Effects of Shufeng Xuanfei Jiedu formula for the inflammation-related cytokines in pneumonia mice infected with influenza virus / 中华危重病急救医学

ABSTRACT

Objective:To analyze the differential gene expression of Shufeng Xuanfei Jiedu formula on whole expression profiles of the inflammation-related cytokines in mice infected with influenza virus by the gene chip technology.Methods:Male ICR mice were divided into normal group (N group), influenza virus pneumonia model group (M group), oseltamivir control group (C group) and Shufeng Xuanfei Jiedu formula high, medium and low dose groups (SH, SM, SL groups) according to the random number table method, with 10 mice in each group. A mouse model of influenza virus pneumonia was established by nasal drip of influenza virus strain FM1 (0.05 mL); in group N, 0.05 mL normal saline was used. In SH, SM and SL groups, Shufeng Xuanfei Jiedu formula was prescribed after 2 hours of intranasal infection (drug concentration approximately 3.8, 1.9 and 1.0 kg/L), 0.2 mL once a day for 4 days; in group C, the dosage of oseltamivir was 2.5 kg/L; in group N and group M, distilled water was given. On the 5th day, the whole lung of mice was harvested, and the total RNA of lung tissue was extracted and detected after hybridization with mice whole gene expression spectrum chip. Differential expressed genes of cytokines involved in inflammatory pathways were selected. The intensity expression ratio of the chip probe signal in each group vs. M group was calculated, and P < 0.05 and log2 ratio > 1 were defined as up-regulated genes, while P < 0.05 and log2 ratio < -1 were down-regulated genes. The mRNA expressions of interleukin (IL-1, IL-8) and intercellular adhesion molecule-1 (ICAM-1) were detected by reverse transcription-polymerase chain reaction (RT-PCR). Results:Compared with group N, the differential gene expressions of IL-1, IL-8 and ICAM-1 in group M were significantly up-regulated [log2 (N/M) were 2.62, 2.07, 1.41, respectively, all P < 0.05]. Compared with group M, the gene expressions of IL-1, IL-8, ICAM-1 were significantly down-regulated in SH, SM, SL and C groups [log2 (SH/M) were -1.91, -1.85, -0.88; log2 (SM/M) were -3.10, -1.74, -1.84; log2 (SL/M) were -1.89, -1.39, -0.53; log2 (C/M) were -2.46, -1.52, -1.44, respectively, all P < 0.05]. RT-PCR showed that the mRNA expressions of IL-1, IL-8 and ICAM-1 in group M were significantly higher than those in group N [IL-1 (2 -&Delta;&Delta;CT) 4.63±0.24 vs. 1.01±0.13, IL-8 (2 -&Delta;&Delta;CT) 6.28±0.13 vs. 1.02±0.09, ICAM-1 (2 -&Delta;&Delta;CT) 2.90±0.18 vs. 1.02±0.12, all P < 0.05]. The mRNA expressions of IL-1, IL-8, ICAM-1 in SH, SM, SL and C groups were lower than those in group M [IL-1 (2 -&Delta;&Delta;CT) 2.12±0.32, 1.71±0.07, 2.05±0.16, 1.66±0.13 vs. 4.63±0.24; IL-8 (2 -&Delta;&Delta;CT) 3.89±0.13, 2.08±0.19, 2.98±0.20, 2.02±0.12 vs. 6.28±0.13; ICAM-1 (2 -&Delta;&Delta;CT) 1.72±0.93, 1.34±0.14, 1.53±0.25, 1.17±0.12 vs. 2.90±0.18, all P < 0.05]. There was no significant difference among the SH, SM, SL and C groups. Conclusion:Shufeng Xuanfei Jiedu formula inhibits inflammatory damage in mice after influenza virus infection by down-regulating the expressions of IL-1, IL-8, and ICAM-1 inflammatory cytokine-related genes.