Identification of HGF indncing factor in serum from patients with Liver diseases

肝病患者血清中HGF诱导因子的鉴定

• 批准号: 05454247
• 负责人: TSUBOUCHI Hirohito
• 金额: $ 4.29万
• 依托单位: Miyazaki Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05454247/
• 关键词: HGF; HGF inducing factor; HGF mRNA; Liver disease; HGFmRNA; HGF産生誘導因子; MRC-5細胞; サイトカイン

MRC-5 cells were plated on 24-well plastic dish in 0.5 ml of DMEM supplemented with 10% FCS and antibiotics, and cultured at 37 ﾟC in a humidified atmosphere of 5% CO_2 in the presence or absence of patient's serum. Serum samples were obtained from normal subjects, patients with fulminant hepatic failure (FH), acute hepatitis (AH), chronic hepatitis (CH), liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Serum samples were added to the culture medium at a final concentration of 5% and the cells were cultured for 24 hours after the addition of samples. Hepatocyte growth factor (HGF) in the culture supernatant was determined using an enzyme-linked immunosorbent assay (ELISA) and HGF production was expressed as HGF amount per cellular protein (ng/mg protein).Sera from patients with AH and CH increased HGF production in MRC-5 cells with dose- and time-dependent manner. The relationship between HGF production by patients' serum and HGF levels in serum of AH and CH patients was positively significant. Northern blot analysis revealed an increase in HGF mRNA levels of MRC-5 cells cultured with serum from AH patients. HGF-inducing activity in the patients' serum was disappeared by heat treatment (56ﾟC,30min) and treatment with trypsin or chymotrypsin. We also examined the effects of interleukin-1 (IL-1), interleukin-2 (IL-2), tumor necrosis factor-alpha (TNF-alpha), phorbol ester (PMA), dexamethasone and heparin on HGF production in MRC-5 cells cultured with serum having a high HGF- inducing activity. IL-1, TNF-alpha and PMA had an additive effect on HGF production stimulated by patient's serum in MRC-5 cells, however, dexamethasone suppressed the HGF production.In summarize, HGF-inducing activity was present in serum of patients with various liver diseases and this activity may be a physical HGF inducer in humans.

将MRC细胞铺在24孔塑料盘中，在0.5 mL的DMEM中，在有10％FC的DMEM中，在存在或不存在患者血清样品的情况下以37个球体培养5％CO_2。肝衰竭（FH），急性肝炎（AH），肝硬化（LC）ND ND肝细胞癌（HCC）。使用酶线的IMUNOSORBENT分析（ELISA）和HGF产生以HGF量表现为HGF量表现蛋白（NG/mg蛋白）。依赖性的方式。检查了所检查的所检查的TOKIN-1（IL-1）ORBOR酯（PMA），地塞米松和肝素在MRC-5培养的MRC-5中产生的HGF产生，其MRC-5中患者血清模拟了HGH HGF-IL-1然而，细胞抑制了HGF的产生。总而言之，HGF-二率存在于偏爱的肝脏疾病的血清中，而thysivity则可能是人类的物理HGF ucer。

项目成果

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S.Kawakami, H.Tsubouchi, S.Nakagawa, et al.: "Expression of Hepatocyte Growth Factor in Normal and Carbone Tetrachloride-treated Monkeys." Hepatology. 20. 1255-1260 (1994)

S.Kawakami、H.Tsubouchi、S.Nakakawa 等人：“正常和四氯化碳处理的猴子中肝细胞生长因子的表达”。