基于上皮细胞-间充质细胞转分化(EMT)理论的艾灸配合化纤Ⅳ号方对实验大鼠Collagen Type Ⅲ和PDGF干预作用实验研究

Intervention of Moxibustion and Huaxian Ⅳ Formula on Rats' Collagen Type Ⅲ and PDGF Based on EMT

目的:基于上皮细胞-间充质细胞转分化(EMT)学说观察化纤Ⅳ号方、艾灸以及二者相配合治疗肺纤维化大鼠Collagen TypeⅢ(Ⅲ-C)和PDGF的变化,探讨其治疗效应及生物学机制。方法:将鼠龄约为6周的SD大鼠随机分为空白组、模型组、化纤Ⅳ号方组、艾灸组、化纤Ⅳ号方与艾灸配合治疗组(简称为"灸药组"),治疗30 d后处死观察其肺组织病理改变,并检测其Collagen TypeⅢ、PDGF的基因和蛋白表达情况。结果:实时荧光定量结果显示:与空白组相比,各组Ⅲ-C和PDGF m RNA表达增高(P<0.05)。与模型组相比,各组的Ⅲ-C和PDGF m RNA表达有明显降低(P<0.01)。而各组中,灸药组疗效最明显,Ⅲ-C和PDGF的表达最低。蛋白免疫印迹法检测结果显示:与模型组相比各组的Ⅲ-C蛋白表达有差异。结论:1艾灸、化纤Ⅳ号方均可减轻博莱霉素诱导肺纤维化大鼠的肺纤维化程度。2艾灸配合化纤Ⅳ号方可减轻博莱霉素诱导肺纤维化大鼠的肺纤维化程度,且其效果优于单用艾灸或单用化纤Ⅳ号方。3艾灸、化纤Ⅳ号方及其二者配合使用不同程度阻抑博莱霉素诱导肺纤维化大鼠肺纤维化进程的效应机制,可能与通过调控其EMT过程中的Ⅲ-C和PDGF表达环节紧密相关。

Objective： To observe the changes of collagen type Ⅲ and PDGF of lung fibrosis rats treated by moxibustion and Huaxian Ⅳ Formula or combination of both and discuss the mechanism. Methods： SD rats aged 6 weeks were randomly divided into blank group,model group,Huaxian Ⅳ Formula group,moxibustion group and the combination group. After 30 d treatment,the rats were killed to observe the lung tissues changes. We tested collagen type Ⅲ and PDGF expressions. Results： Compared with the blank group,each group＇s collagen type Ⅲ and PDGF m RNA expressions were increased（ P 〈0. 05）. Compared with the model group,each group＇s collagen type Ⅲ and PDGF m RNA expressions were decreased obviously（ P 〈0. 01）. The combination group had the most obvious effect collagen type Ⅲ and PDGF m RNA expressions were the lowest. Western blot study showed that compared with the model group,the other groups＇ Ⅲ-C expressions had significance. Conclusion： ①Both moxibustion and Huaxian ⅣFormula can relieve the lung fibrosis due to bleomycin. ② The combination of moxibustion and Huaxian Ⅳ Formula has better effect than single use. ③. The mechanism may be regulating Ⅲ-C and PDGF during EMT.