目的 分析咖啡酸對UVB損傷人角質(zhì)形成細(xì)胞HaCaT的保護(hù)作用機(jī)制。方法 采用UVB燈照射建立HaCaT細(xì)胞UVB損傷模型。HaCaT細(xì)胞中分別加入濃度梯度為0（對照組）、5、10、20、40、80、160µ；mol/L的咖啡酸，計(jì)算細(xì)胞存活率，選擇最佳濃度進(jìn)行后續(xù)試驗(yàn)。將處于對數(shù)生長期HaCaT分為對照組、模型組、咖啡酸（10µ；mol/L）組，進(jìn)行蘇木精–伊紅（HE）染色，顯微鏡觀察細(xì)胞形態(tài)。按照試劑盒說明書操作，采用樣本蛋白濃度計(jì)算方法，測定細(xì)胞中過氧化氫酶（CAT）、超氧化物歧化酶（SOD）的含量。蛋白免疫印記檢測促分裂原活化蛋白激酶（MAPK）通路相關(guān)蛋白的表達(dá)。結(jié)果 咖啡酸10µ；mol/L組能夠減輕UVB對HaCaT細(xì)胞造成的損傷，修復(fù)HaCaT的細(xì)胞形態(tài)，增加抗凋亡蛋白Bcl-2在HaCaT細(xì)胞中的表達(dá)，減少細(xì)胞的凋亡?？Х人?0µ；mol/L組能夠顯著升高HaCaT中SOD、CAT含量，增強(qiáng)細(xì)胞中的抗氧化能力，降低MAPK亞族p38信號通路p-p38和p53蛋白在細(xì)胞中的表達(dá)。結(jié)論 咖啡酸可以抑制UVB對細(xì)胞的損傷，修復(fù)細(xì)胞的形態(tài)結(jié)構(gòu)，減少細(xì)胞的凋亡和增強(qiáng)細(xì)胞的抗氧化性，且可能通過MAPK信號通路的調(diào)控來減弱UVB對HaCaT細(xì)胞的損傷，而對UVB損傷后的HaCaT細(xì)胞具有保護(hù)作用。

Objective To analyze the protective mechanism of caffeic acid on HaCaT cells after UVB injury. Method UVB injury model of HaCaT cells was established by irradiation with UVB lamp. Caffeic acid was added to HaCaT cells with concentration gradients of 0 (control group), 5, 10, 20, 40, 80, and 160 µmol/L, respectively, to calculate the cell survival rate and select the final concentration for subsequent tests. HaCaT cells in logarithmic growth phase were divided into control group, model group, and caffeic acid 10 µmol/L group. Hematoxylin and eosin (HE) staining was performed to observe the cell morphology under microscope. Results The 10 µmol/L caffeic acid group could repair the damage caused by UVB to HaCaT cells, repair the cell morphology of HaCaT cells, increase the expression of anti-apoptotic protein Bcl-2 in HaCaT cells, and reduce cell apoptosis. Caffeic acid 10 µmol/L group can significantly increase the contents of SOD and CAT in HaCaT cells, enhance the antioxidant capacity of cells, and reduce the expression of p-P38 and P53 proteins in MAPK subgroup P38 signaling pathway in cells. Conclusion Caffeic acid can inhibit the damage of UVB to cells, repair the morphological structure of cells, reduce cell apoptosis and enhance the antioxidant capacity of cells. It may weaken the damage of UVB to HaCaT cells through the regulation of MAPK signal pathway, and have a protective effect on HaCaT cells damaged by UVB.

R966

云南中煙工業(yè)有限責(zé)任公司科技項(xiàng)目（2020CP02.2020539200340205）