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通络救脑注射液通过抑制 MIP-1β 表达对脑微血管内皮细胞缺氧缺糖损伤的保护作用

Tong Luo Jiu Nao injection, a traditional Chinese medicinal preparation, inhibits MIP-1β expression in brain microvascular endothelial cells injured by oxygen-glucose deprivation.

机构信息

Institute of Basic Theory, China Academy of Chinese Medical Sciences, 16 Dong Zhi Men Nei Nan Xiao Jie, Dong Cheng District, Beijing 100700, China.

出版信息

J Ethnopharmacol. 2012 May 7;141(1):151-7. doi: 10.1016/j.jep.2012.02.013. Epub 2012 Feb 17.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Tong Luo Jiu Nao injection (TLJN), a modern Chinese formula based on Traditional Chinese Medicine theory, has been used to treat ischemic stroke and vascular dementia. TLJN belongs to the ethnopharmacological family of medicines.

AIM OF THE STUDY

To investigate the protective effect of TLJN on oxygen-glucose deprivation (OGD) induced-injury of brain microvascular endothelial cells (BMECs).

MATERIALS AND METHODS

The model of OGD was established in the primarily cultured BMECs. TLJN was added to the OGD-injured BMECs for 6h. A series of assays were used to detect the effects of TLJN on: (i) MIP-1β content in BMECs conditioned media (CM) by ELISA; (ii) MIP-1β protein expression in BMECs by western blotting and immunocytochemistry; (iii) the expression of CCR5, receptor of MIP-1β, in BMECs by western blotting; (iv) the proliferative activity of microglial cells via the Cell Counting Kit-8 (CCK-8).

RESULTS

Our results showed that the OGD-injured BMECs presented with large amounts of secretion and expression of MIP-1β and up-regulation of CCR5. Also, the CM of OGD-injured BMECs remarkably increased the proliferative activity of microglial cells. The TLJN-treated BMECs exhibited a reduction of MIP-1β content in BMECs-CM and a down-regulation of MIP-1β and CCR5 expression. In addition, an inhibitory effect of CM of OGD-injured plus TLJN injection-treated BMECs on microglial proliferation was also found.

CONCLUSION

TLJN reduced the expression of MIP-1β and CCR5 in OGD-injured BMECs, and the CM of OGD-injured plus TLJN injection-treated BMECs inhibited the proliferative activity of microglial cells, suggesting the therapeutic potential of TLJN on ischemic cerebral vascular disease.

摘要

民族药理学相关性

通络救脑注射液(TLJN)是一种基于中医理论的现代中药配方,用于治疗缺血性中风和血管性痴呆。TLJN 属于民族药理学药物家族。

目的

研究 TLJN 对氧葡萄糖剥夺(OGD)诱导的脑微血管内皮细胞(BMEC)损伤的保护作用。

材料和方法

在原代培养的 BMECs 中建立 OGD 模型。TLJN 加入 OGD 损伤的 BMECs 中 6 小时。一系列实验用于检测 TLJN 对以下方面的影响:(i)通过 ELISA 检测 BMECs 条件培养基(CM)中 MIP-1β 的含量;(ii)通过 Western blot 和免疫细胞化学检测 BMECs 中 MIP-1β 的蛋白表达;(iii)通过 Western blot 检测 BMECs 中 MIP-1β 受体 CCR5 的表达;(iv)通过细胞计数试剂盒-8(CCK-8)检测小胶质细胞的增殖活性。

结果

我们的结果表明,OGD 损伤的 BMECs 大量分泌和表达 MIP-1β,并上调 CCR5。此外,OGD 损伤的 BMECs 的 CM 显著增加了小胶质细胞的增殖活性。TLJN 处理的 BMECs 表现出 BMECs-CM 中 MIP-1β 含量减少,MIP-1β 和 CCR5 表达下调。此外,还发现 OGD 损伤加 TLJN 注射处理的 BMECs 的 CM 对小胶质细胞增殖具有抑制作用。

结论

TLJN 降低了 OGD 损伤的 BMECs 中 MIP-1β 和 CCR5 的表达,OGD 损伤加 TLJN 注射处理的 BMECs 的 CM 抑制了小胶质细胞的增殖活性,提示 TLJN 对缺血性脑血管病具有治疗潜力。

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