long-TSLP和short-TSLP调控肺成纤维细胞有氧糖酵解在哮喘气道重塑中的作用和机制研究

Thymic stromal lymphopoietin (TSLP) plays an important role in airway chronic inflammation and remodeling of asthma. Recent reports that the long-TSLP, a subtype of TSLP, could promoted proliferation and extracellular matrix (ECM) production, but the effects of long-TSLP could be inhibited by short-TSLP. However, the mechanism of long-TSLP and short-TSLP in regulating proliferation and ECM of lung fibroblast is unclear. The preliminary study found that aerobic glycolysis was up-regulation by long-TSLP. Therefore, this project would used CRISPR / Cas9 gene knockout, immunofluorescence and in situ hybridization to investigate whether long-TSLP promoted proliferation and ECM through up-regulating aerobic glycolysis of lung fibroblast to participate in the development of airway remodeling. And to further illustrate the specific mechanism of long-TSLP inducing aerobic glycolysis of lung fibroblast by TSLPR/IL-7Rα-STAT3/5-HIF-1α and the effects of short-TSLP in that. These results would provide new theory basis for treatment of asthma.

胸腺基质淋巴细胞生成素（TSLP）在哮喘气道慢性炎症和重塑中有重要作用。新近研究发现，TSLP的亚型长链TSLP（long-TSLP）可促进肺成纤维细胞增殖及合成细胞外基质（ECM），而短链TSLP（short-TSLP）能抑制long-TSLP的作用。然而，long-TSLP和short-TSLP调控肺成纤维细胞增殖及合成ECM的机制尚不清楚。本课题组前期研究工作发现，long-TSLP可促进肺成纤维细胞有氧糖酵解。因此，本课题拟运用CRISPR/Cas9基因敲除、免疫荧光、原位杂交等技术，观察是否long-TSLP通过诱导肺成纤维细胞有氧糖酵解，诱导其增殖及合成ECM，促进气道重塑的发生发展；并进一步探讨long-TSLP通过 TSLPR/IL-7Rα-STAT3/5-HIF-1α上调肺成纤维细胞有氧糖酵解的机制，及short-TSLP在其中的作用。为哮喘气道重塑防治提供新的策略。

背景：胸腺基质淋巴细胞生成素（TSLP）在哮喘气道慢性炎症和重塑中有重要作用。新近研究发现，TSLP的亚型长链TSLP（long-TSLP，lTSLP）可促进肺成纤维细胞增殖及合成细胞外基质（ECM），而短链TSLP（short-TSLP，sTSLP）能抑制long-TSLP的作用。然而，long-TSLP和short-TSLP调控肺成纤维细胞增殖及合成ECM的机制尚不清楚。.目的：阐明有氧糖酵解在哮喘气道上皮下肺成纤维细胞增殖、转化为肌成纤维细胞和ECM合成中的作用，及lTSLP和sTSLP调控肺成纤维细胞有氧糖酵解参与气道重塑的作用和机制。.结果：lTSLP可以通过TSLPR/IL-7R受体复合物激活STAT5促进气道上皮细胞有氧糖酵解，进而增加炎症因子释放；另一方面，lTSLP能直接促进气道上皮下肺成纤维细胞有氧糖酵解而激活成纤维细胞，从而参与气道重塑。而sTSLP能占据TSLPR/IL-7R受体，减轻lTSLP活化的JAK1/2-STAT5信号通路，下调miR-223水平，促进VHL介导的HIF-1α羟基化水解，抑制上皮细胞有氧糖酵解和炎症因子释放；同时sTSLP能抑制lTSLP诱导的肺成纤维细胞有氧糖酵解和活化，减轻哮喘气道重塑。.意义：阐明lTSLP在哮喘慢性气道炎症和气道重塑中的作用机理，并初步揭示了sTSLP改善哮喘慢性气道炎症和气道重塑的机制。为进一步开发sTSLP作为治疗哮喘药物的可能性提供了理论依据。