绿原酸通过AKT信号通路抑制高糖环境下人牙周膜细胞凋亡

Chlorogenic acid inhibits apoptosis of human periodontal ligament cells in high glucose environment via AKT signaling pathway

目的以体外培养的人牙周膜成纤维细胞(hPDLCs)为实验对象,探讨绿原酸对高糖环境下hPDLCs凋亡及蛋白激酶B(AKT)信号通路的影响。方法体外培养hPDLCs,分别用正常糖(5.5 mmol/L)、高糖(25.0 mmol/L)或高糖联合绿原酸(高糖,25.0 mg/ml;绿原酸,1 mg/ml)处理hPDLCs。细胞凋亡试验检测不同组hPDLCs凋亡情况,同时蛋白免疫印迹法检测各组泛AKT及磷酸化AKT(p-AKT)蛋白的表达情况。符合正态分布的计量资料以均数±标准差(xˉ±s)表示,组间比较采用独立样本t检验。结果与正常糖组相比,高糖可显著促进hPDLCs的凋亡,凋亡率从(6.66±0.18)%提高至(16.72±0.50)%,差异有统计学意义(t=32.789,P<0.001),同时p-AKT蛋白的表达水平显著降低。而与高糖组相比,绿原酸能抑制高糖环境下hPDLCs的凋亡,凋亡率从(16.72±0.50)%降低至(11.32±0.17)%,差异有统计学意义(t=17.710,P<0.001),同时回救高糖环境下下调的p-AKT蛋白的表达水平。结论绿原酸能抵抗高糖诱导的hPDLCs凋亡,可能是通过激活AKT信号通路实现的。

Objective To explore the effects of chlorogenic acid on the apoptosis of human periodontal ligament cells(hPDLCs)in high glucose environment and AKT signaling pathway,with in vitro hPDLCs as the experimental objects.Methods The hPDLCs were cultured in vitro and treated with normal glucose(5.5 mmol/L),high glucose(25.0 mmol/L),and high glucose and chlorogenic acid(high glucose,25.0 mmol/L;chlorogenic acid,1 mg/ml).The apoptosis assays of hPDLCs in different groups were conducted,and the expressions of p-AKT and AKT protein were detected by Western blot.The measurement data of normal distribution were expressed as(xˉ±s),and were compared between the groups by independent-sample t test.Results The apoptosis rate was(6.66±0.18)%in the normal glucose group,and was in the high glucose group,with a statistical difference(t=32.789,P<0.001),indicating that high glucose could significantly promote the apoptosis of hPDLCs.The expression level of p-AKT protein in the high glucose group was significantly lower than that in the normal glucose group.The apoptosis rate in the high glucose and chlorogenic acid group was lower than that in the high glucose group[(11.32±0.17)%],with a statistical difference(t=17.710,P<0.001),indicating that chlorogenic acid could inhibit hPDLCs'apoptosis caused by high glucose and up-regulate the expression of p-AKT protein in high glucose environment.Conclusion Chlorogenic acid can inhibit the apoptosis of hPDLCs,which may be achieved by activating the AKT signaling pathway.