骨髓来源的间充质干细胞对变应性鼻炎大鼠鼻黏膜上皮细胞的调控作用

Regulatory effect of bone marrow-derived mesenchymal stem cells on nasal mucosal epithelial cells in rats with allergic rhinitis

目的研究骨髓间充质干细胞(MSCs)对变应性鼻炎(AR)大鼠鼻黏膜上皮细胞形态、增殖和分泌蛋白的影响。方法分离正常大鼠骨髓MSCs、正常大鼠鼻黏膜上皮细胞以及AR大鼠鼻黏膜上皮细胞并分别培养,细胞培养传代至符合要求时,将骨髓MSCs和AR大鼠鼻黏膜上皮细胞共培养,培养3 d、7 d、14 d时显微镜观察鼻黏膜上皮细胞形态,CCK-8法检测鼻黏膜上皮细胞增殖情况[设模型组(AR大鼠鼻黏膜上皮细胞)和模型MSCs组(AR大鼠鼻黏膜上皮细胞和骨髓MSCs共培养),以OD值表示],Western blot法检测鼻黏膜上皮细胞黏液分泌标志物黏蛋白5AC(MUC5AC)蛋白表达情况[设正常组(正常大鼠鼻黏膜上皮细胞)、正常MSCs组(正常大鼠鼻黏膜上皮细胞和骨髓MSCs共培养)、模型组(AR大鼠鼻黏膜上皮细胞)、模型MSCs组(AR大鼠鼻黏膜上皮细胞和骨髓MSCs共培养)]。结果骨髓MSCs与AR大鼠的鼻黏膜上皮细胞共培养14 d时,鼻黏膜上皮细胞明显增多,细胞生长旺盛,活性好,细胞形成大的集落群。培养14 d时,模型MSCs组的鼻黏膜上皮细胞增殖OD值为0.97±0.06,模型组为0.81±0.05,2组比较差异有统计学意义(P<0.05)。正常组和正常MSCs组鼻黏膜上皮细胞中MUC5AC蛋白相对表达量比较差异无统计学意义(P>0.05);模型MSCs组鼻黏膜上皮细胞中MUC5AC蛋白相对表达量明显低于模型组(P<0.05)。结论骨髓MSCs能促进AR大鼠鼻黏膜上皮细胞增殖,下调AR大鼠鼻黏膜上皮细胞中MUC5AC蛋白表达。

Objective It is to investigate the effects of bone marrow-derived mesenchymal stem cells(MSCs)on the morphology,proliferation and secretory proteins of nasal mucosal epithelial cells in rats with allergic rhinitis(AR).Methods MSCs in normal rats,nasal mucosal epithelial cells in normal rats and nasal mucosal epithelial cells in AR rats were separated and cultured respectively.When the cell culture and subculture met the requirements,the MSCs and AR rat nasal mucosal epithelial cells were co-cultured,and the morphological changes of the nasal mucosal epithelial cells were observed by microscope after 3,7 and 14 days of culture.The proliferation of nasal mucosal epithelial cells was detected by CCK8 method[A model group(AR rat nasal mucosal epithelial cells)and a model MSCs group(co-culture of AR rat nasal mucosal epithelial cells and bone marrow MSCs)were set up,expressed as OD values],the protein expression of mucin 5AC(MUC5AC),a marker of mucus secretion in nasal mucosal epithelial cells,was detected by Western blot assay[normal group(normal rat nasal mucosal epithelial cells),normal MSCs group(co-culture of normal rat nasal mucosal epithelial cells and bone marrow MSCs),model group(AR rat nasal mucosal epithelial cells),model MSCs group(co-culture of AR rat nasal mucosal epithelial cells and bone marrow MSCs)].Results When MSCs were co-cultured with nasal mucosal epithelial cells of AR rats for 14 days,the nasal mucosal epithelial cells were increased significantly,they formed a large colony with vigorous growth and viability.After 14 days of culture,the OD value of nasal mucosal epithelial cell proliferation was 0.97±0.06 in the model MSCs group and 0.81±0.05 in the model group,and the difference between the two groups was statistically significant(P<0.05).There was no statistically significant difference in the relative expression of MUC5AC protein in the nasal mucosal epithelial cells between the normal group and normal MSCs group(P>0.05);the relative expression of MUC5AC protein in the nasal mucosal epithelial cells of the model MSCs group was significantly lower than that of the model group(P<0.05).Conclusion MSCs can promote the proliferation of nasal mucosal epithelial cells and down-regulate the expression of MUC5AC in nasal mucosal epithelial cells in AR rats.