摘要
目的观察不同浓度巨噬细胞抑制因子-1(MIC-1)对老年膝OA患者软骨细胞增殖代谢的影响。方法收集健康年轻人和老年膝OA患者的血清、膝关节软骨标本各10例,检测标本中MIC-1的含量变化。常规培养人软骨细胞,对照组单纯加培养液,实验组培养液中含不同浓度重组人MIC-1蛋白,其中,实验组1浓度为10 ng/ml,实验组2浓度为100 ng/ml和实验组3浓度为200 ng/ml,培养48 h后,5-乙炔基-2-脱氧尿苷(EdU)检测软骨细胞增殖情况,蛋白质印迹法和实时荧光定量(qRT)-PCR检测软骨细胞pSmad2、Smad2和Col2等指标的表达情况。统计学分析采用t检验,单因素方差分析(ANOVA),Tukey法。结果年轻人血清中MIC-1蛋白相对表达量(0.89±0.11)高于老年人(0.48±0.12),差异有统计学意义(t=2.58,P<0.05)。对照组EdU阳性细胞百分比为(26.8±1.5)%,实验组1为(26.2±1.6)%,实验组2为(35.2±1.8)%,实验组3为(45.6±2.7)%,多组间比较差异有统计学意义(F=21.45,P<0.01)。pSmad2蛋白相对表达量在多组之间比较差异有统计学意义(F=18.32,P<0.01),其中,实验组2(1.25±0.11)和实验组3(0.93±0.22)分别高于对照组(0.54±0.10);多组比较Col2蛋白相对表达量差异有统计学意义(F=17.90,P<0.01),其中,实验组2(1.28±0.14)和实验组3(1.74±0.14)分别高于对照组(0.80±0.04)。Col2的mRNA相对表达量在多组间比较差异有统计学意义(F=17.90,P<0.01),其中,实验组2(1.13±0.16)和实验组3(1.65±0.15)分别高于对照组(0.49±0.10)。结论老年膝OA患者血清和软骨组织MIC-1含量低于年轻人,MIC-1可以促进老年膝关节软骨细胞增殖。
Objective To observe the effect of different concentrations of macrophage inhibitory cytokine(MIC)-1 on proliferation of old chondrocytes.Methods The serum and the cartilage specimens of the young and the old were collected respectively,and the content of MIC-1 in serum and cartilage was detected.Human chondrocytes were cultured,the control group was treated with culture medium alone,and the culture medium of the experimental group contained recombinant human MIC-1 protein at different concentrations,including 10 ng/ml in experimental group 1,100 ng/ml in experimental group 2 and 200 ng/ml in experimental group 3.After 48 h of culture,the proliferation of chondrocytes was detected by EdU,and the expression of pSmad2 Smad2 and Col2 in chondrocytes was detected by Western blot and Quantitative real time polymerase chain reaction(qRT-PCR).T test was used for comparison between the two groups.The single factor analysis of variance(ANOVA)and Tukey method were used for the multi-group comparison.Results The relative expression level of MIC-1 protein in serum of young people(0.89±0.11)was higher than that of old people(0.48±0.12),and the difference was statistically significant(t=2.58,P<0.05).The percentage of EdU positive cells in the control group was(26.8±1.5)%,that in the experimental group 1 was(26.2±1.6)%,that in the experimental group 2 was(35.2±1.8)%,and that in the experimental group 3 was(45.6±2.7)%.The difference between groups was statistically significant(F=21.45,P<0.01).The relative expression level of pSmad2 protein was significantly different between groups(F=18.32,P<0.01),among which,experimental group 2(1.25±0.11)and experimental group 3(0.93±0.22)were higher than control group(0.54±0.10),respectively.There were statistically significant differences in Col2 protein relative expression between groups(F=17.90,P<0.01),with experimental group 2(1.28±0.14)and experimental group 3(1.74±0.14)being higher than control group(0.80±0.04),respectively.The relative expression level of Col2 mRNA was significantly different between groups(F=17.90,P<0.01),with experimental group 2(1.13±0.16)and experimental group 3(1.65±0.15)being higher than control group(0.49±0.10).Conclusion The MIC-1 content of the serum and the cartilage in young people is higher than that in old people,and MIC-1 can promote the proliferation of chondrocytes in old.
作者
王东明
翟旭雯
耿翔
李鹏翠
卫小春
李璐
Wang Dongming;Zhai Xuwen;Geng Xiang;Li Pengcui;Wei Xiaochun;Li Lu(Department of Orthopedics,the Second Hospital of Shanxi Medical University,Shanxi Key Laboratory of Bone and Soft Tissue Injury Repair,Taiyuan 030001,China;the Simulation Hospital of Shanxi Medical University,Jinzhong 030619,China;Department of Pharmacy,Shanxi Health Vocational College,Jinzhong 030619,China)
出处
《中华风湿病学杂志》
CAS
CSCD
北大核心
2019年第12期839-844,I0004,共7页
Chinese Journal of Rheumatology
