AIM: To investigate the regulation of activin receptor-interacting protein 2 (ARIP2) expression and its possible relationships with collagen type Ⅳ (collagen Ⅳ) in mouse hepatoma cell line Hepal-6 cells. METHOD...AIM: To investigate the regulation of activin receptor-interacting protein 2 (ARIP2) expression and its possible relationships with collagen type Ⅳ (collagen Ⅳ) in mouse hepatoma cell line Hepal-6 cells. METHODS: The ARIP2 mRNA expression kinetics in Hepal-6 cells was detected by RT-PCR, and its regulation factors were analyzed by treatment with signal transduction activators such as phorbol 12-myristate 13-acetate (PMA), forskolin and A23187. After pcDNA3- ARIP2 was transfected into Hepal-6 cells, the effects of ARIP2 overexpression on activin type Ⅱ receptor (ActRⅡ) and collagen Ⅳ expression were evaluated. RESULTS: The expression levels of ARIP2 mRNA in Hapel-6 cells were elevated in time-dependent manner 12 h after treatment with activin A and endotoxin LPS, but not changed evidently in the early stage of stimulation (2 or 4 h). The ARIP2 mRNA expression was increased after stimulated with signal transduction activators such as PMA and forskolin in Hepal-6 cells, whereas decreased after treatment with A23187 (25.3% ± 5.7% vs 48.1% ± 3.6%, P 〈 0.01). ARIP2 overexpression could remarkably suppress the expression of ActRⅡA mRNA in dose-dependent manner, but has no effect on ActRⅡB in Hepal-6 cells induced by activin A. Furthermore, we have found that overexpression of ARIP2 could inhibit collagen Ⅳ mRNA and protein expressions induced by activin A in Hapel-6 cells. CONCLUSION: These findings suggest that ARIP2 expression can be influenced by various factors. ARIP2 may participate in the negative feedback regulation of signal transduction in the late stage by affecting the expression of ActRIIA and play an important role in regulation of development of liver fibrosis induced by activin.展开更多
采用DEAE-FF离子交换和Sephdax G-15葡聚糖凝胶层析方法,从玉米肽中分离、纯化,获得具有抑制HepG2细胞增殖活性的短肽,采用高效液相色谱串联质谱电喷雾法(liquid chromatography-electrospray ionization tandem mass spectrometry,LC-E...采用DEAE-FF离子交换和Sephdax G-15葡聚糖凝胶层析方法,从玉米肽中分离、纯化,获得具有抑制HepG2细胞增殖活性的短肽,采用高效液相色谱串联质谱电喷雾法(liquid chromatography-electrospray ionization tandem mass spectrometry,LC-ESI-MS/MS)对其活性片段进行结构鉴定。结果表明:其短肽的一级结构为LPPYLP[命名为玉米六肽(corn peptides-6,CPs-6)],通过四唑盐[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT]比色法试验发现,CPs-6具有抑制HepG2细胞增殖的效果,且对L-02细胞的毒性低。展开更多
基金Supported by the National Natural Science Foundation of China, No. 30170478 and 30571688Science Projects of Jilin Province of China, No. 20060928-01
文摘AIM: To investigate the regulation of activin receptor-interacting protein 2 (ARIP2) expression and its possible relationships with collagen type Ⅳ (collagen Ⅳ) in mouse hepatoma cell line Hepal-6 cells. METHODS: The ARIP2 mRNA expression kinetics in Hepal-6 cells was detected by RT-PCR, and its regulation factors were analyzed by treatment with signal transduction activators such as phorbol 12-myristate 13-acetate (PMA), forskolin and A23187. After pcDNA3- ARIP2 was transfected into Hepal-6 cells, the effects of ARIP2 overexpression on activin type Ⅱ receptor (ActRⅡ) and collagen Ⅳ expression were evaluated. RESULTS: The expression levels of ARIP2 mRNA in Hapel-6 cells were elevated in time-dependent manner 12 h after treatment with activin A and endotoxin LPS, but not changed evidently in the early stage of stimulation (2 or 4 h). The ARIP2 mRNA expression was increased after stimulated with signal transduction activators such as PMA and forskolin in Hepal-6 cells, whereas decreased after treatment with A23187 (25.3% ± 5.7% vs 48.1% ± 3.6%, P 〈 0.01). ARIP2 overexpression could remarkably suppress the expression of ActRⅡA mRNA in dose-dependent manner, but has no effect on ActRⅡB in Hepal-6 cells induced by activin A. Furthermore, we have found that overexpression of ARIP2 could inhibit collagen Ⅳ mRNA and protein expressions induced by activin A in Hapel-6 cells. CONCLUSION: These findings suggest that ARIP2 expression can be influenced by various factors. ARIP2 may participate in the negative feedback regulation of signal transduction in the late stage by affecting the expression of ActRIIA and play an important role in regulation of development of liver fibrosis induced by activin.
文摘采用DEAE-FF离子交换和Sephdax G-15葡聚糖凝胶层析方法,从玉米肽中分离、纯化,获得具有抑制HepG2细胞增殖活性的短肽,采用高效液相色谱串联质谱电喷雾法(liquid chromatography-electrospray ionization tandem mass spectrometry,LC-ESI-MS/MS)对其活性片段进行结构鉴定。结果表明:其短肽的一级结构为LPPYLP[命名为玉米六肽(corn peptides-6,CPs-6)],通过四唑盐[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT]比色法试验发现,CPs-6具有抑制HepG2细胞增殖的效果,且对L-02细胞的毒性低。