苦参碱对椎间盘退变大鼠髓核细胞凋亡的影响  

Effect of matrine on apoptosis of nucleus pulposus cells in rats with intervertebral disc degeneration

作　　者：王冲 孔丽 崔学超 鲍铁周 Wang Chong;Kong Li;Cui Xuechao;Bao Tiezhou(College of Orthopedics and Traumatology,Henan University of Chinese Medicine,Zhengzhou 450000,Henan Province,China;Center for Neck,Shoulder,Lumbar and Leg Pain,Henan Luoyang Orthopedic Hospital(Henan Orthopedic Hospital),Luoyang 471002,Henan Province,China)

机构地区：[1]河南中医药大学骨伤学院,河南省郑州市450000 [2]河南省洛阳正骨医院(河南省骨科医院)颈肩腰腿痛中心,河南省洛阳市471002

出　　处：《中国组织工程研究》2024年第27期4281-4287,共7页Chinese Journal of Tissue Engineering Research

基　　金：河南省中医药科学研究专项项目(2023ZY2120),项目名称:温阳利水法在肩关节镜手术围手术期并发低体温患者中的应用,项目参与人:崔学超。

摘　　要：背景:髓核细胞凋亡是引发椎间盘退变的主要病理基础,炎症和过氧化反应是导致髓核细胞凋亡的重要因素。研究表明苦参碱具有抗氧化、衰老、炎症和细胞凋亡等作用,可作为治疗椎间盘退变的潜在药物。目的:观察苦参碱调节环状GMP-AMP合成酶-干扰素基因刺激因子(cGAS-STING)信号通路对椎间盘退变大鼠髓核细胞凋亡的影响。方法:①将处于对数生长期的大鼠髓核细胞分6组处理:除对照组外,模型组、苦参碱低剂量组、苦参碱高剂量组、空载组、苦参碱高剂量+cGAS过表达组采用氧糖剥夺建立椎间盘退变细胞模型,造模同时苦参碱低剂量组、苦参碱高剂量组分别给予0.4,0.8 mmol/L苦参碱处理,空载组转染空载质粒,苦参碱高剂量+cGAS过表达组给予0.8 mmol/L苦参碱处理并转染cGAS过表达质粒。处理24 h后,检测细胞活性与凋亡,细胞内活性氧、超氧化物歧化酶、肿瘤坏死因子α及白细胞介素1β水平,以及细胞内凋亡蛋白、cGAS-STING通路蛋白表达。②将60只SD大鼠随机分为6组,每组10只:除对照组外,模型组、苦参碱低剂量组、苦参碱高剂量组、空载组、苦参碱高剂量+cGAS过表达组建立椎间盘退变模型;造模12周后,苦参碱低剂量组、苦参碱高剂量组分别灌胃给予60,120 mg/kg苦参碱(1次/d),空载组尾静脉注射空载质粒(2次/周),苦参碱高剂量+cGAS过表达组灌胃给予120 mg/kg苦参碱+尾静脉注射cGAS过表达质粒,连续治疗3周。给药结束后取材,检测细胞凋亡,活性氧、超氧化物歧化酶、肿瘤坏死因子α及白细胞介素1β水平,以及凋亡蛋白、cGAS-STING通路蛋白表达。结果与结论:①与对照组比较,模型组细胞活性、超氧化物歧化酶水平降低(P<0.05),细胞凋亡率、活性氧含量、肿瘤坏死因子α和白细胞介素1β水平及cGAS、STING、cleaved caspase-3和Bax蛋白表达升高(P<0.05);苦参碱可呈剂量依赖性地改善上述因细胞�BACKGROUND:Nucleus pulposus cell apoptosis is the main pathological basis for intervertebral disc degeneration,and inflammation and peroxidation are important factors leading to apoptosis in the nucleus pulposus.Studies have shown that matrine has antioxidant,senescent,inflammatory and apoptotic effects,and may be a potential drug for the treatment of disc degeneration.OBJECTIVE:To investigate the effect of matrine on apoptosis of nucleus pulposus cells in rats with intervertebral disc degeneration by regulating the cyclic GMP-AMP synthase(cGAS)-stimulator of interferon genes(STING)signaling pathway.METHODS:(1)Nucleus pulposus cells of rats at a logarithmic phase were randomly separated into a control group,a model group,a low-dose matrine group,a high-dose matrine group,an empty group,and a high-dose matrine+cGAS overexpression group.Except for the control group,cell models of intervertebral disc degeneration were established in the other groups through oxygen-glucose deprivation.At the same time of modeling,the low-dose and high-dose groups were treated with 0.4 and 0.8 mmol/L matrine,respectively,and the empty group was transfected with the empty plasmid,while the high-dose+cGAS overexpression group was treated with 0.8 mmol/L matrine with the transfection of the cGAS overexpression plasmid.After 24 hours of treatment,cell activity and apoptosis,intracellular levels of reactive oxygen species,superoxide dismutase,tumor necrosis factor α and interleukin 1β,and intracellular expression of apoptotic proteins and cGAS-STING pathway proteins were detected.(2)Sixty Sprague-Dawley rats were randomized into six groups(n=10 per group):control group,model group,low-dose matrine group,high-dose matrine group,empty group,and high-dose+cGAS overexpression group.After 12 weeks of modeling,60 and 120 mg/kg matrine were given by gavage in the low-dose and high-dose matrine groups,respectively(once a day),and the empty plasmid was injected into the tail vein in the empty group(2 times/week),while the high-dose+cGAS overexpre

关 键 词：苦参碱 cGAS-STING通路 椎间盘退变 髓核细胞 细胞凋亡 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R285.5