基于AMPK/mTOR/NLRP3信号通路探究复方黄柏液对痤疮丙酸杆菌诱导的皮肤炎症的影响  

Effects of compound Cortex Phellodendri liquid on Propionibacterium acnes-induced skin inflammation based on AMPK/mTOR/NLRP3 signaling pathway

作　　者：李耕[1] 蒋晓蕾[2] 禹红[1] LI Geng;JIANG Xiaolei;YU Hong(Department of Laboratory,Wuhan Hospital of Traditional Chinese Medicine,Wuhan 430010,China;Department of Dermatology,Wuhan Hospital of Traditional Chinese Medicine,Wuhan 430010,China)

机构地区：[1]武汉市中医医院检验科,430010 [2]武汉市中医医院皮肤科,430010

出　　处：《免疫学杂志》2022年第10期829-837,共9页Immunological Journal

基　　金：武汉市医学科研项目(WZ20C39)。

摘　　要：目的基于腺苷酸活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)/NOD样受体蛋白3(NLRP3)通路探究复方黄柏液(CCPF)对痤疮丙酸杆菌诱导的皮肤炎症的影响。方法大鼠分为NC组、Model组、CCPF组(5 ml)、CCPF+compound C(AMPK抑制剂)组(5 ml CCPF+0.25 mg/kg compound C)、CCPF+MHY1485(m TOR激活剂)组(5 ml CCPF+10 mg/kg MHY1485),每组12只。除NC组外,其它组大鼠均需向右耳皮内注射痤疮丙酸杆菌混悬液以构建痤疮模型。建模成功后,进行给药处理,每天1次,持续14 d。观察各组大鼠耳部炎症、红肿情况;利用游标卡尺测量大鼠右耳耳廓的厚度,并计算耳廓肿胀率、耳廓痤疮数;HE染色检测耳廓组织病理变化并计数炎性细胞数;ELISA法检测大鼠血清中睾酮(T)、雌二醇(E2)及肿瘤坏死因子-α(TNF-α)、白介素-8(IL-8)水平;免疫组化检测大鼠耳组织中TNF-α、IL-8蛋白水平;Western blot检测p-AMPK、AMPK、p-mTOR、mTOR、NLRP3、半胱氨酸天冬氨酸蛋白酶1(caspase-1)蛋白表达。结果与NC组比较,Model组大鼠耳廓组织红肿明显,注射部位有脓疱产生,血管充血及炎性细胞浸润明显,耳廓厚度、耳廓肿胀率、耳廓痤疮数、T、TNF-α、IL-8水平、p-mTOR/mTOR、NLRP3、caspase-1蛋白表达升高,E2、p-AMPK/AMPK水平降低(P<0.05);与Model组比较,CCPF组的上述指标均得到改善(P<0.05);compound C或MHY1485逆转了CCPF对痤疮丙酸杆菌诱导的大鼠皮肤炎症的改善作用。结论CCPF可能通过激活AMPK/mTOR通路来抑制NLRP3炎症小体的激活,从而改善痤疮丙酸杆菌诱导的皮肤炎症。This study was performed to explore the impact of compound Cortex Phellodendri liquid(CCPF)on Propionibacterium acnes(P.acnes)-induced skin inflammation based on AMP-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)/NOD-like receptor protein 3(NLRP3)pathway.Rats were recruited and divided into NC group,model group,CCPF group,CCPF+compound C(AMPK inhibitor compound C)group,CCPF+MHY1485(mTOR activator MHY1485)group,with 12 mice in each group.Except for the NC group,the other groups were injected with the P.acnes suspension intradermally into the right ear to construct an acne model.After successful modeling,drug treatment was carried out once a day for 14 days.The ear inflammation,redness and swelling of the rats in each group were observed;the thickness of the right ear auricle was measured with a vernier caliper,and the swelling rate of the auricle and theauricle acne count were calculated;the pathologicalchanges of the auricle was measured by HE staining andthe inflammatory cells were counted;the levels of serum testosterone(T)and estradiol(E2),TNF-αand IL-8 were measured by ELISA;immunohistochemistry was used todetect TNF-αand IL-8 protein levels in rat ear tissue;the protein expressions of p-AMPK,AMPK,p-mTOR,mTOR,NLRP3 and caspase-1 were measured by Western blotting.Compared with the NC group,the auricle tissue of model group was obviously red and swollen,pustules were formed at the injection site,vascular congestion and inflammatory cell infiltration were obvious,the thickness of the auricle,the swelling rate of the auricle,auricle acne count,the levels of T,TNF-α,IL-8,and the expression of p-mTOR/mTOR,NLRP3,caspase-1 proteins were increased,while the levels of E2 and p-AMPK/AMPK were decreased(P<0.05).As compared with the model group,these indexes mentioned above in CCPF group were all ameliorated(P<0.05).Compound C or MHY1485 could reverse the ameliorative effects of CCPF on P.acnes-induced skin inflammation in rats.Taken together,CCPF may activate the AMPK/mTOR pathway to inhibit the ac

关 键 词：复方黄柏液 腺苷酸活化蛋白激酶/哺乳动物雷帕霉素靶蛋白/NOD样受体蛋白3通路 痤疮丙酸杆菌 皮肤炎症 

分 类 号：R285.5[医药卫生—中药学]