淫羊藿苷对慢性阻塞性肺疾病模型的抗炎和抗氧化作用

Anti-inflammatory and Antioxidant Effects of Icariin on Chronic Obstructive Pulmonary Disease Model

目的探讨淫羊藿苷对香烟烟雾和脂多糖(LPS)刺激的大鼠肺泡巨噬细胞(NR8383)和大鼠模型炎症的影响。方法以LPS刺激NR8383细胞系建立细胞炎症模型,以香烟烟雾和LPS联合刺激SD大鼠建立慢性阻塞性肺疾病(COPD)模型,采用淫羊藿苷干预。采用MTT检测细胞活力;采用微板酶标法和分光光度计检测细胞培养上清和血清中NO,i NOS和SOD含量;采用流式细胞技术检测外周血CD4+和CD8+T细胞含量;采用病理切片HE染色观察肺组织病理变化。结果体外研究表明:淫羊藿苷80μM时显示出最接近空白组的细胞活力,淫羊藿苷10、20、40、80和160μM均显示出抑制NO和i NOS的释放(P>0.05),而淫羊藿苷5、10、20、40、80和160μM均提高了SOD的活力(P<0.05)。体内研究表明:淫羊藿苷干预之后的大鼠血清NO和i NOS释放均明显降低,SOD活力显著升高(P<0.05),CD8+T细胞比例比例下降(P<0.05),CD4+/CD8+T细胞的比值升高(P<0.05),减轻大鼠肺部炎症细胞浸润。结论淫羊藿苷是一种安全有效的天然抗炎抗氧化药物,可能通过改变NO、i NOS、SOD和T淋巴细胞的含量来实现疗效。

Objective To investigate the anti - inflammatory and antioxidant effects of icariin on cigarette smoke （CS） - exposed/LPS -induced NR8383 cell and rats. Methods Inflammation model in vitro was established by stimulating macrophage cell line NR8383 cell with LPS;and COPD rat model in vivo was established by stimulating SD rats with cigarette smoke and LPS. Both models were treated with icariin, and the cell viability was detected with MTT;NO, iNOS and SOD in cell culture medium and serum were detected by ELISA and spectrophotometry ; CD4 ^＋ and CD8 ^＋ T lymphocytes were enumerated in peripheral blood by flow cytometry, and pulmonary pathological changes were observed by pathological section as well. Results icariin at a concentration of 80 μM did not interfere with the survival of NR8383 macrophages. In vitro studies showed that 10μM , 20μM , 40μM , 80μM and 160μM icariin effectively reduced the levels of NO and iNOS （ P 〈 0. 05）. 5 μM, 10μM , 20μM , 40μM , 80μM and 160μM icariin significantly increased the activity of SOD（ P 〈 0. 05 ）. In vivo studies showed icariin inhibited the expression the production of NO and iNOS , and elevated the activity of SOD in serum significantly （P 〈 0. 05 ）. icariin treatment significantly lowered the percentage of CD8 ^＋ T cells （ P 〈 0.05 ）, and reversed the ratio of CD4^＋ /CD8 ^＋ T cells （ P 〈 0. 05）. Icariin alleviated pulmonary inflammatory cell infiltration. Conclusion Icariin is a safe and effective natural anti - inflammatory drug. Icariin exerts therapeutic effects by changing the production of NO, iNOS, SOD and T lymphocytes.