Expression of nuclear factor kappa B and inducible nitric oxide synthase in the respiratory syncytial virus infection on human lung epithelial cells
- VernacularTitle:人肺上皮细胞感染呼吸道合胞病毒后核内活性因子?B与诱导型一氧化氮合酶的表达
- Author:
Wei LIU
;
Yun YUN
;
Shenghai HUANG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2007;0(15):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: After human lung epithelial cells (A549) infected with respiratory syncytial virus (RSV), inducer cells over-express active transmitter. The study investigated the expression changes in nuclear factor ?B (NF-?B), inducible nitric oxide synthase (iNOS) mRNA and protein, nitric oxide (NO) and malonaldehyde (MDA) on human lung epithelial cells (A549) infected with RSV. METHODS: Experiments were performed at the Laboratory of Etiology and Immunology, Anhui Medical University between December 2006 and May 2007. ① Gyopreserved virus broliferated in Hep-2 cells after recovery. A549 cells infected with RSV in vitro were used to collect cells and cellular culture supernatant at hours 4, 8, 16 and 24. Cells non-infected with RSV were served as controls. ② Reverse transcription-polymerase chain reaction (RT-PCR) was used to evaluate the expression of iNOS mRNA. The expression of iNOS protein and NF-?B was detected by Western-Blot. The concentrations of NO and MDA were measured by nitrate reductase method and thiobarbituric acid method. RESULTS: ①The basal expression of NF-?B was increased with the prolongation of RSV infection to A549 cells. ②RSV infection to A549 increased the amounts of mRNA and protein of iNOS in a time-dependent manner. The expression of iNOS mRNA was about 12 times as many as basal expression. RSV infection promoted the expression of iNOS protein, which was higher than the control group. The activities of NF-?B significantly were positively correlated with the mRNA and protein expression of iNOS. ③RSV could improve A549 cells secreting NO and MDA, but the NO levels rose slowly during the 24 hours of infection. CONCLUSION: The inflammatory response of RSV can increase the activity of NF-?B and it is positive correlated with the increase in iNOS mRNA and protein levels.
