背景:补阳还五汤在中国临床上广泛用于缺血性脑血管病的治疗和预防。本研究旨在探讨BHD对缺血性卒中(IS)的影响及其潜在机制。
方法:建立大脑中动脉闭塞(MCAO)大鼠模型和氧糖剥夺复氧(OGD/R)大鼠脑微血管内皮细胞(RBMVEC)模型。MCAO手术后计算脑梗死大小和神经评分。伊文思蓝用于测量血脑屏障(BBB)通透性。进行细胞计数试剂盒-8(CCK-8)和TUNEL测定以评估RBMVEC的细胞活力和凋亡。采用双荧光素酶报告基因分析凋亡相关基因的转录活性。
结果:结果显示较高的梗死体积,神经学评分,BHD治疗后MCAO组大鼠BBB通透性降低。药物血清(DS)处理对正常的RBMVECs细胞活力和细胞凋亡没有影响。此外,DS处理降低了乳酸的产生,葡萄糖摄取,正常和OGD/R诱导的RBMVECs的细胞外酸化率。DS处理下调了泛赖氨酸乳酸化(kla)的蛋白质水平,组蛋白H3赖氨酸18(H3K18la),以及OGD/R处理的RBMVECs中凋亡蛋白酶活化因子-1(Apaf-1)的转录。此外,Apaf-1过表达降低了OGD/R处理的RBMVECs的细胞活力并增加了细胞凋亡和糖酵解活性。
结论:总之,BHD通过抑制pan-kla和H3K18la蛋白水平和Apaf-1转录活性抑制糖酵解和细胞凋亡,从而限制了IS的发展。
BACKGROUND: Buyang Huanwu Decoction (BHD) is widely used in Chinese clinical practice for the treatment and prevention of ischemic cerebral vascular diseases. This study was designed to investigate the effects of BHD on ischemic stroke (IS) and its underlying mechanism.
METHODS: The middle cerebral artery occlusion (
MCAO) rat model and oxygen-glucose deprivation and reoxygenation (OGD/R) rat brain microvascular endothelial cell (RBMVEC) models were established. Brain infarction size and neurological score were calculated following
MCAO surgery. Evans blue was used to measure blood brain barrier (BBB) permeability. Cell counting kit-8 (CCK-8) and TUNEL assays were performed to evaluate the cell viability and apoptosis of RBMVECs. Dual-luciferase reporter assay was used to analyze the transcriptional activities of apoptosis-related genes.
RESULTS: Results showed that higher infarction volume, neurological scores, and BBB permeability in the
MCAO group rats were reduced after BHD treatment. Drug serum (DS) treatment had no impact on the normal RBMVECs\' cell viability and cell apoptosis. Besides, DS treatment decreased the lactate production, glucose uptake, and extracellular acidification rate in normal and OGD/R-induced RBMVECs. DS treatment downregulated the protein levels of pan-lysine lactylation (kla), histone H3 lysine 18 lactylation (H3K18la), and the transcriptional of apoptotic protease activating factor-1 (Apaf-1) in OGD/R-treated RBMVECs. In addition, Apaf-1 overexpression decreased cell viability and increased apoptosis and glycolysis activity of OGD/R-treated RBMVECs.
CONCLUSIONS: In summary, BHD inhibited glycolysis and apoptosis via suppressing the pan-kla and H3K18la protein levels and the Apaf-1 transcriptional activity, thus restraining the progression of IS.