山梨醇增加糖尿病患者腰椎管狭窄症炎性反应因子的表达

Expression of inflammatory factors are increased by sorbitol in lumbar spinal stenosis of diabetic patients

目的探讨糖尿病腰椎管狭窄患者黄韧带增生肥厚的发生机制。方法 24例糖尿病和20例非糖尿病的腰椎管狭窄患者列为研究对象,观测黄韧带标本结构,D-Sorbitol/Xylitol试剂盒检测山梨醇水平。体外实验中使用小鼠成纤维细胞(NIH3T3)细胞系,用Western blot及q PCR分别检测高糖培养条件及醛糖还原酶抑制剂(ARI):依帕司他(EP)作用对细胞炎性反应因子及TGF-β表达水平的影响。结果糖尿病组较非糖尿病组的山梨醇水平更高、黄韧带平均厚度更大、标本弹力纤维降解、胶原纤维增生更为显著、免疫组化CD68阳性染色率更高(P<0.01);体外实验中,NIH3T3细胞系在高糖培养与正常糖浓度培养相比山梨醇、促炎性细胞因子和TGF-β表达水平更高,而山梨醇、促炎性细胞因子和TGF-β增高的表达水平可被醛糖还原酶抑制剂所抑制并且呈剂量依赖(P<0.05)。结论糖尿病腰椎管狭窄患者黄韧带中山梨醇水平显著增高,进而促进炎性反应因子及纤维化相关因子TGF-β表达增加,使得黄韧带炎性增生。

Objective To investigate the related mechanism of ligamentum flavum（ LF） hypertrophy in diabetic patients with lumbar spinal canal stenosis（ LSCS）. Methods Twenty-four diabetes mellitus patients [DM（ ＋） ]and twenty normoglycemic patients [DM（-） ]with LSCS were enrolled in this study. Sorbitol in LF was analyzed using D-Sorbitol / Xylitol test kit. The thickness of LF was measured by CT. The structure of LF was observed after HE and Masson＇s trichrome staining. The cell cycle and proliferation of fibroblastic cell NIH3T3 line cultured in high glucose were analyzed. Sorbitol of NIH3T3 was detected under different backgrounds in vitro,normal glucose,high glucose and high glucose burdened with aldose reductase inhibitor（ ARI）,Epalrestat. The expression of inflammatory factors was detected by q PCR and Western blot under above different backgrounds. Results LF of diabetic patients exhibited significantly higher level of sorbitol and pro-inflammatory cytokines,TGF-β and of CD68-positive staining than that of the normoglycemic subjects（ P 0. 01）. The diabetic LF was significantly thicker than that of the controls,and showed evidence of degeneration. The high glucose-cultured fibroblasts exhibited significantly higher levels ofsorbitol,pro-inflammatory factors,and TGF-β compared to the low glucose-cultured cells,and these levels were dose-dependently reduced by treatment with the aldose reductase inhibitor（ P 0. 05）. Conclusions Sorbitol level of the LF is significantly increased in the DM patients with LSCS. Increased sorbitol recruites inflammatory factors and fibrogenic-related factor TGF-β in LF of DM patients with LSCS which may contributes to the LF hypertrophy.