Reg Ⅳ在溃疡性结肠炎中的表达及其对疾病活动度的预测价值

背景再生基因Ⅳ(Regenerating gene Ⅳ,RegⅣ)是钙依赖的植物血凝素超家族成员之一,主要位于胃肠道。目前缺乏RegⅣ与溃疡性结肠炎关系的研究报告。目的探讨RegⅣ与C反应蛋白、溃疡性结肠炎(ulcerative colitis,UC)活动度之间的关系。方法收集32例UC及10例健康查体者的肠黏膜及血清标本,通过免疫组化检测RegⅣ蛋白的表达水平。结果免疫组化染色提示Reg Ⅳ在UC患者肠黏膜中的表达水平较正常人明显升高(P0.05)。The colitis activity index(CAI)与C反应蛋白呈中度正相关(r=0.529,P0.01)。结论RegⅣ可作为预测UC活动程度的分子标志。     

Prospects and Challenges for Islet Regeneration as a Treatment for Diabetes: A Review of Islet Neogenesis Associated Protein

Diabetes mellitus results from inadequate insulin action, which can be viewed as a consequence of the limited ability to restore β cells after they are lost as the result of metabolic exhaustion, autoimmune destruction, or surgical insult. Arguably, a uniformly effective therapeutic pathway to address all forms of diabetes would be to reverse the restrictions on β-cell and islet regeneration. The development from progenitor cells of islets with normal endocrine function does occur in adult humans; it is referred to as islet neogenesis. The induction of islet neogenesis is an important, if not essential, therapeutic approach for curing type 1 diabetes mellitus (T1DM) and could be valuable in the treatment of type 2 diabetes mellitus (T2DM) as well. Islet neogenesis associated protein (INGAP) is the first therapeutic candidate to be identified as the result of a purposeful search for an endogenous molecule with islet neogenic activity. It was found that partial obstruction of the pancreatic duct in hamsters induced islet neogenesis; under this condition, a neogenesis-promoting activity was identified and partially purified from a soluble tissue fraction. A 168-kDa protein product of the cloned gene was found to be responsible for the neogenesis activity. This molecule named INGAP contains an active core sequence of amino acids called INGAP peptide. Results from in vitro, animal, and human studies suggest that INGAP and INGAP peptide are neogenic in at least several vertebrate species, including humans. INGAP has since been found to be a member of the family of Reg proteins, which are found across and in multiple versions within species and are closely associated with embryonic and regenerative processes. Clinical results suggest that INGAP peptide can be a suitable neogenesis therapy, but optimization of the therapy and more data are required to fully access this potential. Understanding of the signaling pathways of INGAP and other related Reg proteins is a promising means of advancing therapeutic development for people with T1DM and T2DM. The quest for the fundamental restorative approach to lost insulin secretion is an enticing target for drug development.     

microRNA-24靶向RegⅣ抑制胃癌细胞生长与转移

目的:探讨microRNA-24(miR-24)在胃癌中的作用及其作用靶基因。方法 :采用qRT-PCR的方法检测miR-24在9株胃癌细胞和永生化胃黏膜上皮细胞GES-1中的表达,同时检测63例病人的胃癌组织及相应的癌旁正常组织中miR-24的表达,分析miR-24与胃癌临床病理特征之间的关系。应用miR-24模拟体转染胃癌细胞株,使其过表达miR-24,细胞增殖试验(CCK-8法)、流式细胞技术(Annexin-Ⅴ/PI标记)、Transwell试验检测上调胃癌细胞株miR-24表达后细胞增殖、凋亡、迁移和周期的变化;并用ELISA技术和双荧光素酶基因报告载体检测miR-24是否调控RegⅣ的表达。结果:54.0%(34/63)胃癌病人的miR-24在肿瘤组织中表达,与相应的癌旁正常组织相比下降,且miR-24在肿瘤组织中的表达与胃癌的组织分化程度相关(r=0.292,P=0.021),与其他的临床病例特征如年龄、性别、浸润深度、淋巴结转移和TNM分期没有相关性。miR-24抑制胃癌细胞的生长与转移,且促进胃癌细胞的凋亡。miR-24可降低RegⅣ的蛋白表达水平。结论:miR-24抑制胃癌的发生、发展,miR-24可能通过调控RegⅣ的表达来发挥其生物学效应。RegⅣ与miR-24均在胃癌的发生、发展中发挥了重要作用。     

