Zhuge Lujie, Fang Yan, Jin Huaqian, Li Lin, Yang Yan, Hu Xiaowei, Chu Lisheng
College of Pharmaceutical Science, Zhejiang Chinese Medical University, Hangzhou 310053, China.
College of Basic Medical Sciences, Zhejiang Chinese Medical University, Hangzhou 310053, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2020 Dec 25;49(6):687-696. doi: 10.3785/j.issn.1008-9292.2020.12.03.
To investigate the mechanism of Chinese medicine Buyang Huanwu decoction (BYHWD) promoting neurogenesis and angiogenesis in ischemic stroke rats.
Male SD rats were randomly divided into sham operation group, model group, BYHWD group, antagonist group and antagonist control group with 14 rats in each. Focal cerebral ischemia was induced by occlusion of the right middle cerebral artery for 90 min with intraluminal filament and reperfusion for 14 d in all groups except sham operation group. BYHWD (13 g/kg) was administrated by gastrogavage in BYHWD group, antagonist group and antagonist control group at 24 h after modeling respectively, and BrdU (50 mg/kg) was injected intraperitoneally in all groups once a day for 14 consecutive days. miR-199a-5p antagomir or NC (10 nmol) was injected into the lateral ventricle at d5 after ischemia in antagonist and antagonist control groups, respectively. The neurological deficits were evaluated by the modified neurological severity score (mNSS) and the corner test, and the infract volume was measured by toluidine blue staining. Neurogenesis and angiogenesis were detected by immunofluorescence double labeling method. The expression level of miR-199a-5p was tested by real-time RT-PCR, and the protein expressions of vascular endothelial growth factor (VEGF) and brain-derived neurotrophic factor (BDNF) were determined by Western blotting.
BYHWD treatment significantly promoted the recovery of neurological function (<0.05 or <0.01), reduced the infarct volume (<0.05), increased the number of BrdU/DCX, BrdU/NeuN and BrdU/vWF cells (all <0.01), upregulated the expression of miR-199a-5p (<0.01), and increased the protein expression of VEGF and BDNF at d14 after cerebral ischemia (all <0.05). The above effects were reversed by intracerebroventricular injection of miR-199a-5p antagomir.
Buyang Huanwu decoction promotes neurogenesis and angiogenesis in rats with cerebral ischemia, which may be related to increased protein expression of VEGF and BDNF through upregulating miR-199a-5p.
探讨中药补阳还五汤（BYHWD）促进缺血性脑卒中大鼠神经发生和血管生成的机制。
雄性SD大鼠随机分为假手术组、模型组、补阳还五汤组、拮抗剂组和拮抗剂对照组，每组14只。除假手术组外，其余各组均采用线栓法阻断右侧大脑中动脉90分钟，再灌注14天制备局灶性脑缺血模型。补阳还五汤组、拮抗剂组和拮抗剂对照组在造模后24小时分别给予补阳还五汤（13 g/kg）灌胃，所有组连续14天每天腹腔注射BrdU（50 mg/kg）。拮抗剂组和拮抗剂对照组在缺血后第5天分别向侧脑室注射miR-199a-5p拮抗剂或NC（10 nmol）。采用改良神经功能缺损评分（mNSS）和转角试验评估神经功能缺损，通过甲苯胺蓝染色测量梗死体积。采用免疫荧光双标法检测神经发生和血管生成。通过实时RT-PCR检测miR-199a-5p的表达水平，采用Western blotting法检测血管内皮生长因子（VEGF）和脑源性神经营养因子（BDNF）的蛋白表达。
补阳还五汤治疗显著促进神经功能恢复（<0.05或<0.01），减小梗死体积（<0.05），增加BrdU/DCX、BrdU/NeuN和BrdU/vWF细胞数量（均<0.01），上调miR-199a-5p表达（<0.01），并增加脑缺血后第14天VEGF和BDNF的蛋白表达（均<0.05）。脑室内注射miR-199a-5p拮抗剂可逆转上述作用。
补阳还五汤可促进脑缺血大鼠的神经发生和血管生成，其机制可能与上调miR-199a-5p增加VEGF和BDNF蛋白表达有关。
