Hepatitis B Virus X Protein Upregulates Tumor Necrosis Factor-? Expression of Rat Mesangial Cell Line Via Extracellular Regulated Protein Kinases Pathway
- VernacularTitle:乙型肝炎病毒X蛋白通过激活细胞外蛋白调节激酶转导机制上调大鼠系膜细胞肿瘤坏死因子-?的表达
- Author:
hong-zhu, LU
;
jian-hua, ZHOU
- Publication Type:Journal Article
- Keywords:
hepatitis B virus;
X gene;
mesangial cell line;
extracellular regulated protein kinases;
tumor necrosis factor-?
- From:Journal of Applied Clinical Pediatrics
2003;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the extracellular regulated protein kinases (ERKs) pathway of hepatitis B virus(HBV) X protein(HBx) on glomerular mesangial cell(GMC) proliferation of rat and tumor necrosis factor(TNF)-? expression.Methods The HBV X gene was amplified by polymerase chain reaction(PCR),inserted into the eukaryotic expression vector PCI-neo and confirmed by restrict endonuclease digestion and sequence analysis.PCI-neo contained HBV X gene (PCI-neo-X) was transfected into cultured GMC via liposome.GMC proliferation,TNF-? and its mRNA expression were investigated in the condition of with or without U0126 in the culture media.HBx,ERK1/2 and phosphorynated-ERK1/2 (p-ERK1/2) expression in GMC were assessed by Western blot.TNF-? mRNA expression was assessed by semi-quantitative reverse transcriptase-PCR (RT-PCR).TNF-? level in supernatants was measured by enzymelinked immunosorbent assay(ELISA).GMC proliferation was assessed by methyl thiazolyl tetrazolium(MTT).Results HBx expression was found in transfected GMC,and became prominent at 36 and 48 hours after transfection whether with or without U0126 in culture media.TNF-? mRNA expression was significantly decreased in U0126 group compared with U0126-free group.TNF-? levels in supernatants in PCI-neo-X transfection without U0126 group were (189.0?18.1) ng/L at 36 hours and (172.3?24.3) ng/L at 48 hours after transfection,respectively.In contrast,TNF-? levels in supernatants with U0126 were (65.6?11.6) ng/L and (84.0?24.6) ng/L,respectively.The TNF-? le-vels in the latter groups were significantly lower than the formers (Pa
