目的 探讨芍药苷通过调节成纤维细胞（HSF）来源外泌体（Exo）微小RNA-181a-5p（miR-181a-5p）对中波紫外线（UVB）诱导的角质形成细胞HaCaT光老化的影响。方法 以HaCaT细胞为研究对象，将其分为对照组、模型组、芍药苷组、HSF-Exo组、芍药苷＋HSF-Exo组、过表达阴性对照（OE-NC）组、OE-miR-181a-5p组。qRT-PCR法检测miR-181a-5p mRNA表达；MTT检测HaCaT细胞增殖情况；β-半乳糖苷酶（SA-β-Gal）染色法检测HaCaT细胞衰老情况；流式细胞仪检测HaCaT细胞凋亡率；DCFH-DA探针检测HaCaT细胞中活性氧（ROS）水平；ELISA检测HaCaT细胞中超氧化物歧化酶（SOD）、丙二醛（MDA）、白细胞介素（IL）-6、IL-1β、肿瘤坏死因子-α（TNF-α）表达；Western blotting检测HaCaT细胞中p16、p21、I型胶原蛋白α1链基因（COL1A1）、基质金属蛋白酶-1（MMP-1）蛋白表达情况。结果 与模型组比较，芍药苷组、HSF-Exo组、芍药苷＋HSF-Exo组增殖率、SOD、COL1A1含量显著升高，衰老细胞比例、凋亡率、ROS荧光强度，MDA、IL-6、IL-1β、TNF-α含量，p16、p21、MMP-1蛋白相对表达量，miR-181a-5p表达量显著降低（P＜0.05）；与芍药苷组、HSF-Exo组比较，芍药苷＋HSF-Exo组增殖率、SOD、COL1A1含量显著升高，衰老细胞比例、凋亡率、ROS荧光强度，MDA、IL-6、IL-1β、TNF-α含量，p16、p21、MMP-1蛋白相对表达量，miR-181a-5p表达量显著降低（P＜0.05）；与芍药苷＋HSF-Exo组、OE-NC组比较，OE-miR-181a-5p组增殖率、SOD、COL1A1含量显著降低，衰老细胞比例、凋亡率、ROS荧光强度，MDA、IL-6、IL-1β、TNF-α含量，p16、p21、MMP-1蛋白相对表达量，miR-181a-5p表达量显著升高（P＜0.05）。结论 芍药苷通过调节HSF细胞来源Exo miR-181a-5p抑制UVB诱导的角质形成细胞光老化。

Objective To investigate the effect of paeoniflorin on the photoaging of keratinocytes induced by UVB through regulating miR-181a-5p in exosome (Exo) derived from human skin fibroblasts (HSF). Methods Using HaCaT cells as the research object, diveded into the control group, model group, paeoniflorin group, HSF-Exo group, paeoniflorin + HSF-Exo group, OE-NC group, and OE-miR-181a-5p group. The expression of miR-181a-5p mRNA was detected by qRT-PCR. The proliferation of HaCaT cells was examined by MTT assay. SA-β-Gal staining method was implemented to detect the senescence of HaCaT cells. Flow cytometer was used to detect the apoptosis of HaCaT cells. DCFH-DA probe was used to detect the level of ROS in HaCaT cells. ELISA was employed to measure the content of SOD, MDA, IL-6, IL-1β and TNF-α in HaCaT cells. Western blotting was implemented to detect the protein expressions of p16, p21, COL1A1 and MMP-1 in HaCaT cells. Results Compared with model group, growth rate, content of SOD and COL1A1 in the paeoniflorin group, HSF-Exo group, paeoniflorin + HSF-Exo group were increased, while the proportion of senescent cells, apoptosis rate, ROS fluorescence intensity, content of MDA, IL-6, IL-1β, TNF-α, the protein expression of p16, p21, MMP-1, and miR-181a-5p expression were decreased (P < 0.05). Compared with paeoniflorin group and HSF-Exo group, growth rate, content of SOD and COL1A1 in the paeoniflorin + HSF-Exo group were increased, while the proportion of senescent cells, apoptosis rate, ROS fluorescence intensity, content of MDA, IL-6, IL-1β, TNF-α, the protein expression of p16, p21, MMP-1, and miR-181a-5p expression were decreased (P < 0.05). Compared with paeoniflorin + HSF-Exo group and OE-NC group, growth rate, content of SOD and COL1A1 in the OE-miR-181a-5p group were decreased, while the proportion of senescent cells, apoptosis rate, ROS fluorescence intensity, content of MDA, IL-6, IL-1β, TNF-α, the protein expression of p16, p21, MMP-1, and miR-181a-5p expression were increased (P < 0.05). Conclusion Paeoniflorin inhibits UVB-induced photoaging of keratinocytes by regulating HSF-derived Exo miR-181a-5p.

武汉市医学科研项目立项（WX20Q03）