青藤碱可有效抑制白细胞介素1β介导的髓核细胞凋亡  

Sinomenine effectively inhibits interleukin-1beta-induced apoptosis in nucleus pulposus cells

作　　者：王倩[1] 卢子昂 李利和 吕超亮[1] 王盟[3] 张存鑫 Wang Qian;Lu Ziang;Li Lihe;Lyu Chaoliang;Wang Meng;Zhang Cunxin(Department of Spine Surgery,Jining No.1 People’s Hospital,Jining 272000,Shandong Province,China;School of Clinical Medicine,Jining Medical University,Jining 272067,Shandong Province,China;Clinical Medical Experiment Center,Jining No.1 People’s Hospital,Jining 272000,Shandong Province,China)

机构地区：[1]济宁市第一人民医院脊柱外科,山东省济宁市272000 [2]济宁医学院,临床医学院,山东省济宁市272067 [3]济宁市第一人民医院临床医学实验中心,山东省济宁市272000

出　　处：《中国组织工程研究》2024年第2期224-230,共7页Chinese Journal of Tissue Engineering Research

基　　金：山东省中医药科技项目(Q-2022025),项目负责人:张存鑫;济宁市重点研发计划项目(2020JKNS008),项目负责人:张存鑫;济宁市重点研发计划项目(2019SMNS003),项目负责人:王盟。

摘　　要：背景:椎间盘退变是导致脊柱退行性疾病的基础,然而目前尚无有效的治疗药物。目的:探讨青藤碱是否可以抑制白细胞介素1β诱导的髓核细胞凋亡及其分子机制。方法:采用胰酶联合Ⅱ型胶原酶消化法体外培养大鼠髓核细胞,并绘制细胞生长曲线,采用CCK-8法筛选合适的青藤碱药物浓度。将髓核细胞分为对照组、青藤碱组、白细胞介素1β组、青藤碱+白细胞介素1β组、锌原卟啉(血红素氧合酶1抑制剂)组、锌原卟啉+青藤碱组、锌原卟啉+白细胞介素1β组、青藤碱+锌原卟啉+白细胞介素1β组。分别检测各组髓核细胞增殖活性、活性氧含量、凋亡率及血红素氧合酶1的表达情况。结果与结论:①体外培养的大鼠髓核细胞呈现多角形、三角形、短楔形等形态,其呈现“S”型曲线生长,接种第1-3天生长缓慢,第4-6天生长迅速,第七八天生长速度缓慢,进入“平台期”,细胞数量不再增加;②当青藤碱的浓度≤80μmol/L时,髓核细胞的增殖活性不会受到显著影响(P>0.05);③白细胞介素1β可以显著降低髓核细胞的增殖活性,增加活性氧含量,导致细胞凋亡(P<0.01);④当采用青藤碱干预后,不仅可以促进血红素氧合酶1的表达(P<0.05),而且可以抑制白细胞介素1β诱导的髓核细胞增殖活性降低、活性氧含量和凋亡率增加(P<0.05),其作用可被锌原卟啉逆转(P<0.01)。BACKGROUND:Intervertebral disc degeneration is the basis of spinal degenerative diseases;however,there is no effective treatment.OBJECTIVE:To investigate whether sinomenine can inhibit interleukin-1β-induced apoptosis in nucleus pulposus cells and its molecular mechanism.METHODS:Rat nucleus pulposus cells were cultured in vitro by trypsin combined with type II collagenase digestion,and the cell growth curve was plotted.An appropriate sinomenine concentration was determined using the cell counting kit-8 kit.Nucleus pulposus cells were divided into control group,sinomenine group,interleukin-1βgroup,sinomenine+interleukin-1βgroup,zinc protoporphyrin group,zinc protoporphyrin+sinomenine group,zinc protoporphyrin+interleukin-1βgroup,and sinomenine+zinc protoporphyrin+interleukin-1βgroup.Proliferative activity,reactive oxygen species content,apoptosis rate,and heme oxygenase-1 expression in nucleus pulposus cells were detected.RESULTS AND CONCLUSION:The rat nucleus pulposus cells cultured in vitro were polygonal,triangular,and short wedge-shaped,and the cell growth showed an“S”curve.The cells grew slowly in the first 3 days of culture,rapidly in 4-6 days,and slowly again in 7-8 days.The cells then entered the“platform stage”where the number of cells no longer increased.The proliferative activity of myeloid cells showed no significant changes when the concentration of sinomenine was≤80μmol/L(P>0.05).Interleukin-1βsignificantly reduced the proliferative activity of nucleus pulposus cells,increased the content of reactive oxygen species and led to apoptosis(P<0.01).Sinomenine intervention not only promoted heme oxygenase-1 expression(P<0.05)but also inhibited interleukin-1β-induced decrease in proliferative activity and increase in reactive oxygen species content and apoptosis rate in nucleus pulposus cells(P<0.05).These effects could be reversed by zinc protoporphyrin(P<0.01).

关 键 词：青藤碱 白细胞介素1Β 血红素氧合酶1 髓核细胞 细胞增殖 细胞凋亡 活性氧 椎间盘 椎间盘退变 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R681.57