壳多糖酶3样蛋白质1在腰椎关节突关节退行性病变中的作用  

Chitinase-3-like protein 1 plays a role in lumbar facet joint degeneration

作　　者：陈元振 王磊 谢良玉 曹盛楠 于秀淳 李华忠 黄伟敏 师彬 王丹丹 Chen Yuanzhen;Wang Lei;Xie Liangyu;Cao Shengnan;Yu Xiuchun;Li Huazhong;Huang Weimin;Shi Bin;Wang Dandan(Department of Orthopaedics,Neck-Shoulder and Lumbocrural Pain Hospital of Shandong First Medical University,Bone Biomechanics Engineering Laboratory of Shandong Province,Jinan 250062,China;Department of Orthopaedics,the 960th Hospital of the People's Liberation Army,Jinan 250031,China;Shandong First Medical University&Shandong Academy of Medical Sciences,Jinan 250117,China)

机构地区：[1]山东第一医科大学附属颈肩腰腿痛医院(山东省骨生物力学工程实验室)骨科,济南250062 [2]解放军第九六⚪医院骨科,济南250031 [3]山东第一医科大学(山东省医学科学院),济南250117

出　　处：《中华骨科杂志》2022年第17期1148-1155,共8页Chinese Journal of Orthopaedics

基　　金：泰山学者特聘专家计划(ts201511074);山东省自然科学基金面上项目(ZR2019MH134);山东第一医科大学学术提升计划(2019QL003)。

摘　　要：目的探讨壳多糖酶3样蛋白质1(chitinase-3-like protein 1,CHI3L1)对软骨细胞生物学功能的影响在关节突关节退行性病变中的作用。方法收集人关节突关节软骨组织标本,检测CHI3L1基因的mRNA相对表达量,分析关节突关节退变等级与患者性别、年龄、CHI3L1基因mRNA相对表达量的相关性。体外培养人软骨细胞,检测CHI3L1组与对照组软骨细胞增殖、周期、凋亡和相关炎症因子表达的差异,以信号传导通路蛋白芯片分析CHI3L1调节软骨细胞退变的作用机制。结果关节突关节退变等级与CHI3L1基因mRNA相对表达量呈正相关(r=0.76,P<0.001),与性别不相关(r=-0.12,P=0.500),与年龄呈正相关(r=0.47,P=0.005)。退变组CHI3L1基因mRNA表达量升高,且随退变等级加重表达量升高(F=18.90,P<0.001)。与对照组比较,CHI3L1组软骨细胞在培养48 h(OD_(490)/fold=7.132)、72 h(OD_(490)/fold=4.803)、96 h(OD_(490)/fold=2.431)和120 h(OD_(490)/fold=0.009)的增殖倍数显著降低。CHI3L1组软骨细胞处于G1期、S期和G2/M期的比例分别为85.03%±3.05%、12.78%±2.29%和0.90%±0.76%,对照组分别为73.93%±2.73%、22.81%±1.93%和0.99%±0.87%;两组软骨细胞G1期比例(t=4.70,P<0.001)和S期比例(t=5.80,P<0.001)的差异均有统计学意义;两组软骨细胞凋亡百分比分别为8.64%±0.76%和5.68%±1.13%,差异有统计学意义(t=4.47,P<0.001);CHI3L1组软骨细胞IL-6的表达量为(49.60±0.01)pg/ml,大于对照组的(47.88±0.01)pg/ml(t=132.72,P<0.001),TNF-α的表达量为(95.93±0.02)pg/ml,大于对照组的(90.69±0.02)pg/ml(t=376.10,P<0.001)。蛋白芯片检测到表达量显著差异的蛋白共53个,蛋白磷酸化水平显著差异的蛋白43个。通过生物信息学分析筛选出差异蛋白可能参与的信号通路共16个。MAPK信号通路中,CHI3L1组软骨细胞MAPK1和RAF1蛋白的表达量(分别为1.094±0.00、0.988±0.00)较对照组(分别为0.814±0.00、0.786±0.00)升高(t=103.16,P<0.001;t=54.32,P<0.001)。结论CHI3L1的表�Objective To investigate the effect of CHI3L1 on the biological function of chondrocytes and its role in lumbar facet joint degeneration.Methods The human lumbar facet joint articular cartilage were collected,and the relative mRNA expression of CHI3L1 gene detected by quantitative fluorescence PCR.Then explored the correlation between joint degeneration and gender,age and relative mRNA expression of CHI3L1.Human chondrocytes were cultured in vitro.The effects of CHI3L1 on chondrocyte proliferation,cycling,and apoptosis,as well as expression of related inflammatory factors,were investigated.The mechanism by which CHI3L1 regulates the degeneration of articular cartilage was investigated using the signal transduction pathway protein chip.Results There was a positive correlation between the grade of degeneration in lumbar facet joint and the relative expression of CHI3L1 gene mRNA(r=0.76,P<0.001).There was no correlation with the patient's gender(r=-0.12,P=0.500).A positive correlation between the age of patients and the relative expression of CHI3L1 gene mRNA was found(r=0.47,P=0.005).Compared with the non-degenerative group,the expression of CHI3L1 gene mRNA significantly increased in the degenerative group,and the expression of CHI3L1 gradually increased with the aggravation in the grade of degeneration(F=18.90,P<0.001).Compared with the non-degenerative group,the chondrocytes in the CHI3L1 group had significantly lower proliferation at 48 h(OD_(490)/fold=7.132),72 h(OD_(490)/fold=4.803),96 h(OD_(490)/fold=2.431)and 120 h(OD_(490)/fold=0.009).The ratio of chondrocytes in G1 phase,S phase and G2/M phase were 85.03%±3.05%,12.78%±2.29%and 0.90%±0.76%in the CHI3L1 group,and 73.93%±2.73%,22.81%±1.93%and 0.99%±0.87%in control group,respectively.There were significant differences in the percentage of chondrocytes in G1 phase(t=4.70,P<0.001)and S phase(t=5.80,P<0.001)between the two groups.The percentages of apoptosis in chondrocyte in CHI3L1 group and control group were 8.64%±0.76%and 5.68%±1.13%,which has a stat

关 键 词：椎关节突关节 骨关节炎 软骨细胞 壳多糖酶3样蛋白质1 

分 类 号：R687.4[医药卫生—骨科学]