疏风宣肺和解表清里方药对流感病毒性肺炎小鼠Toll样受体通路影响的研究
소풍선폐화해표청리방약대류감병독성폐염소서Toll양수체통로영향적연구
A research on the influence of two herbal concoctions on Toll-like receptor signal pathways of influenza virus induced pneumonia in mice
  • 万方数据
    中华危重病急救医学 중화위중병급구의학
    Chinese Critical Care Medicine
    2014年 5期 321-324 ,共4页

    刘琪%王建国%马彦平%顾立刚

    류기%왕건국%마언평%고립강

    疏风宣肺方%解表清里方%流感病毒性肺炎%Toll样受体%基因芯片%小鼠 疏風宣肺方%解錶清裏方%流感病毒性肺炎%Toll樣受體%基因芯片%小鼠
    소풍선폐방%해표청리방%류감병독성폐염%Toll양수체%기인심편%소서
    Shufeng Xuanfei formula%Jiebiao Qingli formula%Pneumonia infected with influenza virus%Toll-like receptor%Gene chip%Mouse
    目的 观察疏风宣肺、解表清里方对流感病毒亚甲型肺适应株FM1感染小鼠肺组织细胞Toll样受体(TLR)信号转导通路的影响.方法 制备小鼠甲型流感病毒性肺炎模型,按随机数字表法分为对照组(C),模型组(M),达菲组(D),疏风宣肺方高、中、低剂量组(SH、SM、SL),解表清里方高、中、低剂量组(JH、JM、JL),每组12只.制模后2h,对照组、模型组均灌胃蒸馏水,达菲组灌胃达菲2.5 g·mL-1·-1,不同剂量方药组分别灌胃疏风宣肺方颗粒剂3.76、1.88、0.94 g·kg-1·d-1和解表清里方颗粒剂4.37、2.18、1.09 g·kg-1·d-1,各组均0.2 mL/d,连用4d.提取肺组织总RNA,采用基因组芯片筛选出与TLR通路相关的差异表达基因,并用实时荧光定量逆转录-聚合酶链反应(RT-PCR)对部分基因进行验证.结果 与对照组比较,模型组差异表达基因TLR7、MYD88、CCL5、IFNB1、IL6、IL12a、NFKBIA、IKBKB明显上调.与模型组比较,疏风宣肺方中、低剂量组和解表清里方中剂量组差异表达基因TLR3、TLR7、MYD88、CCL5、IFNB1、IL6、IL12a、NFKBIA、IKBKB明显下调[疏风宣肺方中剂量组log2(SM组/M组信号强度)分别为-1.24、-2.02、-1.36、-1.95、-0.63、-1.33、-3.50、-1.33、-1.33,疏风宣肺方低剂量组log2(SL组/M组信号强度)分别为-1.07、-2.43、-2.63、-2.30、-5.09、-3.19、-3.53、-1.95、-1.95;解表清里方中剂量组log2(JM组/M组信号强度)分别为-1.78、-0.55、-1.35、-1.47、-1.65、-2.03、-3.02、-1.57、-1.57],提示疏风宣肺方治疗效果比解表清里方好.RT-PCR结果显示,与模型组比较,疏风宣肺方高、中、低剂量组和解表清里方高、中剂量组及达菲组对TLR7、核转录因子-KB(NF-κB)、髓样分化抗原88(MyD88)的mRNA表达均有抑制作用,其中疏风宣肺方中、低剂量组(TLR7 mRNA:3.6±0.3、3.5±1.2比7.4±1.6; NF-κBmRNA:1.1±0.2、1.0±0.2比2.2±0.4;MyD88 mRNA:1.4±0.4、1.0±0.3比3.4±0.9,均P＜0.01)和解表清里方中剂量组(TLR7 mRNA:4.9±0.3比7.4± 1.6; NF-κB mRNA:1.3 ±0.7比2.2±0.4; MyD88mRNA:1.6±0.8比3.4±0.9,P＜0.05或P＜0.01)作用较为明显.结论 中、低剂量疏风宣肺方和中剂量解表清里方通过调控MyD88依赖的TLR信号转导通路下调NF-κB表达,从而治疗流感;疏风宣肺方药疗效较好.
