Ulcerative Colitis: Gan H

Chen Y, Gan H, Ouyang Q, Xu D, Pan Y, A Z. · Department of Life Sciences, Dali College, Dali 671000, China. · Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. · Pubmed #15553846 No free full text.

Abstract: The purpose of this study is to assess the effects of anti-inflammatory on activation of nuclear factor-kappaB and mRNA and protein expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in intestinal mucosal biopsy specimens from patients with ulcerative colitis (UC). A total of 27 cases with UC were investigated. 15 cases received sulfasalazine (SASP) treatment or SASP and glucocorticoid treatment, 12 cases did not receive any medication related with UC. Normal mucosa from 9 colon cancer cases served as control. Ten pieces of intestinal mucosal biopsy specimens were obtained from each patient. The mRNA expression of ICAM-1 and VCAM-1 were determined by reversal transcription-polymerase chain reaction (RT-PCR). The protein levels of ICAM-1 and VCAM-1 were measured by enzyme linked immunosorbent assay (ELISA). NF-kappaB DNA binding activity was evaluated by electrophoretic mobility shift assay (EMSA). The results showed that NF-kappaB DNA binding activity, mRNA and protein expression of ICAM-1 and VCAM-1 were increased significantly in patients with UC, compared with normal control (P<0.05). Glucocorticoids and SASP markedly inhibited NF-kappaB activation and significantly decreased mRNA and protein expression of ICAM-1 and VCAM-1 (P<0.05). Adhesion molecules (ICAM-1 and VCAM-1) gene activation had significant positive correlation with the NF-kappaB DNA binding activity (r=0.8652 P<0.05, r=0.7902, P<0.05, respectively). We concluded that NF-kappaB is a major and essential factor in regulating the expression of adhesion molecules, it plays an important role in the pathogenesis of UC. SASP and glucocorticoids ameliorate UC via inhibition of NF-kappaB activation and reduction of adhesion molecules expression.

Gan H, Ouyang Q, Chen Y, Liang F. · Department of Gastroenterology, West China Hospital, Sichuan University, Chengdu 610041. · Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. · Pubmed #12856595 No free full text.

Abstract: To investigate if nuclear factor-kappa B (NF-kappa B) p65 antisense oligonucleotides might affect the expression of NF-kappa B p65 and cytokines in lamina propria mononuclear cells(LPMC) from patients with ulcerative colitis (UC). LPMC were isolated from intestinal mucosal biopsy specimens from 3 patients with UC, and cultured with or without NF-kappa B p65 antisense oligonucleotides (5'-GGAACAGTTCGTCCTATGG-3'), missense oligonucleotides (5'-GGAACAGTTCGTCTATGG-3') and dexamethasone. NF-kappa B p65 expression was determined by western blot analysis. The expression of cytokine mRNA was studied by reversal transcription-polymerase chain reaction (RT-PCR). The cytokine levels were measured by enzyme linked immunosorbent assay. The results showed that NF-kappa B p65 antisense oligonucleotides resulted in down-regulation of NF-kappa B p65 expression, blocked the expression of IL-1 beta mRNA and IL-8 mRNA, and strikingly reduced the production of IL-1 beta and IL-8, and these effects were greater than those of dexamethasone in cultured LPMC from patients with UC(P < 0.05). Therefore, the application of NF-kappa B p65 antisense oligonucleotides may serve as a novel molecular approach for the treatment of patients with UC.

Gan H, Ouyang Q, Jia D, Xia Q. · Department of Gastroenterology, West China Hospital, Sichuan University, Chengdu 610041, China. · Zhonghua Nei Ke Za Zhi. · Pubmed #12133438 No free full text.

Abstract: OBJECTIVE: To investigate the activation of nuclear factor-kappaB (NF-kappaB) and its relationship with expression of cytokine mRNA in intestinal mucosal biopsy specimens from patients with ulcerative colitis (UC). METHODS: 31 cases with UC were included in the study. 17 cases received sulfasalazine (SASP) or SASP and glucocorticoid treatment. 14 cases did not receive any medication related with UC. Normal mucosa from 11 colon cancer cases served as control. Ten pieces of intestinal mucosal biopsy specimens were obtained from each patient. NF-kappaB DNA binding activity was evaluated with electrophoretic mobility shift assay (EMSA). Expression of cytokine mRNA were studied with reversal tanscription-polymerase chain reaction (RT-PCR). RESULTS: (1) The expression of IL-1beta mRNA and IL-8 mRNA was increased significantly in patients with UC, as compared with that in the control specimens (P < 0.05) and had a significant positive correlation with NF-kappaB DNA binding activity (r = 0.8363, P < 0.05; r = 0.6024, P < 0.05, respectively). (2) Glucocorticoids and SASP strongly inhibited NF-kappaB activation and signficantly decreased the expression of IL-1beta mRNA and IL-8 mRNA. CONCLUSIONS: NF-kappaB is a major and essential factor in regulating the expression of cytokine and plays a fundamental role in the pathogenesis of UC. SASP and glucocorticoids decrease cytokine expression via inhibition of NF-kappaB activation.

Gan H, Ouyang Q, Chen Y, Xia Q. · Department of Gastroenterology, The West China Hospital Sichuang University, Chengdu 610041, China. · Zhonghua Yi Xue Za Zhi. · Pubmed #11953203 No free full text.

Abstract: OBJECTIVE: To investigate the activation and expression of nuclear factor-kappaB (NF-kappaB) and effects of anti-inflammatory treatment on NF-kappaB in the intestinal mucosa of patients with ulcerative colitis (UC). METHODS: Ten pieces of colon mucosal biopsy specimens were obtained from 31 cases with UC, 17 of which received sulphasalazine (SASP) or SASP plus glucocorticoid and 14 of which received no medication. Samples of normal mucosa around the lesion taken from 11 patients with colon cancer were used as controls. NF-kappaB DNA binding activity was evaluated by electrophoretic mobility shift assay. NF-kappaB p65 expression was determined by Western blot analysis and immunohistochemical staining with a NF-kappaB p65 antibody. The type of cells containing activated NF-kappaBp65 was identified by double immunofluorescence confocal laser scanning microscopy. RESULTS: The expression of NF-kappaB p65 and NF-kappaB DNA binding activity were significantly higher in patients with UC than in the control (P < 0.05), and were correlated with the degree of inflammation. The NF-kappaB expression was significantly stronger in the nuclei than in the cytoplasm in patients with UC without pharmacotherapy. The NF-kappaB expression in nuclei was significantly stronger in the group without pharmacotherapy than in the group with pharmacotherapy (P < 0.05). Only a few NF-kappaB p65 positive cells were seen in the controls. NF-kappaBp65 expression was found in all major subsets of mononuclear cells, including macrophages, B lymphocytes, T lymphocytes, and cryptal epithelial cells. CONCLUSION: The increased activation of NF-kappaB and increased expression of NF-kappaB may be involved in the pathogenesis of UC. Glucocorticoids and SASP strongly inhibited NF-kappaB activation and expression. The inhibition of NF-kappaB activation may be a central part of the anti-inflammatory action of glucocorticoids and SASP, which might represent an important pharmacological mechanism in treatment of patients with UC. NF-kappaB will be an important target for cytokine-based therapy of UC.