黄芩苷抑制人结肠黏膜上皮细胞的凋亡机制

Mechanism of baicalin inhibiting apoptosis of human colonic mucosal epithelial cells

[目的]探讨黄芩苷抑制人结肠黏膜上皮细胞凋亡的机制。[方法]黄芩苷处理人结肠黏膜上皮细胞,检测其凋亡水平与miRNA水平的变化。敲低筛选出的miRNA后,检测其凋亡水平与潜在靶标mRNA水平的变化。荧光素酶报告系统检测miRNA与潜在靶标的结合。[结果]黄芩苷处理人结肠黏膜上皮细胞后,人结肠黏膜上皮细胞的凋亡水平显著下降(0.35±0.11 vs 0.06±0.02,P<0.05)。黄芩苷处理人结肠黏膜上皮细胞后,miR-642a-3p、miR-484的表达水平显著下降(2.35±0.75 vs 0.51±0.18,1.94±0.63 vs 0.60±0.20,P<0.05)。敲低miR-484后,人结肠黏膜上皮细胞的凋亡水平显著降低(0.35±0.11 vs 0.07±0.02,P<0.05),PEA15和UAP1L1的水平显著升高(1.23±0.41 vs 2.45±0.64,0.65±0.23 vs 1.95±0.61,P<0.05)。敲低PEA15后,人结肠黏膜上皮细胞的凋亡水平显著升高(0.35±0.11 vs 0.67±0.020,P<0.05)。过表达PEA15后,人结肠黏膜上皮细胞的凋亡水平显著下降(0.35±0.10 vs 0.06±0.01,P<0.05)。miR-484靶向PEA15的3'端非编码区。[结论]黄芩苷处理人结肠黏膜上皮细胞后降低miR-484的表达、提高PEA15的表达,最终抑制了人结肠黏膜上皮细胞凋亡。

[Objective]To investigate the mechanism of baicalin inhibiting the apoptosis of human colon mucosal epithelial cells.[Method]Human colon mucosal epithelial cells were treated with baicalin to detect the changes of apoptosis and miRNA levels.After knockdown of the screened miRNAs,the changes of apoptosis level and the mRNA level of potential target were detected.The luciferase reporting system was used to detect whether miRNA directly binds to potential targets.[Result]The apoptotic level of human colon mucosal epithelial cells treated with baicalin was significantly decreased(0.35±0.11 vs 0.06±0.02,P<0.05).After baicalin treatment,the expression levels of miR-642A-3p and miR-484 were significantly decreased(2.35±0.75 vs 0.51±0.18,1.94±0.63 vs 0.60±0.20,P<0.05).After knockdown of miR-484,the apoptotic level of human colon mucosal epithelial cells was significantly decreased(0.35±0.11 vs 0.07±0.02,P<0.05).miR-484 knockdown significantly increased the levels of PEA15 and UAP1L1(1.23±0.41 vs 2.45±0.64,0.65±0.23 vs 1.95±0.61,P<0.05).The apoptosis level of human colon mucosal epithelial cells was significantly increased after PEA15 knockdown(0.35±0.11 vs 0.67±0.020,P<0.05).The apoptosis level of human colon mucosal epithelial cells was significantly decreased after overexpression of PEA15(0.35±0.10 vs 0.06±0.01,P<0.05).miR-484 targeted the 3'terminal non-coding region of PEA15.[Conclusion]Baicalin can decrease the expression of miR-484 and increase the expression of PEA15 in human colon mucosal epithelial cells,and finally inhibit the apoptosis of human colon mucosal epithelial cells.