Xie Zhiyin, Zhou Yu, Duan Xingwu, Yang Lirong
a Department of Dermatology, Beijing Changping Hospital of Traditional Chinese Medicine , Beijing , China.
b Department of Dermatology, Dongzhimen Hospital, Affiliated to Beijing University of Traditional Chinese Medicine , Beijing , China.
J Recept Signal Transduct Res. 2019 Apr;39(2):134-145. doi: 10.1080/10799893.2019.1638402. Epub 2019 Jul 29.
Epidermal cells play an important role in regulating the regeneration of skin after burns and wounds. The aim of our study is to explore the role of Tanshinone IIA (Tan IIA) in the apoptosis of epidermal HaCaT cells induced by HO, with a focus on mitochondrial homeostasis and inverted formin-2 (INF2). Cellular viability was determined using the MTT assay, TUNEL staining, western blot analysis and LDH release assay. Adenovirus-loaded INF2 was transfected into HaCaT cells to overexpress INF2 in the presence of Tan IIA treatment. Mitochondrial function was determined using JC-1 staining, mitochondrial ROS staining, immunofluorescence and western blotting. Oxidative stress promoted the death of HaCaT cells and this effect could be reversed by Tan IIA. At the molecular levels, Tan IIA treatment sustained mitochondrial energy metabolism, repressed mitochondrial ROS generation, stabilized mitochondrial potential, and blocked the mitochondrial apoptotic pathway. Furthermore, we demonstrated that Tan IIA modulated mitochondrial homeostasis via affecting INF2-related mitochondrial stress. Overexpression of INF2 could abolish the protective effects of Tan IIA on HaCaT cells viability and mitochondrial function. Besides, we also reported that Tan IIA regulated INF2 expression via the ERK pathway; inhibition of this pathway abrogated the beneficial effects of Tan IIA on HaCaT cells survival and mitochondrial homeostasis. Overall, our results indicated that oxidative stress-mediated HaCaT cells apoptosis could be reversed by Tan IIA treatment via reducing INF2-related mitochondrial stress in a manner dependent on the ERK signaling pathway.
表皮细胞在烧伤和创伤后皮肤再生的调节中发挥着重要作用。我们研究的目的是探讨丹参酮IIA（Tan IIA）在过氧化氢诱导的表皮HaCaT细胞凋亡中的作用，重点关注线粒体稳态和倒转肌动蛋白2（INF2）。使用MTT法、TUNEL染色、蛋白质免疫印迹分析和乳酸脱氢酶释放测定法测定细胞活力。在Tan IIA处理的情况下，将携带腺病毒的INF2转染到HaCaT细胞中以过表达INF2。使用JC-1染色、线粒体活性氧染色、免疫荧光和蛋白质免疫印迹法测定线粒体功能。氧化应激促进了HaCaT细胞的死亡，而Tan IIA可以逆转这种作用。在分子水平上，Tan IIA处理维持了线粒体能量代谢，抑制了线粒体活性氧的产生，稳定了线粒体膜电位，并阻断了线粒体凋亡途径。此外，我们证明Tan IIA通过影响与INF2相关的线粒体应激来调节线粒体稳态。INF2的过表达可以消除Tan IIA对HaCaT细胞活力和线粒体功能的保护作用。此外，我们还报道Tan IIA通过ERK途径调节INF2的表达；抑制该途径消除了Tan IIA对HaCaT细胞存活和线粒体稳态的有益作用。总体而言，我们的结果表明，Tan IIA处理可以通过依赖ERK信号通路的方式减少与INF2相关的线粒体应激，从而逆转氧化应激介导的HaCaT细胞凋亡。
