目的 探讨肿瘤坏死因子(TNF)-α与β1,4-半乳糖基转移酶-Ⅰ(β1,4-GalT-Ⅰ)在骨关节炎滑膜炎症过程中的相互关系.方法 提取各8例骨关节炎患者(试验组)和膝关节游离体患者(对照组)的滑膜组织,运用反转录-聚合酶链反应(RT-PCR)检测滑膜组织中β1,4-GaIT-Ⅰ和TNF-α表达水平,运用免疫荧光双标法检测β1,4-GalT-Ⅰ和TNF-α在滑膜组织中的共定位情况；培养成纤维样滑膜细胞(FLSs),运用酶联免疫吸附试验(ELISA)法检测脂多糖刺激后TNF-α表达变化及运用实时荧光定量聚合酶链反应(real-time PCR)检测脂多糖和TNF-α刺激后,β1,4-GalT-Ⅰ的表达变化；运用t检验和单因素方差分析进行统计学分析.结果 ①与对照组[β1,4-GaiT-Ⅰ(0.48±0.09),TNF-α(0.46±0.07)相比,骨关节炎滑膜组织中β1,4-GalT-Ⅰ(0.94±0.16)和TNF-α(1.19±0.19)表达均明显增加(t=3.47,t=4.06,均P＜0.01),且存在共定位；②脂多糖能够诱导FLSs β1,4-GalT-Ⅰ[11.2±0.9与2.9±0.5(量效),22.3±2.3与4.4±0.9(时效),F=83.03,F=157.58,均P＜0.05]表达增加；③脂多糖在FLSs中促进TNF-α[(1256±96)pg/ml与(101±7)pg/ml,F=124,F=93.6,均P＜0.01]表达增加；③TNF-α在FLSs中也诱导β1,4-GalT-Ⅰ[23.2±1.9与8.4±1.3(量效),23.9±1.8与11.5±1.3(时效),F=431.96,均P＜0.05]的表达增加.结论 在骨关节炎的滑膜炎症过程中,HLSs可能运用TNF-α调控β1,4-GalT-Ⅰ的表达.%Objective To analyze whether β1,4-galactosyltransferase-Ⅰ(β1,4-GaiT-Ⅰ)expression correlates with the expression of tumor necrosis factor(TNF)-α in osteoarthritis(OA).Methods Synovial tissue samples from eight OA patients and eight healthy people were obtained as the experimental group and controls respectively.The mRNA levels of β1,4-GalT-Ⅰ and TNF-α were measured by reverse transcriptionpolymerase chain reaction(RT-PCR)and real-time PCR.Enzyme linked immunosorbent assay(ELISA)was used to test the expression of TNF-α in the protein level.Cellular colocalization of β1,4-GalT-Ⅰ and TNF-α was analyzed by double immunofluorescence.ANOVA and t-test was used for statistical analysis.Results ①Compared with the control group[β1,4-GalT-Ⅰ(0.48±0.09),TNF-α(0.46±0.07)],the expression of β1,4-GalT-Ⅰ(0.94±0.16)and TNF-α(1.19±0.19)were significantly increased in OA synovial tissue(t=3.47,t=4.06,P＜0.01)and there was colocalization between β1,4-GalT-Ⅰ and TNF-α;② Lipopolysaccharide (LPS)could induce fibroblast-like synoviocytes(FLSs)β1,4-GalT-Ⅰ[11.2±0.9 vs 2.9±0.5(dose effect),22.3±2.3 vs 4.4±0.9(time effect),F=83.03,F=157.58,P＜0.05]overexpression;③ LPS could induce FLSs TNF-α[(1256±96)vs(101±7)pg/ml,F=431.96,P＜0.01]overexpression;④ Not only endogenous TNF-α,but exogenous TNF-α could induce FLSs β1,4-GalT-Ⅰ[23.2±1.9 vs 8.4±1.3(dose effect),23.9±1.8 vs 11.5±1.3(time effect),F=124,F=93.6,P＜0.05]overexpression.Conclusion It is possible that FLSs mayuse TNF-αto control β1,4-GalT-Ⅰ functions during inflammation in OA.
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