摘要
目的探究芹菜素(AP)调节磷脂酰肌醇3-激酶/蛋白激酶B/雷帕霉素靶(PI3K/Akt/mTOR)信号通路对慢性湿疹(CE)大鼠炎症反应的影响。方法试验分为对照组(NC组)、慢性湿疹模型组(CE组)、芹菜素低剂量处理组(L-AP组)、芹菜素高剂量处理组(H-AP组)、芹菜素高剂量+Akt激活剂SC79处理组(H-AP+SC组),每组12只。对各组大鼠进行耳变应评分;计算免疫器官胸腺、脾脏指数;HE染色检测大鼠皮肤组织病理变化;ELISA试剂盒测定大鼠血清中炎性因子白细胞介素-1β(IL-1β)、IL-6、肿瘤坏死因子-α(TNF-α)水平;试剂盒测定大鼠血清中谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、丙二醛(MDA)水平;Western blot法检测大鼠皮肤组织匀浆中磷酸化PI3K(p-PI3K)、PI3K、磷酸化Akt(p-Akt)、Akt、磷酸化mTOR(p-mTOR)和mTOR蛋白水平。结果与NC组比较,CE组大鼠皮肤组织表皮角化不全,颗粒层增厚且有大量炎性细胞浸润,耳变应评分(1、7、14、28 d)、脾脏指数、胸腺指数、IL-1β、IL-6、TNF-α、MDA、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR水平升高,GSHPx、CAT、SOD水平降低,差异均有统计学意义(t=33.418、40.639、43.453、47.331、29.230、25.804、23.839、21.004、23.292、29.250、28.234、19.141、14.448、30.165、26.123、24.741,P均<0.05)。与CE组比较,L-AP组、H-AP组大鼠表皮角化程度改善,颗粒层变薄且炎性细胞减少,耳变应评分(7、14、28 d)、脾脏指数、胸腺指数、IL-1β、IL-6、TNF-α、MDA、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR水平降低,GSH-Px、CAT、SOD水平升高,差异均有统计学意义(t_(CE组vs.L-AP组)=8.154、12.554、17.703、28.676、22.530、32.485、12.846、23.458、10.976、20.990、12.758、20.081、7.067、16.987、9.304,t_(CE组vs.H-AP组)=19.706、13.812、22.883、11.728、23.022、9.390、16.974、6.192、13.416、12.189、27.622、13.885、23.308、9.213、17.344,P均<0.05),且H-AP组上述指标变化较L-AP组显著,差异均有统计学意义(P均<0.05)。与H-AP组比较,H-AP+SC组大鼠表皮角化程度加重、颗粒层增厚且炎性细胞增多,耳变应评分(7、14、28 d)、脾脏指数、胸腺指数、IL-1β、IL-6、TNF-α、MDA、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR水平升高,GSH-Px、CAT、SOD水平降低,差异均有统计学意义(P均<0.05)。结论AP可能通过抑制PI3K/Akt/mTOR信号通路,减轻CE大鼠炎症反应。
Objective To explore the influence of apigenin(AP)on inflammatory response in chronic eczema(CE)rats by regulating phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)signaling pathway.Methods The experiment was separated into control group(NC group),chronic eczema model group(CE group),apigenin low-dose treatment group(L-AP group),apigenin high-dose treatment group(H-AP group),and apigenin high-dose treatment+Akt activator SC79 treatment group(H-AP+SC group).Ear allergy scores were performed on the rats in each group;the thymus and spleen indexes of immune organs were calculated;HE staining was applied to detect the pathological changes of rat skin tissue;ELISA kits were applied to measure the levels of inflammatory factors interleukin-1β(IL-1β),IL-6 and tumor necrosis factor-α(TNF-α)in rat serum;the kit was applied to measure the levels of glutathione peroxidase(GSH-Px),catalase(CAT),superoxide dismutase(SOD)and malondialdehyde(MDA)in rat serum;Western-blot method was applied to detect the protein levels of phosphorylated PI3K(p-PI3K),PI3K,phosphorylated Akt(p-Akt),Akt,phosphorylated mTOR(p-mTOR)and mTOR in rat skin tissue homogenate.Results Compared with the NC group,the skin tissue of the CE group had parakeratosis,the granular layer was thickened and a large number of inflammatory cells were infiltrated;the ear allergy score(1,7,14,28 d),spleen index,thymus index,and the levels of IL-1β,IL-6,TNF-α,MDA,p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR elevated;the levels of GSH-Px,CAT and SOD decreased(t=33.418,40.639,43.453,47.331,29.230,25.804,23.839,21.004,23.292,29.250,28.234,19.141,14.448,30.165,26.123,24.741,all P<0.05);compared with the CE group,the degree of keratinization in the epidermis of rats in the L-AP group and the H-AP group were improved,the granular layer was thinner,and the inflammatory cells decreased,the ear allergy score(7,14,28 d),spleen index,thymus index,and the levels of IL-1β,IL-6,TNF-α,MDA,p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR reduced;the levels of GSH-Px,CAT and SOD increased(t=8.154,12.554,17.703,28.676,22.530,32.485,12.846,23.458,10.976,20.990,12.758,20.081,7.067,16.987,9.304,19.706,13.812,22.883,11.728,23.022,9.390,16.974,6.192,13.416,12.189,27.622,13.885,23.308,9.213,17.344;all P<0.05),and the changes of the above indicators in the H-AP group were obviously higher than those in the L-AP group(P<0.05);compared with the H-AP group,the degree of keratinization in the epidermis of rats in the H-AP+SC group were aggravated,the granular layer was thickened,and the inflammatory cells increased;the ear allergy score(7,14,28 d),spleen index,thymus index,and the levels of IL-1β,IL-6,TNF-α,MDA,p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR increased;the levels of GSH-Px,CAT and SOD decreased(all P<0.05).Conclusion AP might reduce the inflammatory response in CE rats by inhibiting the PI3K/Akt/mTOR signaling pathway.
作者
王杰
段妍
李睿亚
WANG Jie;DUAN Yan;LI Ruiya(Department of Dermatology,People's Hospital of Inner Mongolia Autonomous Region,Hohhot,Inner Mongolia 010020,China)
出处
《热带医学杂志》
CAS
2023年第10期1358-1363,1501,共7页
Journal of Tropical Medicine
基金
内蒙古自治区自然科学基金(2018LH08016)
作者简介
王杰(1984-),女,硕士,副主任医师,研究方向:皮肤病与性病学;通信作者:段妍,E-mail:136399711@sohu.com
