目的 探討柚皮苷通過P38 MAPK/NF-κB通路對脂多糖（LPS）致HaCaT細(xì)胞炎癥損傷的抑制作用。方法 LPS（0、0.1、1.0、10.0、20.0 μg/mL）孵育24 h刺激人永生化角質(zhì)細(xì)胞HaCaT，模擬銀屑病模型，MTT法觀察細(xì)胞存活率變化，Western bloting法檢測白介素（IL）-6和NF-κB蛋白表達(dá)；MTT法觀察柚皮苷（0、5、10、20、40、80、160、320μmol/L）作用24 h對HaCaT存活率的影響；選擇LPS、柚皮苷最佳作用濃度。柚皮苷（20和40 μmol/L）作用銀屑病模型HaCaT細(xì)胞24 h，實(shí)時(shí)熒光定量PCR（qRT-PCR）法檢測IL-6、IL-1β、IL-17和腫瘤壞死因子-α（TNF-α）的mRNA表達(dá)水平變化；Western blotting法檢測細(xì)胞核內(nèi)轉(zhuǎn)錄因子NF-κB p65、P38 MAPK磷酸化蛋白水平以及炎癥因子IL-6蛋白水平。結(jié)果 LPS增加HaCaT細(xì)胞存活率，并上調(diào)NF-κB p65和IL-6蛋白表達(dá)，呈劑量相關(guān)性，炎癥反應(yīng)在20 μg/mL達(dá)到高峰；5~160 μmol/L柚皮苷對HaCaT細(xì)胞無明顯毒性作用。與模型組比較，柚皮苷能夠顯著抑制炎癥因子IL-6、IL-1β、IL-17和TNF-α的轉(zhuǎn)錄水平（P<0.05）；同時(shí)能夠顯著抑制LPS誘導(dǎo)的NF-κB p65和P38 MAPK磷酸化蛋白水平、抑制IL-6蛋白的表達(dá)（P<0.05）。結(jié)論 柚皮苷抑制脂多糖致HaCaT細(xì)胞炎癥反應(yīng)，發(fā)揮改善銀屑病的作用，機(jī)制可能與抑制P38 MAPK/NF-κB信號通路有關(guān)。

Objective To investigate the inhibitory effects of naringin on HaCaT cells inflammation induced by lipopolysaccharide (LPS) through NF-κB/P38 pathway. Methods LPS (0, 0.1, 1.0, 10.0, 20.0 μg/mL) incubated for 24 hours stimulated human immortalized keratinocytes HaCaT to simulate psoriasis model. MTT method was used to observe the changes of cell survival rate, and Western blotting method was used to detect the expression of IL-6 and NF-κB protein. MTT method was used to observe the effect of naringin (0, 5, 10, 20, 40, 80, 160, 320 μmol/L) on the survival rate of HaCaT cells. The optimal concentration of LPS and naringin was selected. Naringin (20 and 40 μmol/L) acted on HaCaT cells of psoriasis model for 24 hours. Reverse transcription polymerase chain reaction (qRT-PCR) was performed to determine the mRNA expression levels of IL-6, IL-1β, IL-17 and TNF-α. Western blotting were used to detect the phosphorylation levels of P38 MAPK and NF-κB p65 and the inflammatory factor IL-6. Results LPS increased the survival rate of HaCaT cells, and up-regulated the protein expression of NF-κB p65 and IL-6, which was dose-related, and the inflammatory response peaked at 20 μg/mL. 5~160 μmol/L naringin had no obvious toxic effect on HaCaT cells. Compared with model group, naringin can significantly inhibit the transcription of inflammatory cytokines IL-6, IL-1β, IL-17 and TNF-α (P<0.05). Meanwhile, naringin can significantly inhibit LPS-induced phosphorylation of P38 MAPK and NF-κB p65, inhibiting the expression of IL-6 protein (P<0.05). Conclusion Naringin inhibits inflammation of HaCaT cells induced by lipopolysaccharide and plays a role in improving psoriasis. The mechanism may be related to the inhibition of P38 MAPK/NF-κB signaling pathway.

R962.2

黑龍江省屬高等學(xué)?；究蒲袠I(yè)務(wù)費(fèi)科研項(xiàng)目（2015-KYYWF-002）；牡丹江醫(yī)學(xué)院研究生創(chuàng)新科研項(xiàng)目（2017YJSCX-20MY）