Study for implication of apoptotic mechanism in massive hepatic cell death and its control

细胞凋亡机制在肝细胞大量死亡及其调控中的意义研究

• 批准号: 14570500
• 负责人: TAKIKAWA Yasuhiro
• 金额: $ 0.9万
• 依托单位: Iwate Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570500/
• 关键词: Hepatocyte apoptosis; Phosphatidyl serine; Blood coagulation; Massive hepatocyte death; Pros tagland in; Bcl-xL; 劇症肝炎; 肝細胞; アポトーシス

1. Acceleration of blood coagulation by apoptotic hepatocellular carcinoma HepG2 cells1) The apoptosis of Hep G2 cells was induced by staurosporin (STS).2) Phosphatidyl serine (PS) expression was detected by confocal microscopy with FITC-annexin V, a specific inhibitor of PS activity.3) The STS treatment of HepG2 cells significantly accelerated thrombin generation, which was determined using a thrombin-specific chromogenic substrate after exposing the cells to activated factor X, CaCl_2 and prothrombin. 4) The increase in thrombin generation by STS treatment was abolished following pretreatment with annexin V.These data indicate that apoptotic hepatocytes accelerate blood coagulation through the expression of a PS-dependent procoagulant surface.2. Prostaglandin receptor EP4 agonist (GPEP4A) suppression of Fas-induced apoptosis in HepG2 cells through direct Bcl-xL induction.1) GPEP4A suppressed Fas-induced apoptosis of HepG2 cells, whereas indomethacin accelerated it.2) PGE2RA significantly increased the expression levels of the Bcl-xL protein and mRNA; indomethacin decreased the expression levels, of Mdl proteins.These data indicate that direct Bcl-xL induction plays an important role in the hepatocyte protective effect induced by PGE2R-A.

1. 肝癌细胞凋亡对HepG2细胞的加速凝血作用1)staurosporin （STS）诱导HepG2细胞凋亡。2)用特异性磷脂酰丝氨酸活性抑制剂FITC-annexin V共聚焦显微镜检测磷脂酰丝氨酸（PS）的表达。3)经STS处理的HepG2细胞在活化因子X、CaCl_2和凝血酶原的作用下，凝血酶的生成明显加快，采用凝血酶特异性显色底物测定。4)经STS处理的凝血酶生成的增加在膜联蛋白v预处理后被消除。这些数据表明，凋亡的肝细胞通过表达ps依赖性促凝表面来加速凝血。前列腺素受体EP4激动剂（GPEP4A）通过Bcl-xL直接诱导抑制fas诱导的HepG2细胞凋亡。1) GPEP4A抑制fas诱导的HepG2细胞凋亡，而吲哚美辛则加速其凋亡。2) PGE2RA显著提高Bcl-xL蛋白和mRNA的表达水平；吲哚美辛降低了Mdl蛋白的表达水平。这些数据表明，直接诱导Bcl-xL在PGE2R-A诱导的肝细胞保护作用中起重要作用。

项目成果

Lin, SD, Endo, R, Takikawa, Y, et al.: "Plasma and urine levels of urinary tripsin inhibitor in patients with acute and fulminant hepatitis"J Gastroenterol Hepatol. 17. 140-147 (2002)

Lin, SD, Endo, R, Takikawa, Y 等人：“急性和暴发性肝炎患者尿胰蛋白酶抑制剂的血浆和尿液水平”J Gastroenterol Hepatol。

滝川康裕, 鈴木一幸: "劇症肝炎・LOHFの現況. 内科的治療"日本消化器病学会誌. 99・7. 905-912 (2002)

泷川泰宏、铃木一之：“暴发性肝炎/LOHF的现状。内部治疗”日本胃肠病学会杂志99・7（2002年）。

Takikawa, Y, Suzuki, K: "Is AFP a new reliable marker of liver regeneration in acute hepatic failure?"J Gastroenterology. 37. 681-682 (2002)

Takikawa, Y, Suzuki, K：“AFP 是急性肝衰竭中肝再生的新可靠标志物吗？”J Gastroenterology。

A.Ushio, Y Takikawa, et al.: "Induction of Bcl-xL is a possible mechanism of antiapoptotic effect by prostaglandin E2 EP4-receptor agonist in human hepatocellular carcinoma HepG2 cells"Hepatology Research. (印刷中).

A.Ushio、Y Takikawa 等人：“Bcl-xL 的诱导是前列腺素 E2 EP4 受体激动剂在人肝细胞癌 HepG2 细胞中产生抗凋亡作用的可能机制”肝病学研究（正在出版）。

Y.Miyamoto, Y Takikawa, et al.: "Apoptotic hepatocellular carcinoma HepG2 cells accelerate blood coagulation"Hepatology Research. (in press). (2004)

Y.Miyamoto，Y Takikawa，等：“凋亡性肝细胞癌HepG2细胞加速血液凝固”肝病学研究。