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天麻芎苓止眩片对血管内皮细胞自噬的影响及机制
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| 篇名： | 天麻芎苓止眩片对血管内皮细胞自噬的影响及机制 |
| TITLE： | Effects and mechanism of Tianma xiongling zhixuan tablet on autophagy of vascular endothelial cells |
| 摘要： | 目的 探究天麻芎苓止眩片对大鼠血管内皮细胞自噬的影响及潜在机制。方法将大鼠主动脉内皮细胞（RAECs）分为正常组、模型组、空白血清组、中药含药血清组、自噬阻断剂组、自噬激动剂组、中药联合自噬激动剂组。除正常组外，其余各组分别加入10μg/mL脂多糖干预24ｈ建立RAECs炎症损伤模型。空白血清组加入10%空白血清，中药含药血清组加入10%天麻芎苓止眩片含药血清，自噬阻断剂组加入20μmol/LPD98059，自噬激动剂组加入50μmol/LHonokiol，中药联合自噬激动剂组加入10%天麻芎苓止眩片含药血清和50μmol/LHonokiol。观察各组RAECs形态学特征，检测各组细胞活力、上清液中内皮素1（ET-1）和一氧化氮（NO）水平、线粒体活性氧水平、线粒体膜电位，以及PTEN诱导激酶1（PINK1）、E3泛素连接酶Parkin、选择性自噬接头蛋白62（p62）、微管相关蛋白1轻链3（LC3）的蛋白表达。结果与模型组比较，自噬阻断剂组和中药含药血清组的ET-1水平、线粒体活性氧水平及PINK1、Parkin、LC3蛋白的相对表达量均显著降低或下调（P＜0.05或P＜0.01），细胞存活率（仅自噬阻断剂组）、NO水平、线粒体膜电位、p62蛋白的相对表达量均显著升高或上调（P＜0.05或P＜0.01），RAECs病理损伤改善明显，细胞数量显著增加，恢复典型的铺路石样特点；自噬激动剂组的ET-1水平、线粒体活性氧水平及Parkin和LC3蛋白的相对表达量均显著升高或上调（P＜0.05或P＜0.01），细胞存活率显著降低（P＜0.05），病理损伤加重。与自噬激动剂组比较，中药联合自噬激动剂组细胞存活率、p62蛋白相对表达量均显著升高或上调（P＜0.05或P＜0.01），ET-1水平及PINK1、Parkin、LC3蛋白的相对表量均显著下调（P＜0.01），RAECs形态结构受损被一定程度逆转。结论天麻芎苓止眩片可通过调控线粒体自噬来保护血管内皮功能，其作用机制可能与调控PINK1/Parkin信号通路、抑制线粒体自噬有关。 |
| ABSTRACT： | OBJECTIVE To explore the effects of Tianma xiongling zhixuan tablet on autophagy in vascular endothelial cells of rats and its potential mechanism. METHODS The rat aortic endothelial cells （RAECs） were divided into normal group， model group， blank serum group， traditional Chinese medicine （TCM） medicated serum group， autophagy blocker group， autophagy agonist group， and TCM combined with autophagy agonist group. Except for normal group， other groups were given 10 μg/mL lipopolysaccharide for 24 hours to induce RAECs inflammation injury model. Blank serum group was treated with 10% blank serum； TCM medicated serum group received 10% medicated serum derived from Tianma xiongling zhixuan tablet； autophagy blocker group was treated with 20 μmol/L of PD98059； autophagy agonist group was administered 50 μmol/L Honokiol. Lastly， the TCM combined with autophagy agonist group was given both 10% medicated serum derived from Tianma xiongling zhixuan tablet and 50 μmol/L Honokiol. The morphological characteristics of RAECs in each group were observed. The cell viability of each group， the contents of endothelin-1 （ET-1） and nitric oxide （NO）， mitochondrial reactive oxygen species， mitochondrial membrane potential， and the expression levels of PTEN-induced kinase 1 （PINK1）， Parkin， ubiquitin-binding protein （p62）， and microtubule-associated protein 1 light chain 3 （LC3） were detected. RESULTS Compared with model group， the levels of ET-1， mitochondrial reactive oxygen species， and the relative expressions of PINK1， Parkin， and LC3 proteins in the autophagy blocker group and TCM medicated serum group were decreased or down-regulated significantly （P＜0.05 or P＜0.01）； the cell viability rate （only autophagy blocker group）， NO level， mitochondrial membrane potential， and the E-mail：46164660@qq.com relative expression level of p62 protein were increased or up-regulated significantly （P＜0.05 or P＜0.01）； the pathological damage of RAECs was significantly improved， the number of cells increased significantly， and the typical paving stone-like characteristics were restored. The levels of ET-1， mitochondrial reactive oxygen species， and the relative expression levels of Parkin and LC3 proteins in the autophagy agonist group were increased or up-regulated significantly （P＜0.05 or P＜0.01）， while cell viability rate was decreased significantly （P＜0.05）， the damage of RAECs was aggravated. Compared with the autophagy agonist group， the cell viability rate and the relative expression level of p62 protein in TCM combined autophagy agonist group were increased or up-regulated significantly （P＜0.05 or P＜0.01）， while the levels of ET-1， the relative expression levels of PINK1， Parkin， and LC3 proteins were down-regulated significantly （P＜ 0.01）， the damage of RAECs was reversed to a certain extent. CONCLUSIONS Tianma xiongling zhixuan tablet protects vascular endothelial function by regulating mitochondrial autophagy， the mechanism of which may be associated with the regulation of PINK1/Parkin signaling pathway and the inhibition of mitochondrial autophagy. |
| 期刊： | 2025年第36卷第14期 |
| 作者： | 雍苏南；房赤；龙远雄；李萍；谢小兵 |
| AUTHORS： | YONG Sunan，FANG Chi，LONG Yuanxiong，LI Ping，XIE Xiaobing |
| 关键字： | 天麻芎苓止眩片；血管内皮细胞损伤；线粒体自噬；PINK1/Parkin信号通路 |
| KEYWORDS： | Tianma xiongling zhixuan tablet； vascular endothelial cell injury； mitochondrial autophagy； PINK1/Parkin |
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