肿瘤坏死因子-α在骨关节炎成纤维样滑膜细胞中上调β1，4-半乳糖基转移酶-I的表达

Tumor necrosis factor-α up-regulates the expression of β1, 4-Galactosyltransferase- I in fibroblast-like synoviocytes of osteoarthritis

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B1，禾GalT-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-timePCR）检测脂多糖和TNF-α刺激后，β1，4-GaIT-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-GalT．I（0．48±0．09），TNF-α（0．46±0．07）相比，骨关节炎滑膜组织中β1，4-GaIT-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，FLSs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-GalT- 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-GaiT- 1 and TNF-α were measured by reverse transcription-polymerase 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-GaiT- I and TNF-α was analyzed by double immunofluorescence. ANOVA and t-test was used for statistical analysis. Results ① Compared 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 131,4-GAIT-I and TNF-α; ② Lipopolysaccharide （LPS） could induce fibroblast-like synoviocytes （FLSs） β1,4-GAIT-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-GAIT-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-α to control β1,4-GaiT-I functions during inflammation in OA.