加味芍甘附子汤调控PI3K/Akt信号通路减缓类风湿关节炎相关间质性肺病进展的研究

Jiawei Shaogan Fuzi Decoction regulates PI3K/Akt signaling pathway to reduce rheumatoid arthritis related research on the progress of interstitial lung disease

目的 探讨加味芍甘附子汤调控磷脂酰肌醇3激酶/蛋白激酶B(phosphatidylinositol 3 kinase/protein kinase, PI3K/Akt)信号通路控制炎症反应,进而延缓类风湿关节炎相关间质性肺病进展的机制。方法 选用清洁级雌性Wistar大鼠48只,随机分为空白组、模型组、雷公藤组[雷公藤多苷6.25 mg/(kg·d)]、中药大、中、小剂量组[加味芍甘附子汤17.70 g/(kg·d)、8.85 g/(kg·d)、4.43 g/(kg·d)],每组8只。除空白组外,其余五组建立关节炎寒证模型。造模成功后,除空白组、模型组外,其余各组连续4周给与相应药物灌胃治疗。治疗后,观察肺组织病理状态,免疫组化法检测肺组织中PI3K、Akt、核因子κB(nuclear factor kappa-B,NF-κB)蛋白的表达。结果 与空白组相比,模型组大鼠肺组织病理结构大面积破坏,肺泡结构塌陷,肺泡间隙充血水肿,肺泡壁增厚,出现大量纤维瘢痕。与模型组相比,中药中、大剂量组及雷公藤组肺泡结构较为清晰,伴有轻微炎症反应及少量纤维增生。肺泡炎评分显示,模型组与空白组炎症反应有明显差异(P<0.01);中药中、大剂量组与模型组相比,肺泡炎症和纤维化有显著下降,差异有统计学意义(P<0.01);雷公藤组与模型组相比也有明显差异(P<0.01)。与空白组相比,模型组中PI3K、Akt、NF-κB蛋白的表达明显增多。各治疗组PI3K、Akt、NF-κB蛋白的表达与模型组相比均不同程度降低。荧光光密度值对比结果显示,模型组吸光度值较空白组明显升高,差异具有统计学意义(P<0.05);各治疗组较模型组吸光度值均有降低。与模型组相比,PI3K、NF-κB在中药中、大剂量组与雷公藤组中降低更明显(P<0.05)。结论 加味芍甘附子汤通过调节PI3K/Akt信号通路进而干预其下游NF-κB信号通路抑制炎症反应,可能是减少肺泡炎及肺纤维化的重要机制。

Objective To explore the mechanism of Jiawei Shaogan Fuzi Decoction in regulating PI3K/Akt signaling pathway to control inflammatory reaction, and then delaying the progress of interstitial lung disease associated with rheumatoid arthritis. Methods Forty-eight female Wistar rats were randomly divided into the blank group, the model group, the tripterygium wilfordii group [6.25 mg/(kg·d)] and the traditional Chinese medicine group [large, medium and small dosage groups, Jiawei Shaogan Fuzi Decoction 17.70 g/(kg·d), 8.85 g/(kg·d), 4.43 g/(kg·d)], with 8 rats in each group. Except the blank group, rats in the other five groups were established CIA cold syndrome model. After successful modeling, except the blank group and the model group, all other groups were given corresponding drugs for 4 weeks. After treatment, the pathological state of lung tissue was observed, and the expressions of PI3K, Akt and NF-κB protein in lung tissues were detected by immunohistochemistry. Results Compared with the blank group, the pathological structure of the lung tissue in the model group was destroyed in a large area, the alveolar structure collapsed, the alveolar space was congested and edema, the alveolar wall was thickened, and a large number of fibrous scars appeared. Compared with the model group, the alveolar structure of the middle and high dose group of traditional Chinese medicine and tripterygium wilfordii group was clearer, accompanied by slight inflammatory reaction and a small amount of fibrous hyperplasia. Alveolar inflammation score showed that there was a significant difference between the model group and the blank group in inflammatory response(P<0.01). Compared with the model group, the alveolar inflammation and fibrosis in the middle and high dose group of traditional Chinese medicine was decreased significantly(P<0.01), and there was also a significant difference between the tripterygium wilfordii group and the model group(P<0.01). Compared with the blank group, the expressions of PI3K, Akt and NF-κB protein in the model group were increased significantly. Compared with the model group, the expressions of PI3K, Akt and NF-κB protein in each treatment group were decreased in different degrees. The comparison results of fluorescence optical density also showed that the absorbance of the model group was significantly higher than that in the blank group, and the difference was statistically significant(P<0.05). Compared with the model group, the absorbance values of each treatment group were lower, and PI3K and NF-κB in the middle-high dose group and tripterygium wilfordii group were decreased significantly(P<0.05). Conclusion Jiawei Shaogan Fuzi Decoction inhibits inflammation by regulating PI3K/Akt signaling pathway and then intervening its downstream NF-κB signaling pathway, which may be an important mechanism to reduce alveolitis and pulmonary fibrosis.