摘要
探讨山柰酚(kaempferol,KAE)通过p38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)信号通路对椎间盘退变(intervertebral disc degeneration,IDD)的作用机制。将大鼠随机均分为空白组、模型组、KAE低剂量组、KAE中剂量组和KAE高剂量组,针刺尾椎椎间盘造模,术后4周开始灌胃给药,连续给药8周后行磁共振成像检查并取材;体外构建肿瘤坏死因子-α(tumor necrosis factor-alpha,TNF-α)诱导的髓核细胞(nucleus pulposus cells,NPCs)炎症模型,使用anisomycin激动p38 MAPK信号通路。将NPCs分为空白组、模型组、KAE组、激动剂组、KAE+激动剂组,NPCs按照分组分别处理1 d后,CCK-8试剂检测细胞增殖活性,蛋白免疫印迹法检测细胞相关蛋白表达水平,实时荧光定量聚合酶链式反应实验检测细胞相关mRNA表达,TUNEL染色检测骨细胞凋亡情况,免疫荧光染色检测Ⅱ型胶原、基质金属蛋白酶3(matrix metalloproteinase 3,MMP3)。体内实验结果显示,治疗组大鼠IDD程度较于模型组有明显改善,其中KAE中剂量组治疗效果明显优于KAE低、高剂量组。体外实验结果显示,KAE治疗可以显著提高NPCs增殖活性,下调NPCs中Bcl-2相关X蛋白、白细胞介素-6、白细胞介素-17A、MMP3、血小板反应蛋白基序解聚蛋白聚糖酶5蛋白表达水平并抑制p38 MAPK信号通路相关蛋白磷酸化水平;anisomycin激动p38 MAPK信号通路后,KAE治疗作用下降。综上研究表明,KAE能改善退变的NPCs细胞增殖活性,降低炎症水平,延缓大鼠IDD进展,其机制与调控p38 MAPK信号通路有关。
This study investigated the mechanism by which kaempferol(KAE)affected intervertebral disc degeneration(IDD)through the p38 mitogen-activated protein kinase(p38 MAPK)signaling pathway.Rats were randomly divided into five groups:control group,model group,low-dose KAE group,medium-dose KAE group,and high-dose KAE group.An IDD model was established by needle puncture of the caudal intervertebral discs.Four weeks post-surgery,the rats were administered KAE via gavage for 8 consecutive weeks.Magnetic resonance imaging(MRI)was performed,and samples were collected.In vitro,an inflammation model of nucleus pulposus cells(NPCs)induced by tumor necrosis factor-alpha(TNF-α)was constructed.Anisomycin was used to activate the p38 MAPK signaling pathway.NPCs were divided into blank,model,KAE,agonist,and KAE+agonist groups.After 1 day of treatment,cell proliferation activity was assessed using the CCK-8.Protein expression levels were determined by Western blot,and mRNA expression was measured by real-time quantitative polymerase chain reaction.Cell apoptosis was detected by TUNEL staining,and immunofluorescence staining was used to detect typeⅡcollagen and matrix metalloproteinase 3(MMP3).In vivo results indicated significant improvement in the degree of IDD in the treatment groups compared to the model group,with the medium-dose group showing more pronounced therapeutic effects than the low-and high-dose groups.In vitro results demonstrated that KAE treatment significantly enhanced NPC proliferation activity,down-regulated the expression levels of Bcl-2-associated X protein(Bax),interleukin-6(IL-6),interleukin-17A(IL-17A),MMP3,and a disintegrin and metalloproteinase with thrombospondin motifs 5,and inhibited the phosphorylation of p38 MAPK pathway-related proteins.Activation of the p38 MAPK signaling pathway by anisomycin reduced the therapeutic effects of KAE.The study concluded that KAE could improve the proliferation activity of degenerated NPCs,reduce inflammation levels,and slow the progression of IDD in rats,and the m
作者
汪陈莫及
吴亚东
梁松林
高尚
岳泽林
孔鹿鸣
高宽惠
李念虎
WANG Chen-mo-ji;WU Ya-dong;LIANG Song-lin;GAO Shang;YUE Ze-lin;KONG Lu-ming;GAO Kuan-hui;LI Nian-hu(the First Clinical Medical College of Shandong University of Traditional Chinese Medicine,Ji'nan 250014,China;Rizhao City Hospital of Traditional Chinese Medicine,Rizhao 276800,China;Afiliated Hospital of Shandong University of Traditional Chinese Medicine,Ji'nan 250014,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2024年第21期5721-5729,共9页
China Journal of Chinese Materia Medica
基金
山东省自然科学基金创新发展联合基金项目(ZR2021LZY006)
山东省高等学校科技计划项目(J17KA242)。
关键词
山柰酚
p38
MAPK信号通路
椎间盘退变
髓核细胞
椎间盘
kaempferol
p38 MAPK signaling pathway
intervertebral disc degeneration
nucleus pulposus cells
intervertebral disc
