背景:红花弹力。(MD),She族使用的一种传统中药,已被用于治疗脑缺血再灌注(CIR)损伤,由于其在促进血液循环和消除血液淤积的功效;然而,MD治疗CIR损伤的疗效和机制尚不清楚。
目的:探讨MD对CI-R损伤的保护作用,除了对氧化应激的影响,内质网(ER)应激,和细胞凋亡。
方法:使用细胞实验和动物实验进行研究。CCK-8方法,免疫荧光染色,和流式细胞术分析含药血清对氧-葡萄糖剥夺/再灌注(OGD/R)诱导的PC12细胞活力的影响,活性氧(ROS)清除,抗炎,神经保护和抑制细胞凋亡。此外,氯化2,3,5-三苯基四唑染色,苏木精和伊红染色,尼氏染色,和免疫组织化学用于检测梗死面积,病理变化,大脑中动脉阻塞(MCAO)大鼠的Nissl小体和神经元蛋白表达。在细胞和动物实验中进行聚合酶链反应和蛋白质印迹以检测ER应激相关基因和蛋白质的表达水平。
结果:MD提取物在OGD/R模型下增强PC12细胞的活力,降低ROS和IL-6水平,MBP水平升高,抑制细胞凋亡。此外,MD改善了MCAO大鼠的梗死面积,增加了Nissl尸体的数量,和调节神经元蛋白水平,包括微管相关蛋白2(MAP-2),髓鞘碱性蛋白(MBP),胶质纤维酸性蛋白(GFAP),和神经丝200(NF200)。此外,MD可以调节氧化应激蛋白丙二醛(MDA)的表达水平,一氧化氮(NO),超氧化物歧化酶(SOD),和过氧化氢酶(CAT)。细胞和动物实验均表明,MD可以抑制ER应激相关蛋白(GRP78,ATF4,ATF6,CHOP)并减少细胞凋亡。
结论:本研究证实,MD提取物对CIR损伤的治疗机制是通过抑制氧化应激和ER应激途径,除了抑制细胞凋亡。
BACKGROUND: Melastoma dodecandrum Lour. (MD), a traditional Chinese medicine used by the She ethnic group, has been used to treat cerebral ischemia-reperfusion (CIR) injury due to its efficacy in promoting blood circulation and removing blood stasiss; however, the therapeutic effects and mechanisms of MD in treating CIR injury remain unclear.
OBJECTIVE: To investigate the protective effects of MD on CIR injury, in addition to its impact on oxidative stress, endoplasmic reticulum (ER) stress, and cell apoptosis.
METHODS: The research was conducted using both cell experiments and animal experiments. The CCK-8 method, immunofluorescence staining, and flow cytometry were used to analyze the effects of MD-containing serum on oxygen-glucose deprivation/reperfusion (OGD/R)-induced PC12 cell viability, reactive oxygen species (ROS) clearance, anti-inflammatory, neuroprotection and inhibition of apoptosis. Furthermore, 2,3,5-Triphenyl tetrazolium chloride staining, hematoxylin and eosin staining, Nissl staining, and immunohistochemistry were used to detect infarct size, pathological changes, Nissl corpuscula and neuronal protein expression in middle cerebral artery occlusion (MCAO) rats. Polymerase chain reaction and Western Blotting were conducted in cell and animal experiments to detect the expression levels of ER stress-related genes and proteins.
RESULTS: The MD extract enhanced the viability of PC12 cells under OGD/R modeling, reduced ROS and IL-6 levels, increased MBP levels, and inhibited cell apoptosis. Furthermore, MD improved the infarct area in MCAO rats, increased the number of Nissl bodies, and regulated neuronal protein levels including Microtubule-Associated Protein 2 (MAP-2), Myelin Basic Protein (MBP), Glial Fibrillary Acidic Protein (GFAP), and Neurofilament 200 (NF200). Additionally, MD could regulate the expression levels of oxidative stress proteins malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase (SOD), and catalase (CAT). Both cell and animal experiments demonstrated that MD could inhibit ER stress-related proteins (GRP78, ATF4, ATF6, CHOP) and reduce cell apoptosis.
CONCLUSIONS: This study confirmed that the therapeutic mechanism of the MD extract on CIR injury was via the inhibition of oxidative stress and the ER stress pathway, in addition to the inhibition of apoptosis.