    목적 관찰소풍선폐、해표청리방대류감병독아갑형폐괄응주FM1감염소서폐조직세포Toll양수체(TLR)신호전도통로적영향.방법 제비소서갑형류감병독성폐염모형,안수궤수자표법분위대조조(C),모형조(M),체비조(D),소풍선폐방고、중、저제량조(SH、SM、SL),해표청리방고、중、저제량조(JH、JM、JL),매조12지.제모후2h,대조조、모형조균관위증류수,체비조관위체비2.5 g·mL-1·-1,불동제량방약조분별관위소풍선폐방과립제3.76、1.88、0.94 g·kg-1·d-1화해표청리방과립제4.37、2.18、1.09 g·kg-1·d-1,각조균0.2 mL/d,련용4d.제취폐조직총RNA,채용기인조심편사선출여TLR통로상관적차이표체기인,병용실시형광정량역전록-취합매련반응(RT-PCR)대부분기인진행험증.결과 여대조조비교,모형조차이표체기인TLR7、MYD88、CCL5、IFNB1、IL6、IL12a、NFKBIA、IKBKB명현상조.여모형조비교,소풍선폐방중、저제량조화해표청리방중제량조차이표체기인TLR3、TLR7、MYD88、CCL5、IFNB1、IL6、IL12a、NFKBIA、IKBKB명현하조[소풍선폐방중제량조log2(SM조/M조신호강도)분별위-1.24、-2.02、-1.36、-1.95、-0.63、-1.33、-3.50、-1.33、-1.33,소풍선폐방저제량조log2(SL조/M조신호강도)분별위-1.07、-2.43、-2.63、-2.30、-5.09、-3.19、-3.53、-1.95、-1.95;해표청리방중제량조log2(JM조/M조신호강도)분별위-1.78、-0.55、-1.35、-1.47、-1.65、-2.03、-3.02、-1.57、-1.57],제시소풍선폐방치료효과비해표청리방호.RT-PCR결과현시,여모형조비교,소풍선폐방고、중、저제량조화해표청리방고、중제량조급체비조대TLR7、핵전록인자-KB(NF-κB)、수양분화항원88(MyD88)적mRNA표체균유억제작용,기중소풍선폐방중、저제량조(TLR7 mRNA:3.6±0.3、3.5±1.2비7.4±1.6; NF-κBmRNA:1.1±0.2、1.0±0.2비2.2±0.4;MyD88 mRNA:1.4±0.4、1.0±0.3비3.4±0.9,균P＜0.01)화해표청리방중제량조(TLR7 mRNA:4.9±0.3비7.4± 1.6; NF-κB mRNA:1.3 ±0.7비2.2±0.4; MyD88mRNA:1.6±0.8비3.4±0.9,P＜0.05혹P＜0.01)작용교위명현.결론 중、저제량소풍선폐방화중제량해표청리방통과조공MyD88의뢰적TLR신호전도통로하조NF-κB표체,종이치료류감;소풍선폐방약료효교호.
    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 groups:normal 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 mRNA:3.6 ±0.3,3.5 ± 1.2 vs.7.4 ± 1.6,NF-κB mRNA:1.1 ±0.2,1.0 ±0.2 vs.2.2 ±0.4; MyD88mRNA:1.4 ± 0.4,1.0 ± 0.3 vs.3.4 ± 0.9,all P＜0.01) and medium-dose Jiebiao Qingli formula group (TLR7 mRNA:4.9 ± 0.3 vs.7.4 ± 1.6,NF-κB aRNA:1.3 ± 0.7 vs.2.2 ± 0.4,MyD88 mRNA:1.6 ± 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.
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