摘要
目的:研究流感病毒性肺炎小鼠肺组织Toll样受体Ⅲ(TLR3)、促分裂原活化蛋白激酶13(P38)、促分裂原活化蛋白激酶8(JNK)表达及疏风宣肺方和解表清里方的调控作用。方法:制备甲型流感病毒性肺炎小鼠模型,随机分为空白组、肺炎模型组、奥司他韦对照组、疏风宣肺方大、中、小剂量组,解表清里方大、中、小剂量组。提取总RNA,采用基因芯片Pathway分析,挑选参与MAPK信号传导通路相关的靶基因。同时,通过实时荧光定量PCR(qRT-PCR)和Western blot技术在mRNA和蛋白表达水平对TLR3、P38、JNK的表达进行验证。结果:肺炎模型组差异表达基因Tlr3、Mapk8、Mapk13明显上调,较空白组差异显著。疏风宣肺方中小剂量和解表清里中剂量组对差异表达基因Tlr3、Mapk8、Mapk13表达有显著的下调作用。qRT-PCR和Western blot法结果显示,肺炎模型组TLR3、JNK、P38的mRNA和蛋白表达较空白组显著升高(P<0.05,P<0.01)。疏风宣肺方中、小剂量组和解表清里方中剂量组对TLR3、JNK、P38的mRNA和蛋白有显著抑制(P<0.05,P<0.01)。且疏风宣肺方的治疗作用存在量效关系,剂量越小,疗效越好。其治疗效果比较如下:疏风宣肺方小剂量组>解表清里方中剂量组>疏风宣肺方中剂量组>解表清里方小剂量组。该结果和基因芯片表达的结果相符。结论:疏风宣肺方和解表清里方可能通过抑制以TLR3介导MAPK信号通路中JNK和P38过度活化来拮抗肺组织炎症损伤和细胞凋亡,从而发挥抗病毒作用。
Objective： To study the regulatory effects of Shufeng Xuanfei Formula （SFCF） and Jiebiao Qingli Formula （JBQL） on expression of TLR3, P38 and JNK in lung of mice with influenza viral pneumonia. Methods： Mice model with influenza viral pneumonia was established, then the mice were randomly divided into nine group： control group, model group, osetamivir group, SFXF groups （high, middle, low dosage group） and JBQL groups （high, middle, low dosage group）. Total RNA was extracted, then pathway was analyzed using gene chip, and the target genes participated in the MARK signal transduction pathway were selected. The RNA and protein expression of TLR3, P38 and JNK was detected with qRT-PCR and western blot. Results： Compared with the control group, the differential expression gene of Tlr3, Mapk8 and Mapkl3 was increased significantly in the model group, and low and middle dosage of XFSF and middle dosage of JBQL could down-regulate the expression of Tlr3, Mapk8 and Mapkl3. Results of qRT-PCR and western blot showed that the protein and mRNA expression of TLR3, JNK and P38 of the model group was higher than that of the control group （P〈0.05, P〈0.01）. Low and middle dosage of XFSF and middle dosage of JBQL could inhibit the mRNA and protein expression of TLR3, JNK and P38 significantly （P〈0.05, P〈0.01）. There was a concentration-response relationship in the therapeutic effect of SFXF formula, the curative effect was better with dose reduction of SFXF formula. The treatment effect was as follows： low dosage of SFXF〉 middle dosage of JBQL〉 middle dosage of SFXF〉 low dosage of JBQL, and this result agreed with the result of gene chip. Conclusion： SFXF and JBQL formulas could fight against the inflammatory damage of lung tissue and cell apoptosis to play a role of antivirus by inhibiting the excessive activation of JNK and P38 in MAPK signal pathway mediated by TLR3.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2014年第11期3563-3566,共4页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81173371)
北京中医药大学校级自主课题(No.2013-JYBZZXS-016)~~
