Electroacupuncture Serum Protects Blood-brain Barrier Damage after Ischemic Stroke BY Regulating the Pericytes in vitro.

Hanrui Zhang, Hequn Lyv, Yaoting Feng, Yongjun Peng
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Abstract

Background: Electroacupuncture (EA) exerts a protective role in Blood-Brain Barrier (BBB) damage after ischemic stroke, but whether this effect involves the regulation of the pericytes in vitro is unclear.

Methods: The in vitro BBB models were established with brain microvascular endothelial cells (BMECs) and pericytes, and the co-cultured cells were randomly divided into three groups: the control group, oxygen-glucose deprivation/reoxygenation (OGD/R) group and EA group. OGD/R was performed to simulate cerebral ischemia-reperfusion in vitro. EA serum was prepared by EA treatment at the "Renzhong" (GV26) and "Baihui" (GV20) acupoints in middle cerebral artery occlusion/ reperfusion rats. Furthermore, the characteristics of BMECs and pericytes were identified with immunohistochemistry staining. The cell morphology of the BBB model was observed using an inverted microscope. The function of BBB was measured with transendothelial electrical resistance (TEER) and sodium fluorescein, and the viability, apoptosis, and migration of pericytes were detected by cell counting kit-8, flow cytometry, and Transwell migration assay.

Results: BMECs were positive staining for Factor-VIII, and pericytes were positive staining for the α-SMA and NG2. EA serum improved cell morphology of the BBB model increased TEER, and decreased sodium fluorescein in OGD/R condition. Besides, EA serum alleviated pericytes" apoptosis rate and migration number, and enhanced pericytes' viability rate in OGD/R condition.

Conclusion: EA serum protects against BBB damage induced by OGD/R in vitro, and this protection might be achieved by attenuating pericytes apoptosis and migration, as well as enhancing pericytes viability. The findings provided new evidence for EA as a medical therapy for ischemic stroke.

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电针血清通过体外调节周细胞保护缺血性脑卒中后的血脑屏障损伤
背景:电针(EA)对缺血性脑卒中后血脑屏障(BBB)损伤有保护作用,但这种作用是否涉及体外周细胞的调控尚不清楚:方法:利用脑微血管内皮细胞(BMECs)和周细胞建立体外 BBB 模型,并将共培养的细胞随机分为三组:对照组、氧-葡萄糖剥夺/复氧(OGD/R)组和 EA 组。OGD/R 是在体外模拟脑缺血再灌注。EA血清是通过EA治疗大脑中动脉闭塞/再灌注大鼠的 "人中"(GV26)和 "百会"(GV20)穴位制备的。此外,还通过免疫组化染色鉴定了BMECs和周细胞的特征。使用倒置显微镜观察 BBB 模型的细胞形态。用跨内皮电阻(TEER)和荧光素钠测量 BBB 的功能,用细胞计数试剂盒-8、流式细胞术和 Transwell 迁移试验检测周细胞的活力、凋亡和迁移:结果:BMECs的因子-VIII呈阳性染色,周细胞的α-SMA和NG2呈阳性染色。EA 血清改善了 BBB 模型的细胞形态,增加了 TEER,并降低了 OGD/R 条件下的荧光素钠。此外,EA血清还降低了OGD/R条件下周细胞的凋亡率和迁移数量,并提高了周细胞的存活率:结论:EA血清对体外OGD/R诱导的BBB损伤有保护作用,这种保护作用可能是通过减少周细胞凋亡和迁移以及提高周细胞存活率实现的。这些研究结果为EA作为缺血性中风的一种医学疗法提供了新的证据。
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