ISSN: 0973-7510

E-ISSN: 2581-690X

Jian da Zhou1, Pashupati Babu Pokharel2, Feng Hu1, Wu Xiong1, Ke Cao3, Jin-yan Liu1, Bi-mei Jiang4, Yong Chen5, Rui Liu1 and Can-juan Xiong6
1Department of Plastic and Reconstructive Surgery, Third Xiangya Hospital.
2Department of Plastic and reconstructive Surgery, Xiang ya Hospital.
3Department of Oncology, Third Xiangya Hospital.
4Xiangya School, Central South University, Changsha, China.
5Family Planning Institute of Hunan Province, Changsha, China.
6Third Xiangya Hospital, Central South University, Changsha ,China.
J Pure Appl Microbiol. 2013;7(Spl. Edn.: April):75-85
© The Author(s). 2013
Received: 03/03/2013 | Accepted: 14/04/2013 | Published: 30/04/2013
Abstract

Expression of Nrf2 in macrophage RAW264.7 induced by serum of septic rat infected with endotoxin and bacteria. Experiments were divided into animal model and cell model. For the animal model, the rats (n=45) were divided into normal control group, LPS group, Pure Burn group, Pseudomonas group, and Staphylococcus group. Rats of all groups were made 40 %total body surface area (TBSA) full thickness scald burn except normal control group. Lethal dose of Staphylococcus aureus, Pseudomonas aeruginosa and LPS were injected into burnt rat peritoneum according to the group respectively. Rats of all groups were killed by cardiac puncture method at 8 hours following burn and bloods collected were centrifuged to collect serum. Macrophage RAW264.7 cells were interfered with the serum separately. RT-PCR and Western blot were performed to study the expression of Nrf2. Nrf2 was prominently expressed in both experiments in all groups compared to normal control. In RT-PCR, Nrf2 mRNA expression level was the highest in the Pure Burn group (p<0.01) which was followed by the LPS group (p<0.01), Pseudomonas group (p<0.01) and Staphylococcus group (p<0.01) respectively. In the Western blot, Nrf2 protein expression was the highest in the LPS group (p<0.01) which was followed by the pure burn group (p<0.01), Pseudomonas group and staphylococcus group (p<0.05) respectively. The expression of Nrf2 indicates that different degree of endogenous anti oxidative system, host inflammatory response and its defense mechanism is active against oxidative stress inside the macrophage RAW264.7 interfered by different pathogens.

Keywords

Nrf2; RAW264.7, Sepsis; LPS, Oxidative stress injury, Endogenous antioxidant system

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