阿霉素促进心脏成纤维细胞分泌IL-1β和Ⅰ型胶原蛋白的机制研究  

A study on the mechanism of doxorubicin promoting IL-1β and collagen type Ⅰ secretion in cardiac fibroblasts

作　　者：罗远良 李梦思 滕嘉硕 戴晓萌 唐翔栩 邢云 吕秀秀 LUO Yuan-liang;LI Meng-si;TENG Jia-shuo;DAI Xiao-meng;TANG Xiang-xu;XING Yun;LV Xiu-xiu(Department of Pathophysiology,Key Laboratory of State Administration of Traditional Chinese Medicine of the People's Re-public of China,School of Medicine,Jinan University,Guangzhou 510632,China)

机构地区：[1]暨南大学基础医学院病理生理学系,国家中医药管理局病理生理重点实验室,广东广州510000

出　　处：《中国病理生理杂志》2020年第9期1543-1550,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81300118)。

摘　　要：目的:观察阿霉素/多柔比星(DOX)对心脏成纤维细胞产生炎症细胞因子和胶原蛋白的影响及其机制。方法:体外培养SD大鼠乳鼠心脏成纤维细胞,免疫荧光染色标记波形蛋白鉴定细胞;CCK-8法检测DOX对心脏成纤维细胞的毒性;annexin V-FITC/PI双染后流式细胞术检测细胞凋亡;ELISA法检测细胞上清中炎症因子含量;免疫荧光标记法检测心脏成纤维细胞肌化和线粒体活性氧簇(mROS)释放的情况;Western blot法检测核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体活化相关蛋白的水平。结果:(1)与对照组比较,DOX显著抑制心脏成纤维细胞增殖(P<0.05),但对细胞凋亡并没有显著影响(P>0.05);(2)DOX促进心脏成纤维细胞释放白细胞介素1β(IL-1β)和IL-6(P<0.05);(3)DOX促进心脏成纤维细胞肌化,与对照组比较,DOX处理组细胞α-平滑肌肌动蛋白、Ⅰ型胶原蛋白和转化生长因子β表达明显增加(P<0.05);(4)与对照组比较,心脏成纤维细胞在DOX的作用下,mROS、NLRP3及cleaved caspase-1水平均明显增加(P<0.05)。结论:DOX通过促进mROS释放及NLRP3炎症小体活化而促进心脏成纤维细胞分泌IL-1β和Ⅰ型胶原蛋白。AIM:To observe the effect of adriamycin/doxorubicin(DOX)on the production of inflammatory cytokines and collagen in cardiac fibroblasts and its mechanism.METHODS:Neonatal SD rat cardiac fibroblasts were isolated,cultured,and identified by immunofluorescence staining with monoclonal antibodies against vimentin observed under a confocal laser-scanning microscope.The Cell Counting Kit-8 assay was used to detect the toxicity of DOX on cardiac fibroblasts,and flow cytometry with annexin V-FITC/PI double staining was used to detect apoptosis.ELISA was used to detect the release of inflammatory factors in the supernatant of cultured cells.Immunofluorescence labeling assay was used to detected α-smooth muscle actin(α-SMA)expression and mitochondrial reactive oxygen species(mROS)in the cells.Western blot was used to detect the expression of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome-related proteins in cardiac fibroblasts.RESULTS:(1)Compared with the control group,DOX inhibited the proliferation of cardiac fibroblasts(P<0.05),but had no significant effect on apoptosis(P>0.05).(2)Treatment with DOX promotes the release of proinflammatory factors interleukin-1β(IL-1β)and IL-6 in cardiac fibroblasts(P<0.05).(3)The expression ofα-SMA,collagen type Ⅰ and transforming growth factor-βin DOX treatment group increased significantly compared with control group(P<0.05).(4)Compared with the control group,the levels of mROS,cellular NLRP3 and cleaved caspase-1 in cardiac fibroblasts increased significantly after DOX treatment.CONCLUSION:Doxorubicin promotes cardiac fibroblasts to secrete IL-1β and collagen type Ⅰ by promoting mROS production and activating NLRP3 inflammasome.

关 键 词：阿霉素/多柔比星 心脏成纤维细胞 非细菌性炎症 纤维化 

分 类 号：R965[医药卫生—药理学] R363[医药卫生—药学]