NLRP炎症小体在光老化皮肤成纤维细胞的表达及作用机制探讨  

Expression changes and mechanisms of NLRP inflammasomes in photoaged skin

作　　者：刘玉芳 赖维[1] 张杰 林瑶 谢小元[1] 郑跃[1] Liu Yufang;Lai Wei;Zhang Jie;Lin Yao;Xie Xiaoyuan;Zheng Yue(Department of Dermatology,the Third Affiliated Hospital,Sun Yat-sen University,Guangzhou 510630,China)

机构地区：[1]中山大学附属第三医院皮肤科,广州510630

出　　处：《中华医学美学美容杂志》2020年第6期534-537,共4页Chinese Journal of Medical Aesthetics and Cosmetology

基　　金：国家自然科学基金(81673085)。

摘　　要：目的探讨NLRP在光老化皮肤成纤维细胞表达变化和皮肤光老化机制中的作用。方法2018年8月至2019年4月,在中山大学附属第三医院泌尿外科取3例儿童包皮环切术后包皮组织,分离培养成纤维细胞,并分为UVA照射组和空白对照组,UVA照射组接受连续UVA照射诱导慢性光损伤,空白对照组不接受UVA照射,通过CCK8试剂盒、β-半乳糖苷酶染色(SA-β-Gal)及细胞凋亡率检测验证建模。Western免疫印迹试验检测NLRP1、2、3、8蛋白表达。高通量测序技术检测调控NLRP炎症小体的非编码RNA表达变化。结果UVA照射组细胞活性(70.0±4.7)%明显低于空白对照组(93.0±2.2)%,t=6.93,P<0.05;细胞老化率(83.1±4.4)%高于空白对照组老化率(9.2±0.85)%(t=25.21,P<0.05);细胞凋亡率(34.5±7.1)%高于空白对照组(9.3±2.2)%(t=6.42,P<0.05)。蛋白定量检测结果显示,UVA照射组NLRP1、2、3、8表达较空白对照组(0.75±0.43)均上调,分别为5.59±0.99、3.84±0.69、9.98±1.72、1.57±0.56。高通量测序技术检测UVA照射组与空白对照组之间差异表达LncRNA,发现可调控NLRP炎症小体的长链非编码RNA中差异表达LncRNA共23个,其中变化最明显的为lnc-NLRP1、lnc-NLRP3。结论反复UVA照射可上调NLRP炎症小体及其长链非编码LncRNA的表达,可通过参与活性氧自由基聚集、信号通路激活、自噬、基质金属蛋白酶/溶酶体组织蛋白酶家族表达及活性改变等机制,引起皮肤光老化。Objective To investigate the expression of NLRP inflammasome in skin photoaging mechanisms.Methods Some cultured human dermal fibroblasts were subjected to repetitive ultraviolet A(UVA)radiation(UVA radiation group)to establish a photoaging cell model,which was then evaluated by cell counting kit 8(CCK-8)assay,β-galactosidase staining and flow cytometry detection of apoptosis rate.Expression of NLRP1,NLRP2,NLRP3 and NLRP8 was detected via Western blot.High throughput sequencing was used to detect the expression of non-coding RNA(LncRNA)which regulates the inflammatory bodies of NLRP(Lnc-NLRP).The mean of two independent samples was compared by independent sample t test,and statistical significance was identified as P<0.05.Results Compared with that in the control group,the UVA radiation group showed significantly decreased cellular proliferative activity(70±4.7)%vs.(93±2.2)%,t=6.93,P<0.05),but significantly increased apoptotic rate(34.5±7.1)%vs.(9.3±2.2)%,t=6.42,P<0.05).and percentage ofβ-galactosidase(83.1±4.4)%vs.(9.2±0.85)%,t=25.21,P<0.05).NLRP1,NLRP2,NLRP3 and NLRP8 expression was increased to 5.59±0.99,3.84±0.69,9.98±1.72,1.57±0.56 compared to the control(0.75±0.43).Differentially expressed LncRNAs was detected via high throughput sequencing technique and found that 23 differentially expressed LncRNAs contributing to regulate NLRP inflammations among which the most obvious changes were lnc-NLRP1 and lnc-NLRP3.Conclusions Repeated UVA irradiation could up-regulate the expression of NLRP inflammasomes and LncNLRP which might take part in ROS aggregation,cell autophagy,MMPs/cathepsins expression and activity changes and signaling pathway activation process in skin photoaging mechanisms.

关 键 词：成纤维细胞 紫外线 NLRP炎症小体 光老化 机制 

分 类 号：R758.1[医药卫生—皮肤病学与性病学]