【摘要】 目的探讨加味过敏煎和玉屏风散通过调节辅助性T细胞17(Th17)/调节性T细胞(Treg)平衡干预PM2.5致肺损伤作用机制。方法 40只SPF级雄性Wistar大鼠随机分为正常对照组、模型组、加味过敏煎组和玉屏风散组。采用气道内滴注法建立PM2.5致肺损伤的动物模型,加味过敏煎组予加味过敏煎颗粒剂(2.10 g/kg)灌胃,玉屏风散组予玉屏风散颗粒剂(1.89 g/kg)灌胃,正常对照组和模型组予等量蒸馏水灌胃;连续28 d。通过HE染色观察肺组织病理学改变,ELISA法检测血清和肺泡灌洗液(BALF)中白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)及BALF中白细胞介素-17(IL-17)和白细胞介素-10(IL-10)的含量,免疫组化法和Western-blot法检测Th17核转录因子视黄酸相关的孤儿受体γt(RORγt)和Treg细胞核转录因子叉头蛋白P3(FoxP3)蛋白的表达,实时荧光定量PCR法检测IL-17、IL-10、RORγt、FoxP3的mRNA表达。结果与正常对照组比较,模型组大鼠肺组织气道上皮层及肺泡壁破坏明显,血清IL-1β和IL-6含量显著增加(P<0.05),BALF中IL-17A含量显著增加,肺组织中RORγt蛋白表达升高,IL-10 mRNA表达降低,RORγt mRNA表达显著升高(P<0.05)。与模型组比较,加味过敏煎组和玉屏风散组肺组织损伤情况减轻,血清中IL-6含量显著减少,肺组织中RORγt蛋白表达降低,FoxP3蛋白表达和IL-10 mRNA表达增加,玉屏风散组血清IL-1β和BALF中IL-17A含量减少(P<0.05)。与加味过敏煎组比较,玉屏风散组各指标差异均无统计学意义(P>0.05)。结论 PM2.5可通过引起Th17/Treg失衡诱发肺损伤,加味过敏煎和玉屏风散可以一定程度改善PM2.5致大鼠病理损害,具有调节PM2.5致大鼠Th17/Treg失衡的作用。更多还原

【Abstract】 Objective To investigate the effect and mechanism of Modified Guomin Decoction(MGMD) and Yupingfeng Powder(YPFP) on lung injury caused by PM2.5 via regulating the balance of T helper 17 cells(Th17)/regulatory T cells(Treg). Methods 40 SPF male Wistar rats were randomly divided into control group, PM2.5 model group, MGMD group and YPFP group(n=10). The PM2.5 animal model was established by intratracheal instilling of PM2.5 suspensions. In MGMD and YPFP groups, rats were administrated MGMD and YPFP via gavage every day respectively; in the other two groups, distilled water were gavaged. All rats were sacrificed after 28 days. At the end of the experiment, BALF and lung tissues were collected. The lung histopathological changes were observed by immunohistochemistry. The expression of IL-6 and TGF-β1 in serum and alveolar lavage fluid(BALF) and the content of IL-17 and IL-10 in BALF were detected by ELISA. The expression of RORγt and FoxP3 was detected by immunohistochemistry and Western Blot. The mRNA expression in IL-17, IL-10, RORγt and FoxP3 was detected by real-time PCR. Results Compared with the control group, in the serum of PM2.5 model group the expression of IL-1β and IL-6 was significantly increased(P<0.05), the content of IL-17 A in BALF was significantly increased(P<0.05), and the IL-10 mRNA in lung tissue was reduced while RORγt mRNA was significantly increased(P<0.01). Compared with the model group, in both MGMD group and YPFP group, the lung tissue injury wasmilder, the expression of IL-6 in serum and RORγt in lung tissue was decreased, the expression of FoxP3 and IL-10 mRNA was increased, and the content of IL-1β in serum and IL-17 A in BALF was decreased in the YPFP group(P<0.05). Compared with the MGMD group, all indicator in the YPFP group were not statistically significant(P>0.05). Conclusion PM2.5 seems to induce lung injury by causing Th17/Treg imbalance, and MGMD and YPFP could possibly improve the pathological damage caused by PM2.5 in rats to a certain extent through restoring Th17/Treg balance.更多还原