Umbilical cord mesenchymal stem cells inhibited inflammation of bronchial epithelial cells by regulating Hedgehog pathway

Abstract

This study aimed to explore the role and mechanism of umbilical cord mesenchymal stem cells (UCMSCs) in regulating inflammation of bronchial epithelial cells. Transforming growth factor beta-1 (TGF-β1) was used to induce inflammation in human bronchial epithelial cells. Cell proliferation was detected through CCK8 and cell apoptosis was detected by Annexin V and propidium iodide double staining. E-cadherin and α-smooth muscle actin (α-SMA) were detected by immunofluorescence, and tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) in culture medium supernatant were detected by ELISA. The expression of E-cadherin, α-SMA, Sonic hedgehog (Shh), Gli1 and Snail was detected by Western blot analysis. Compared with the control group, bronchial epithelial cells treated with TGF-β1 showed significantly decreased proliferation, increased apoptosis, increased secretion of TNF-α and IL-6, increased expression of α-SMA, Shh, Gli1 and Snail and decreased E-cadherin expression. However, co-culture with UCMSCs inhibited TGF-β1-induced changes in human bronchial epithelial cell proliferation, apoptosis, secretion of TNF-α and IL-6 and activation of the Hedgehog pathway. In conclusion, UCMSCs have protective effects on TGF-β1-induced inflammation in human bronchial epithelial cells by regulating the Hedgehog pathway.

Figure 1.

Identification of human bronchial epithelial cells and hUCMSCs. A) Identification of human bronchial epithelial cells; scale bars: 100 μm. B) Identification of hUCMSCs.

Figure 2.

Effect ofH pathway inhibitor and MSCs on the proliferation and apoptosis of human bronchial epithelial cells. A) CCK8 assay of cell proliferation. B) Flow cytometry analysis of apoptosis. *p<0.05 compared with control group; #p<0.05 compared model group.

Figure 3.

Effect of Hedgehog pathway inhibitor and UCMSCs on the secretion of inflammatory factors in human bronchial epithelial cells. TNF-α and IL-6 in the supernatants were detected by ELISA. *p<0.05 compared with control group; ^#p<0.05 compared model group.

Figure 4.

Effect of Hedgehog pathway inhibitor and UCMSCs on E-cadherin and α-SMA expression in human bronchial epithelial cells. A) Immunofluorescence staining; scale bars: 100 μm. B) Western blot analysis. *p<0.05 compared with control group; #p<0.05 compared model group.

Figure 5.

Effect of Hedgehog pathway inhibitor and UCMSCs on the expression of Shh, Gli1 and Snail in human bronchial epithelial cells. *p<0.05 compared with control group; ^#p<0.05 compared model group.