扶正利肺汤对老年社区获得性肺炎患者中性粒细胞TLRs及MyD88 mRNA表达研究

目的观察扶正利肺汤对老年社区获得性肺炎患者外周血中性粒细胞Toll样受体（TLRs）2、4，髓样分化因子88（MyD88）mRNA表达及对细胞因子的调节作用和临床疗效。方法以老年肺炎患者（中西医结合治疗组A，n=38，抗生素治疗组B，n=32）和对照者C组（n=25）作为研究对象。分离外周血中性粒细胞，提取中性粒细胞mRNA，实时定量．PCR检测各组TLR2，TLR4及MyD88mRNA表达；采用ELISA检测外周血上清肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）浓度并观察临床表现。结果对照组外周血中性粒细胞有基础的TLR2、TLR4和MyD88mRNA表达和外周血TNF—α、IL-6产生，分别是0．812±0．227，0．021±0，017，0．948±0，082和（9．520±3．009）pg／ml，（28．820±8．172）pg／ml；而老年肺炎（A组、B组）患者以上指标均显著升高，A组分别是1．096±0．181，0．0394-0．023，1．031±0．088和（25．810±6．737）pg／ml，（65．660±21．296）pg／mhB组分别是1．061±0．226，0．037±0．022，1．029±0．072和（26．160±7．349）pg／ml，（65．960±19．586）pg／ml，较对照组比较差异有统计学意义（P〈0．05）。两组治疗后均使TLR2、TLR4和Myd88mRNA相对表达量下降，虽然无统计学差异，但A组治疗后TLR2mRNA和TLR4mRNA降低趋势更明显，P均〈O．10；两组（A组、B组）治疗后TNF。a和IL．6浓度均显著下降，TNF—α分别为（15．050±3．925）pg／ml，（16．250±3．320）pg／ml，IL-6分别为（39．900±1．634）pg／ml，（40．610±1．836）pg／ml，治疗前后比较差异有统计学意义，分别为t=4．254、3．816、3．734、4．007，P=0．002、0．004、O．005、0.003；治疗后两组比较差异无统计学意义（P=0．470，P=0．372）；但A组发热持续时间显著缩短，差异有统计学意义（f=5．018，P----0．001）；肺部炎症吸收更明显，差异有统计学意义（P=4．119，P=0．042）；同时抗生素治疗时间亦缩短，差异亦有统计学意义（f=3．003，P=0．008）。结论中性粒细胞TLR2、TLR4-Myt）88信号传导通路在老年社区获得性肺炎发生发展中起一定作用。虽然扶正利肺汤单独治疗尚不能影响患者中性粒细胞TLR2、TLR4-Myd88mRNA的表达及细胞因子的释放，但在改善症状、肺部炎症吸收及减少抗生素使用均有益处，提示合理有效抗生素治疗协同扶正利肺汤治疗可能是老年人肺炎治疗新的启示。

The role of FuZhengLiFei decoction in expression of TLRs and MyD88 mRNA of ncutrophil from community acquired pneumonia in the elderly

Objective To investigate whether FuZhengLiFei decoction could have a role in regulating expression of Toll like receptors （TLRs） 2, 4 and myeloid differentiation factor 88 （MyD88） mRNA of peripheral neutrophils and cytokones and clinical effect in olderly patients from pneumonia. Methods Elderly patients with pneumonia （treatment group A of combination of Chinese and Western medicine, n=38; treatment group B of antibiotics, n=32） and control group C （n----25） were selected as research subjects. Neutrophils isolated from patients and controls and neutrophils mRNA extracted. Expression of TLR2, TLR4 and MyD88 mRNA were measured by Real-time PCR. Cytokines （TNF-a and IL-6） in serum were also tested by ELISA and clinical manifestations were observed. Results Peripheral neutrophils of control group have a foundation ofTLR2, TLR4 and MyD88 mRNA expression and production of TNF-a, IL-6, the results were 0.812±0.227, 0.021＋±0.017, 0.948±0.082 and （9.520±3.009） pg/ml, （28.820±8.172） pg/ml respectively and elderly patients with pneumonia were significantly elevated above indicators, group A were 1.096±0.181, 0.039±0.023, 1.031±0.088 and （25.810±6.737）pg/ml, （65.660±21.296） pg/ml respectively and group B were 1.061±0.226, 0.037±0.022, 1.029±0.072 and （26.160±7.349）pg/ml, （65.960± 19.586） pg/ml. Compared with control group, difference was statistically significant （P values were less than 0.05）. After treatment, TLR2, TLR4 and MyD88 mRNA relative expression were declined in both of treatment group, but no statistically significant difference and a decreasing trends was more obvious in group A （P values were less than 0.1）. TNF-a and IL-6 levels were significantly decreased after both treatment（A, B）, they were （15.050±3.925）, （16.250±3.320）pg/rnl and （39.900±1.634） pg/ml, （40.610±1.836） pg/ml respectively （t=4.254, 3.816, 3.734, 4.007, P=0.002, 0.004, 0.005, 0.003）, but compared with no significant difference after both treatment in the concentration of reducing （P= 0.47, P= 0.727）. Group A of duration of fever and antibiotic usage time were more shorter than group B （t=5.018, P=0.001; t= 3.003, P=0.008） and the inflammation in the lung was also significantly improved （X2=4.119, P=0.042）. Conclusion These data suggest that neutrophil TLR2, TLR4 and MyD88 signaling contribute to the development of community acquired pneumonia in the elderly. There are no significant effect of FuZhengLiFei decoction on the expression of Toll like receptors（TLRs） 2, 4 and myeloid differentiation factor 88（MyD88） mRNA of peripheral neutrophils and cytokones from pneumonia in the olderly, but it is beneficial to improving clinical manifestation and absorption of inflammation of lung and reducing use of antibiotics. Suggesting that collaboration with antibiotic and FuZhengLiFei decoction may be a new arousing for treatment of pneumonia in the olderly.