A research on the influence of two herbal concoctions on Toll-like receptor signal pathways of influenza virus induced pneumonia in mice / 中华危重病急救医学

ABSTRACT

Objective To investigate the effect of Shufeng Xuanfei and Jiebiao Qingli concoctions on Toll-like receptor (TLR) signal pathway of pneumonia infected with influenza virus in mice.Methods The pneumonia model was reproduced by nasal dropping of influenza virus A in mice.The mice were randomly divided into nine groupsnormal group (C),model group (M),tamiflu group (D),Shufeng Xuanfei low-dose (SL),medium-dose (SM) and high-dose (SH) groups,Jiebiao Qingli low-dose (JL),medium-dose (JM) and high-dose (JH) groups,each n =12.Two hours after model-reproduction,the mice in C group and M group received distilled water by gavage.The mice in D group received 2.5 g· mL-1· d-1 oseltamivirphosphate.Shufeng Xuanfei formula in doses of 3.76,1.88,0.94 g· kg1 · d-1 were respectively administered to SH,SM and SL groups by gavage,Jiebiao Qingli formula in doses of 4.37,2.18,1.09 g ·kg-1 ·d-1 was given to JH,JM and JL groups by gavage,respectively.Each group was in equal dose of 0.2 mL daily over a 4-day period.Total RNA was extracted in each group.Then gene chips were used to screen these RNA samples.Some genes that were involved in TLR signal pathways were selected.These candidate genes were verified by real-time reverse transcription-polymerase chain reaction (RT-PCR).Results TLR7,MYD88,CCLS,IFNB1,IL6,IL12a,NFKBIA and IKBKB were up-regulated in model group compared with control group.Compared with model group,down-regulated genes in medium-dose,low-dose Shufeng Xuanfei formula and medium-dose Jiebiao Qingli formula included TLR3,TLR7,MYD88,CCL5,IFNB1,IL6,IL12a,NFKBIA and IKBKB (log2 signal intensity of SM,/M in medium-dose Shufeng Xuanfei formula group were-1.24,-2.02,-1.36,-1.95,-0.63,-1.33,-3.50,-1.33,-1.33,log2 signal intensity of SI/M in low-dose Shufeng Xuanfei group were-1.07,-2.43,-2.63,-2.30,-5.09,-3.19,-3.53,-1.95,-1.95,log2 signal intensity of JM/M in medium-dose Jiebiao Qingli formula group were -1.78,-0.55,-1.35,-1.47,-1.65,-2.03,-3.02,-1.57,-1.57,respectively).The results suggested that the effect of Shufeng Xuanfei formula was better than that of Jiebiao Qingli formula.By RT-PCR,compared with model group,low-dose,medium-dose and high-dose groups of Shufeng Xuanfei formula,medium-dose and high-dose groups of Jiebiao Qingli formula,and tamiflu group,significant decrease in TLR7,nuclear factor-κB (NF-κB),myeloid differential protein-88 (MyD88) mRNA expression were found.Medium-dose and low-dose Shufeng Xuanfei formula group (TLR7 mRNA3.6 ±0.3,3.5 ± 1.2 vs.7.4 ± 1.6,NF-κB mRNA1.1 ±0.2,1.0 ±0.2 vs.2.2 ±0.4; MyD88mRNA1.4 ± 0.4,1.0 ± 0.3 vs.3.4 ± 0.9,all P＜0.01) and medium-dose Jiebiao Qingli formula group (TLR7 mRNA4.9 ± 0.3 vs.7.4 ± 1.6,NF-κB aRNA1.3 ± 0.7 vs.2.2 ± 0.4,MyD88 mRNA1.6 ± 0.8 vs.3.4 ± 0.9,P＜0.05 or P＜ 0.01) were shown statistically significant decreases compared with the model group.Conclusions Medium-dose and low-dose Shufeng Xuanfei formula and medium-dose Jiebiao Qingli formula can inhibit the inflammatory reaction induced by influenza virus by down-regulating the NF-κB through TLR signal pathways dependent on MyD88.The regulation of Shufeng Xuanfci formula in TLR signal pathways was superior to that of Jiebiao Qingh formula.