Beijing University of Chinese Medicine, No 11, Bei San Huan Dong Lu, Chao Yang District, Beijing, 100029, China.
China Academy of Chinese Medical Sciences Xiyuan Hospital, No1, Xiyuan Playground, Hai Dian District, Beijing, 100091, China.
J Pharmacol Sci. 2019 Mar;139(3):143-150. doi: 10.1016/j.jphs.2018.12.009. Epub 2019 Jan 22.
The aim of the present study was to examine the neuroprotective effect of Qingnao dripping pills (QNDP), especially the infiltration of neutrophils and macrophages, as well as the mitogen-activated protein kinase (MAPK) signal pathway. Adult male Sprague-Dawley rats were randomized to three groups: sham, MCAO, and QNDP. After 24 h of ischemia and reperfusion, neurological deficit scores and infarct volume were measured. Macrophages and neutrophil infiltration in the ischemic brain were respectively determined with CD68 and MPO immunofluorescence and western blot. The proteins involved in MAPK signaling (SAPK/JNK, P-SAPK/JNK, p38, P-p38, ERK1/2, and P-ERK1/2) were measured by western blotting. In vitro ischemic paradigm (oxygen-glucose deprivation) was performed in SH-SY5Y cells to evaluate the effects of QNDP. The viability and death ration of cells induced OGD/R was measured by MTT and LDH assay. The proteins involved in MAPK signaling were measured by western blotting. The results showed that QNDP treatment significantly improved the neurological deficit scores and reduced infarct size. In addition, QNDP treatment inhibited the number of CD68- and MPO-positive cells in the ischemic brain. It inhibited the MAPK signaling pathway in the ischemic brain and SH- SY5Y cells induced OGD/R.
本研究旨在探讨清脑滴丸（QNDP）的神经保护作用，特别是其对中性粒细胞和巨噬细胞浸润以及丝裂原活化蛋白激酶（MAPK）信号通路的影响。成年雄性 Sprague-Dawley 大鼠随机分为三组：假手术组、MCAO 组和 QNDP 组。缺血再灌注 24 h 后，测定神经功能缺损评分和梗死体积。用 CD68 和 MPO 免疫荧光和 Western blot 分别测定缺血脑组织中巨噬细胞和中性粒细胞的浸润。通过 Western blot 测定 MAPK 信号通路（SAPK/JNK、P-SAPK/JNK、p38、P-p38、ERK1/2 和 P-ERK1/2）中的蛋白。在 SH-SY5Y 细胞中进行体外缺血模型（氧葡萄糖剥夺），以评估 QNDP 的作用。通过 MTT 和 LDH 测定法测定 OGD/R 诱导的细胞活力和死亡率。通过 Western blot 测定 MAPK 信号通路中的蛋白。结果表明，QNDP 治疗可显著改善神经功能缺损评分，减少梗死体积。此外，QNDP 治疗可抑制缺血脑组织中 CD68 和 MPO 阳性细胞的数量。它抑制了缺血脑组织和 SH-SY5Y 细胞诱导的 OGD/R 中的 MAPK 信号通路。