Effect of a gastrin/cholecystokinin B receptor antagonist, S-0509, on the omeprazole-induced proliferation of gastric mucosa in rats

Hypergastrinemia is known to cause hyperplasia of the gastric mucosa, especially in gastric enterochromaffinlike (ECL) cells. In some clinical conditions causing hypergastrinemia, such as long-term gastric-acid inhibition and gastric-mucosa atrophy, hyperplastic ECL cells may develop into gastric carcinoid tumors. A newly developed gastrin-receptor antagonist, S-0509, has been reported to block gastrin-induced stimulation of gastric-acid secretion. We therefore investigated whether S-0509 inhibits the omeprazole- and gastrin-stimulated hyperproliferation of gastric mucosa, especially of ECL cells. Daily administration of omeprazole and gastrin in male Sprague-Dawley rats induced marked hypergastrinemia and increased proliferation of gastric-mucosa cells. The numbers of ECL cells and of ECL cells producing messenger RNA for regenerating gene, a potent growth factor for gastric-mucosa cells, were also augmented by long-term administration of omeprazole and gastrin. Coadministration of S-0509 with omeprazole or gastrin almost completely inhibited the omeprazole- and gastrin-induced changes in gastric mucosa, including mucosal thickening and ECL hyperplasia. S-0509 did not induce gastric-mucosa atrophy, even when administered for as long as 4 weeks. In summary, we have found that a newly developed gastrin receptor antagonist, S-0509, inhibits omeprazole- and gastrin-induced mucosal hyperplasia, especially ECL-cell hyperplasia, in rats.     

Reg Ⅳ, a differentially expressed gene in colorectal adenoma

Objective To discover and identify differentially expressed genes associated with colorectal adenomaforma tion and the role of ReglV in colorectal adenoma differentiation.Methods A subtracted cDNA library was constructed with cDNAs that were isolated from either the normal mucosa or adenoma tissue of a single patient. Suppressive subtractive hybridization (SSH)combined with virtual northern blotting was used to characterize differentially expressed genes and contigs were assembled by electronic cloning (in silico cloning ) with the EST database. Semi-quantitative RT-PCR was performed in 9 colorectal adenomas.Results The amino acid sequence was determined with open reading frame (ORF) prediction software and was found to be 100% homologous to the protein product of Reg Ⅳ (a novel gene isolated from a large inflammatory bowel disease library). RegⅣ was found to be highly expressed in all of the adenoma samples (9/9) compared with the normal mucosa samples, while 5/6 casess howed RegⅣ to be more strongly expressed in adenocarcinoma.Conclusion RegⅣ may play an important role in the initiation of colorectal adenoma differentiation,and its detection mav be useful in the earlv diaonosis of colorectal adenoma formation.     

Pancreatic regenerating protein (regⅠ) and regⅠreceptor mRNA are upregulated in rat pancreas after induction of acute pancreatitis

AIM: Pancreatic regenerating protein (regⅠ) stimulates pancreatic regeneration after pancreatectomy and is mitogenic to ductal andβ-cells. This suggests that regⅠand its receptor may play a role in recovery after pancreatic injury. We hypothesized that regⅠand its receptor are induced in acute pancreatitis. METHODS: Acute pancreatitis was induced in male Wistar rats by retrograde injection of 3% sodium taurocholate into the pancreatic duct. Pancreata and serum were collected 12, 24, and 36 hours after injection and from normal controls (4 rats/group). RegⅠreceptor mRNA, serum regⅠprotein, and tissue regⅠprotein levels were determined by Northern analysis, enzyme-linked immunosorbent assay (ELISA), and Western analysis, respectively. Immunohistochemistry was used to localize changes in regⅠand its receptor. RESULTS: Serum amylase levels and histology confirmed necrotizing pancreatitis in taurocholate treated rats. There was no statistically significant change in serum regⅠconcentrations from controls. However, Western blot demonstrated increased tissue levels of regⅠat 24 and 36 h. This increase was localized primarily to the acinar cells and the ductal cells by immunohistochemistry. Northern blot demonstrated a significant increase in regⅠreceptor mRNA expression with pancreatitis. Immunohistochemistry localized this increase to the ductal cells, islets, and acinar cells. CONCLUSION: Acute pancreatitis results in increased tissue regⅠprotein levels localized to the acinar and ductal cells, and a parallel threefold induction of regⅠreceptor in the ductal cells, islets, and acinar cells. These changes suggest that induction of reg I and its receptor may be important for recovery from acute pancreatitis.