,其机制可能是通过调节蛋白BCL-2�Objective:To investigate the effects of long non-coding RNA KLHL7-AS1(LncRNA KLHL7-AS1)on the proliferation and apoptosis of nucleus pulposus cells under oxidative stress and its mechanisms.Methods:Human nucleus pulposus cells(HUM-iCell-s012)were divided into 4 groups,and unoxidized nucleus pulposus cells were transfected with an empty pcDNA vector(pcDNA-control)to serve as the blank control group.Based on previous studies on oxidative stress-induced nucleus pulposus cell senescence and preliminary experiments,oxidative stress was induced by treating nucleus pulposus cells with 400μmol/L H_(2)O_(2).Oxidized nucleus pulposus cells were transfected with pcDNA-control,KLHL7-AS1 interfering RNA(siRNA-KLHL7-AS1),or KLHL7-AS1 overexpression plasmid(pcDNA-KLHL7-AS1),forming the model control group(H_(2)O_(2)+pcDNA-control),the model silencing group(H_(2)O_(2)+siRNA-KLHL7-AS1),and the model overexpression group(H_(2)O_(2)+pcDNA-KLHL7-AS1),respectively.Following these treatments,KLHL7-AS1 expression levels,cell proliferation,cell apoptosis were measured and compared among the groups,and the expression levels of B-cell lymphoma 2(BCL-2)and BCL-2-associated X protein(BAX)in the groups of cells were compared too(n=3).Results:The expression of KLHL7-AS1 in the model control group was significantly higher than that in the blank control group(7.716±0.851 vs 0.996±0.086,P<0.001).Furthermore,the expression of KLHL7-AS1 in the model overexpression group was markedly higher than that in the model silencing group(19.592±1.747 vs 4.222±0.039,P<0.001).Compared to the blank control group,the model silencing group showed no significant change in cell viability(0.125±0.006 vs 0.127±0.010,P=0.390),an increased apoptosis rate(17.2%±1.7%vs 11.5%±1.0%,P<0.05),elevated BCL-2 protein expression(0.897±0.025 vs 0.730±0.026,P<0.05),and reduced BAX protein expression(0.250±0.030 vs 0.523±0.015,P<0.05).In comparison to the model control group,the model silencing group exhibited increased cell viability(0.125±0.006 vs 0.076±0.008),red

关 键 词：椎间盘 氧化应激 长链非编码RNA KLHL7-AS1 髓核细胞 

分 类 号：R73[医药卫生—肿瘤]