von Boyen G B T, Steinkamp M, Reinshagen M, Schäfer K-H, Adler G, Kirsch J
Department of Medicine I (Gastroenterology), University of Ulm, Albert-Einstein-Allee 11, 89069 Ulm, Germany.
Gut. 2004 Feb;53(2):222-8. doi: 10.1136/gut.2003.012625.
Enteric glia protect the integrity of the gut, as loss of enteric glial fibrillary acidic protein (GFAP) positive (+) glia leads to a haemorrhagic jejunoileitis. Crohn's disease (CD) and necrotising enterocolitis (NEC) show pathological changes in enteric glia. Therefore, factors controlling GFAP+ enteric glia are of great interest. The aim of the present study was to characterise enteric glia and determine the effect of interleukin 1beta (IL-1beta), interleukin 4 (IL-4), tumour necrosis factor alpha (TNF-alpha), and lipopolysaccharides (LPS) on cultured enteric glia.
Dissected rat colon and cultured enteric glia cells were double labelled with anti-GFAP and anti-S-100 antibodies. For regulatory studies, enteric glia cells were treated with cytokines and LPS. Proliferation was assayed using bromodeoxyuridine (BrdU) and mitosis of enteric glia was blocked by demecolcine.
We were able to distinguish GFAP negative (-) from GFAP+ glia subtypes in situ and in primary cultures. Incubation of cells with IL-1beta, TNF-alpha, and LPS led to a significant increase in GFAP+ enteric glia while IL-4 had no effect on GFAP expression. After incubation with IL-1beta, total intracellular GFAP of enteric glia cells was increased. Upregulation of GFAP+ enteric glia could also be observed after stimulation with IL-1beta on blocking mitosis. BrdU uptake in stimulated enteric glia showed no increased proliferation rate.
Two different types of enteric glia based on GFAP expression exist in the gut. Proinflammatory cytokines and LPS cause a dramatic increase in GFAP+ enteric glia. This suggests that cytokines play an important role in controlling GFAP+ enteric glia which might in turn be involved in modulating the integrity of the bowel during inflammation.
肠神经胶质细胞可保护肠道的完整性，因为肠神经胶质纤维酸性蛋白（GFAP）阳性（+）的神经胶质细胞缺失会导致出血性空肠回肠炎。克罗恩病（CD）和坏死性小肠结肠炎（NEC）均表现出肠神经胶质细胞的病理变化。因此，控制GFAP+肠神经胶质细胞的因素备受关注。本研究的目的是对肠神经胶质细胞进行表征，并确定白细胞介素1β（IL-1β）、白细胞介素4（IL-4）、肿瘤坏死因子α（TNF-α）和脂多糖（LPS）对培养的肠神经胶质细胞的影响。
用抗GFAP和抗S-100抗体对解剖后的大鼠结肠和培养的肠神经胶质细胞进行双重标记。为进行调控研究，用细胞因子和LPS处理肠神经胶质细胞。使用溴脱氧尿苷（BrdU）检测增殖情况，并用秋水仙胺阻断肠神经胶质细胞的有丝分裂。
我们能够在原位和原代培养中区分GFAP阴性（-）和GFAP+神经胶质细胞亚型。用IL-1β、TNF-α和LPS孵育细胞导致GFAP+肠神经胶质细胞显著增加，而IL-4对GFAP表达无影响。用IL-1β孵育后，肠神经胶质细胞的总细胞内GFAP增加。在阻断有丝分裂的情况下，用IL-1β刺激后也可观察到GFAP+肠神经胶质细胞的上调。刺激后的肠神经胶质细胞中BrdU摄取显示增殖率未增加。
肠道中存在基于GFAP表达的两种不同类型的肠神经胶质细胞。促炎细胞因子和LPS导致GFAP+肠神经胶质细胞显著增加。这表明细胞因子在控制GFAP+肠神经胶质细胞中起重要作用，而这可能反过来参与调节炎症期间肠道的完整性。
