甲基莲心碱可抑制白细胞介素1β诱导的软骨细胞凋亡  

Neferine inhibits interleukin-1beta-induced chondrocyte apoptosis

作　　者：周观金 李亚楠 李涛 Zhou Guanjin;Li Yanan;Li Tao(Department of Orthopedics,Wuhan NO.4 Hospital,Wuhan 430030,Hubei Province,China)

机构地区：[1]武汉市第四医院骨科,湖北省武汉市430030

出　　处：《中国组织工程研究》2024年第35期5624-5629,共6页Chinese Journal of Tissue Engineering Research

基　　金：武汉市中医药科研项目(WZ22Q20),项目负责人:李亚楠。

摘　　要：背景:细胞凋亡参与了关节退行性疾病的形成过程,因此抗软骨细胞凋亡可能是治疗骨关节炎的有效途径。甲基莲心碱具有抗炎、抗肿瘤、抗凋亡等广泛的药理活性,其对软骨细胞凋亡的影响尚不明确。目的:探讨甲基莲心碱对白细胞介素1β诱导的软骨细胞凋亡的影响,阐明可能的作用机制。方法:(1)取对数生长期大鼠软骨细胞,分5组干预:对照组常规培养,模型组加入白细胞介素1β处理2 h后常规培养24 h,低、中、高浓度药物组加入白细胞介素1β处理2 h后分别加入5,10,20μmol/L甲基莲心碱培养24 h。培养结束后,检测细胞凋亡、细胞上清中软骨糖蛋白39及Ⅱ型胶原水平、凋亡相关蛋白的m RNA与蛋白表达、PERK/ATF4信号通路相关蛋白的m RNA与蛋白表达。(2)取对数生长期大鼠软骨细胞,分4组干预:对照组与模型组干预同上,药物组加入白细胞介素1β处理2 h后再加入20μmol/L甲基莲心碱培养24 h,激活剂组加入白细胞介素1β处理2 h后再加入20μmol/L甲基莲心碱、PERK/ATF4信号通路激活剂CCT020312培养24 h。培养结束后,采用CCK-8法检测细胞增殖,流式细胞术检测细胞凋亡,凋亡相关蛋白与PERK/ATF4信号通路相关蛋白的m RNA与蛋白表达。结果与结论:(1)与对照组比较,模型组细胞凋亡率增加(P<0.05)、增殖活性降低(P<0.05),软骨糖蛋白39水平升高(P<0.05),Ⅱ型胶原水平降低(P<0.05),Bcl-2蛋白的m RNA与蛋白表达降低(P<0.05),Bax蛋白的m RNA与蛋白表达、caspase-3 m RNA表达、Cleaved-caspase-3蛋白表达均升高(P<0.05),PERK、ATF4、CHOP m RNA表达升高(P<0.05),p-PERK、ATF4、CHOP蛋白表达升高(P<0.05);与模型组比,甲基莲心碱可逆转白细胞介素1β对软骨细胞的上述影响,并且呈现浓度依赖性;(2)与药物组比较,激活剂组细胞凋亡率增加(P<0.05)、增殖活性降低(P<0.05),caspase-3、ATF4、CHOP m RNA表达升高(P<0.05),Cleaved-caspase-3、ATF4、CHOP蛋白表达均升�BACKGROUND:Apoptosis is involved in the formation of degenerative joint diseases.Therefore,anti-chondrocyte apoptosis may be an effective way to treat osteoarthritis.Neferine has a wide range of pharmacological activities including anti-inflammatory,anti-tumor and anti-apoptotic activities,and its effect on chondrocyte apoptosis is not clear.OBJECTIVE:To investigate the effect of neferine on chondrocyte apoptosis induced by interleukin-1βand elucidate its possible mechanism.METHODS:(1)The rat chondrocytes at logarithmical stage were taken and intervened in five groups.The control group was cultured routinely.The model group was routinely cultured for 24 hours after treatment with interleukin-1βfor 2 hours.The low-,medium-,and high-dose groups were treated with interleukin-1βfor 2 hours and then cultured with 5,10,and 20μmol/L neferine for 24 hours,respectively.At the end of culture,cell apoptosis,chondroglycoprotein 39 and type II collagen levels in cell supernatants,mRNA and protein expression of apoptosis-related proteins,and mRNA and protein expression of proteins related to the protein kinase R-like endoplasmic reticulum kinase(PERK)/activating transcription factor 4(ATF4)signaling pathway were detected.(2)Rat chondrocytes at logarithmic growth period were taken and divided into four groups:control group and model group were treated with the same intervention as above,drug group was treated with interleukin-1βfor 2 hours and then cultured with 20μmol/L neferine for 24 hours,and activator group was treated with interleukin-1βfor 2 hours and then cultured with 20μmol/L neferine and CCT020312,an activator of PERK/ATF4 signaling pathway,for 24 hours.At the end of culture,cell proliferation was detected by cell counting kit-8 assay,apoptosis was detected by flow cytometry,and mRNA and protein expressions of apoptosis-related proteins and PERK/ATF4 signaling pathway-related proteins were detected.RESULTS AND CONCLUSION:(1)Compared with the control group,the model group showed increased apoptosis(P<0.05),dec

关 键 词：甲基莲心碱 白细胞介素1Β 软骨细胞 细胞凋亡 内质网 PERK/ATF4信号通路 

分 类 号：R459.9[医药卫生—治疗学] R319[医药卫生—临床医学] R285.5