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龚翠兰,马强,杨仁义,王智槟,周德生.基于JAK2/STAT3通路探讨活血荣络方对OGD/R后BMEC的干预作用及机制[J].湖南中医药大学学报,2024,44(3):350-356[点击复制] |
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| 基于JAK2/STAT3通路探讨活血荣络方对OGD/R后BMEC的干预作用及机制 |
| 龚翠兰,马强,杨仁义,王智槟,周德生 |
| (湖南中医药大学附属常德医院, 湖南 常德 415000;湖南中医药大学第一附属医院, 湖南 长沙 410007;湖南省中西医结合医院, 湖南 长沙 410006) |
| 摘要: |
| 目的 探讨活血荣络方激活Janus酪氨酸激酶2/信号转导和转录激活剂3（Janus kinase 2/signal transducer and activator of transcription 3， JAK2/STAT3）通路对氧糖剥夺/复氧（oxygen-glucose deprivation/reperfusion， OGD/R）后脑微血管内皮细胞（brain microvascular endothelial cell， BMEC）的干预作用和机制。方法 培养大鼠BMEC（bEnd.3细胞），建立OGD/R模型；将细胞随机分为正常组（10%空白血清）、模型组（10%空白血清）、活血荣络方组（10%活血荣络方含药血清）、丁苯酞组（10%丁苯酞含药血清）、Stattic组（10%空白血清+10 μmol/mL Stattic）、联用组（10%活血荣络方含药血清+10 μmol/mL Stattic），并分别给予相应的药物干预24 h。采用CCK-8法检测活血荣络方对OGD/R后bEnd.3细胞活性的影响；采用划痕实验和Transwell迁移实验分别检测活血荣络方对OGD/R后bEnd.3细胞划痕愈合和细胞迁移的影响；采用Western blot法检测bEnd.3细胞中JAK2、p-JAK2、STAT3、p-STAT3、血管内皮细胞生长因子A（vascular endothelial growth factor A，VEGFA）蛋白表达水平。结果 与正常组比较，模型组bEnd.3细胞存活率、细胞划痕愈合率、细胞迁移数显著降低（P<0.01），JAK2、p-JAK2、STAT3、p-STAT3、VEGFA的表达量升高（P<0.05或P<0.01）；与模型组比较，活血荣络方组bEnd.3细胞的存活率、细胞划痕愈合率、细胞迁移数显著升高（P<0.01），JAK2、p-JAK2、p-STAT3、STAT3、VEGFA的表达量升高（P<0.05或P<0.01）；与Stattic组比较，联用组bEnd.3细胞的存活率、细胞划痕愈合率、细胞迁移数显著升高（P<0.05或P<0.01），STAT3、VEGFA的表达量升高（P<0.05）。结论 活血荣络方可能激活JAK2/STAT3信号通路并诱导内皮细胞的增殖、迁移和存活，激活VEGFA表达并减轻脑缺血再灌注损伤，具有神经保护作用。 |
| 关键词: 缺血性脑卒中 活血荣络方 JAK2/STAT3通路 氧糖剥夺/复氧 脑微血管内皮细胞 |
| DOI：10.3969/j.issn.1674-070X.2024.03.002 |
| 投稿时间：2023-08-19 |
| 基金项目:国家自然科学基金项目（8210151338）；湖南省自然科学基金项目（2022JJ30088，2023JJ40512，2021JJ30521，2021JJ40424）；湖南省卫生健康委员会基金项目（202203074864）。 |
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| Intervention and mechanism of Huoxue Rongluo Formula on BMEC after OGD/R based on JAK2/STAT3 pathway |
| GONG Cuilan,MA Qiang,YANG Renyi,WANG Zhibing,ZHOU Desheng |
| (Changde Hospital of Hunan University of Chinese Medicine, Changde, Hunan 415000, China;The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Hunan Provincial Hospital of Integrated Chinese and Western Medicine, Changsha, Hunan 410006, China) |
| Abstract: |
| Objective To investigate the intervention and mechanism of Huoxue Rongluo Formula (HXRLF) on brain microvascular endothelial cell (BMEC) after oxygen-glucose deprivation/reoxygenation (OGD/R) by activating Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) pathway. Methods Rat BMEC (bEnd.3 cell) was cultivated and an OGD/R model was established. The cells were randomized into normal (10% blank serum), model (10% blank serum), HXRLF (10% HXRLF containing serum), butylph-thalide (10% butylphthalide containing serum), Stattic (10% blank serum+10 μmol/mL Stattic), and combination (10% HXRLF serum+10% μmol/mL Stattic) groups, and were given corresponding drug interventions for 24 h, respectively. CCK-8 assay was used to determine the effects of HXRLF on the activity of bEnd.3 cells after OGD/R. Wound healing assay and Transwell migration assay were used to test the effects of HXRLF on scratch healing and cell migration ability of bEnd.3 cells after OGD/R; the protein expression levels of JAK2, p-JAK2, STAT3, p-STAT3, and VEGFA in bEnd.3 cells were determined by Western-blot. Results Compared with the normal group, the survival rate, scratch healing rate, and cell migration number of bEnd.3 cells in the model group were significantly lower (P<0.01), while the expression levels of JAK2, p-JAK2, STAT3, p-STAT3, and VEGFA were higher (P<0.05 or P<0.01). Compared with the model group, the survival rate, scratch healing rate, and migration number of bEnd.3 cells in the HXRLF group were significantly higher (P<0.01), and the expression levels of JAK2, p-JAK2, p-STAT3, STAT3, and VEGFA were significantly higher (P<0.05 or P<0.01). Compared with the Stattic group, the survival rate, scratch healing rate, and cell migration number of bEnd.3 cells in the combination group were significantly higher (P<0.05 or P<0.01), and the expression levels of STAT3 and VEGFA were higher (P<0.05). Conclusion HXRLF may activate the JAK2/STAT3 signaling pathway, induce endothelial cell proliferation, migration, and survival, activate VEGFA expression, and alleviate cerebral ischemia-reperfusion injury, thus exerting neuroprotective effects. |
| Key words: ischemic stroke Huoxue Rongluo Formula Janus kinase 2/signal transducer and activator of transcription 3 pathway oxygen-glucose deprivation/reoxygenation brain microvascular endothelial cell |
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