Liu Qi, Wang Jianguo, Guo Yu, Jia Lu, Yuan Haijun, Yang Wanfang, Wen Yangxuan
Shanxi University of Traditional Chinese Medicine, Jinzhong 030619, Shanxi, China.
Shanxi Hospital of Integrated Traditional and Western Medicine, Taiyuan 030013, Shanxi, China. Corresponding author: Liu Qi, Email:
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2020 Feb;32(2):183-187. doi: 10.3760/cma.j.cn121430-20190603-00034.
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.
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).
Compared with group N, the differential gene expressions of IL-1, IL-8 and ICAM-1 in group M were significantly up-regulated [log2 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 were -1.91, -1.85, -0.88; log2 were -3.10, -1.74, -1.84; log2 were -1.89, -1.39, -0.53; log2 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): 4.63±0.24 vs. 1.01±0.13, IL-8 (2): 6.28±0.13 vs. 1.02±0.09, ICAM-1 (2): 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): 2.12±0.32, 1.71±0.07, 2.05±0.16, 1.66±0.13 vs. 4.63±0.24; IL-8 (2): 3.89±0.13, 2.08±0.19, 2.98±0.20, 2.02±0.12 vs. 6.28±0.13; ICAM-1 (2): 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.
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.
采用基因芯片技术分析疏风宣肺解毒方对流感病毒感染小鼠炎症相关细胞因子全表达谱的差异基因表达。
将雄性ICR小鼠按随机数字表法分为正常组(N组)、流感病毒肺炎模型组(M组)、磷酸奥司他韦对照组(C组)及疏风宣肺解毒方高、中、低剂量组(SH、SM、SL组),每组10只。通过滴鼻流感病毒株FM1(0.05 mL)建立小鼠流感病毒肺炎模型;N组滴注0.05 mL生理盐水。SH、SM和SL组在鼻内感染2小时后给予疏风宣肺解毒方(药物浓度分别约为3.8、1.9和1.0 kg/L),每天0.2 mL,连续4天;C组磷酸奥司他韦剂量为2.5 kg/L;N组和M组给予蒸馏水。第5天,采集小鼠全肺,提取肺组织总RNA,与小鼠全基因表达谱芯片杂交后进行检测。筛选参与炎症途径的细胞因子差异表达基因。计算每组芯片探针信号与M组的强度表达比值,以P < 0.05且log2比值>1定义为上调基因,P < 0.05且log2比值< -1定义为下调基因。采用逆转录-聚合酶链反应(RT-PCR)检测白细胞介素(IL-1、IL-8)和细胞间黏附分子-1(ICAM-1)的mRNA表达。
与N组比较,M组IL-1、IL-8和ICAM-1的差异基因表达显著上调[log2分别为2.62、2.07、1.41,均P < 0.05]。与M组比较,SH、SM、SL和C组IL-1、IL-8、ICAM-1的基因表达显著下调[log2分别为-1.91、-1.85、-0.88;log2分别为-3.10、-1.74、-1.84;log2分别为-1.89、-1.39、-0.53;log2分别为-2.46、-1.52、-1.44,均P < 0.05]。RT-PCR显示,M组IL-1、IL-8和ICAM-1的mRNA表达显著高于N组[IL-1(2):4.63±0.24对1.01±0.13,IL-8(2):6.28±0.13对1.02±0.09,ICAM-1(2):2.90±0.18对1.02±0.12,均P < 0.05]。SH、SM、SL和C组IL-1、IL-8、ICAM-1的mRNA表达低于M组[IL-1(2):2.12±0.32、1.71±0.07、2.05±0.16、1.66±0.13对4.63±0.24;IL-8(2):3.89±0.13、2.08±0.19、2.98±0.20、2.02±0.12对6.28±0.1