桃红四物汤通过调控自噬-NLRP3 炎性小体通路对氧葡萄糖剥夺/复氧损伤 PC12 细胞的保护作用。
Protection of Taohong Siwu Decoction on PC12 cells injured by oxygen glucose deprivation/reperfusion via mitophagy-NLRP3 inflammasome pathway in vitro.
机构信息
College of Pharmacy, Anhui University of Chinese Medicine, China.
College of Pharmacy, Anhui University of Chinese Medicine, China; MOE-Anhui Joint Collaborative Innovation Center for Quality Improvement of Anhui Genuine Chinese Medicinal Materials, China; Anhui Province Key Laboratory of Chinese Medicinal Formula, China; Institute of Traditional Chinese Medicine Resources Protection and Development, China; Anhui Province Key Laboratory of Traditional Chinese Medicine Decoction Pieces of New Manufacturing Technology, China.
出版信息
J Ethnopharmacol. 2023 Jan 30;301:115784. doi: 10.1016/j.jep.2022.115784. Epub 2022 Oct 4.
ETHNOPHARMACOLOGICAL RELEVANCE
Taohong Siwu Decoction (THSWD) is a traditional Chinese medicine formula used to invigorate blood circulation and resolve blood stasis. It consists of Paeonia lactiflora Pall., Conioselinum anthriscoides (H.Boissieu) Pimenov & Kljuykov, Rehmannia glutinosa (Gaertn.) DC., Prunus persica (L.) Batsch, Angelica sinensis (Oliv.) Diels, and Carthamus creticus L. in the ratio of 3:2:4:3:3:2. THSWD is a common prescription for the treatment of ischemic stroke.
AIM OF THE STUDY
To study the protective effect and mechanism of Taohong Siwu Decoction (THSWD) on PC12 cells damaged by oxygen glucose deprivation/reperfusion (OGD/R).
MATERIALS AND METHODS
OGD/R model of PC12 cells was used to simulate ischemia-reperfusion (I/R) injury of nerve cells in vitro. The experiment was grouped as follows: control, OGD/R and OGD/R + THSWD (5%, 10% and 15%) group. Oxygen and glucose was restored for 24 h after 4-6 h of deprivation. The severity of damage to PC12 cells was evaluated by CCK8, flow cytometry and lactate dehydrogenase (LDH). Mitochondrial morphology and function were examined by transmission electron microscopy (TEM), ATP and mitochondrial membrane potential (MMP) assay kits. Cellular autophagy and NLRP3 inflammasome-associated proteins were detected by Western blot and immunofluorescence staining.
RESULTS
THSWD treatment improved the survival rate of PC12 cells injured by OGD/R, reduced cell damage and apoptosis. Moreover, ATP, MMP and the expression of autophagy marker proteins (LC3-II/LC3-I, Beclin1, Atg5) and mitophagy marker proteins (Parkin and PINK-1) was significantly elevated. The reactive oxygen species (ROS), NLRP3 inflammasome and pro-inflammatory cytokines induced by OGD/R were distinctly reduced. In contrast, these above beneficial effects of THSWD on mitochondrial autophagy and NLRP3 inflammasome were reversed by mitochondrial division inhibitory factor 1 (Mdivi-1).
CONCLUSION
THSWD protects PC12 cells against OGD/R injury by heightening mitophagy and suppressing the activation of NLRP3 inflammasome.
ETHNOPHARMACOLOGICAL RELEVANCE
桃红四物汤（THSWD）是一种传统的中药方剂，用于活血化瘀。它由白芍、苍术、熟地黄、桃仁、当归和红花按 3:2:4:3:3:2 的比例组成。THSWD 是治疗缺血性中风的常用方剂。
AIM OF THE STUDY
研究桃红四物汤（THSWD）对氧葡萄糖剥夺/再灌注（OGD/R）损伤的 PC12 细胞的保护作用及其机制。
MATERIALS AND METHODS
采用氧葡萄糖剥夺/再灌注（OGD/R）模型模拟体外神经细胞缺血再灌注（I/R）损伤。实验分组如下：对照组、OGD/R 组和 OGD/R+THSWD（5%、10%和 15%）组。缺氧 4-6h 后恢复氧和葡萄糖 24h。通过 CCK8、流式细胞术和乳酸脱氢酶（LDH）评估 PC12 细胞损伤程度。通过透射电镜（TEM）、ATP 和线粒体膜电位（MMP）检测试剂盒检测线粒体形态和功能。通过 Western blot 和免疫荧光染色检测细胞自噬和 NLRP3 炎性体相关蛋白。
RESULTS
THSWD 治疗可提高 OGD/R 损伤的 PC12 细胞存活率，减轻细胞损伤和凋亡。此外，ATP、MMP 和自噬标志物蛋白（LC3-II/LC3-I、Beclin1、Atg5）和线粒体自噬标志物蛋白（Parkin 和 PINK-1）的表达明显升高。OGD/R 诱导的活性氧（ROS）、NLRP3 炎性体和促炎细胞因子明显减少。相反，THSWD 对线粒体自噬和 NLRP3 炎性体的上述有益作用被线粒体分裂抑制因子 1（Mdivi-1）逆转。
CONCLUSION
THSWD 通过增强线粒体自噬和抑制 NLRP3 炎性体的激活来保护 PC12 细胞免受 OGD/R 损伤。
Zotero 插件