Institute of Traditional Chinese Medicine Resources, College of Pharmaceutical Science, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 311400, PR China.
Institute of Traditional Chinese Medicine Resources, College of Pharmaceutical Science, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 311400, PR China.
J Ethnopharmacol. 2020 Dec 5;263:113168. doi: 10.1016/j.jep.2020.113168. Epub 2020 Jul 27.
In vitro cultured calculus bovis (ICCB), which is produced based on the formation mechanism of bovine gallstones, is used to replace the natural bezoar. It has been used in the clinic to treat brain diseases, including stroke, Alzheimer's disease and depression.
ICCB is used to treat encephalopathy in the clinic. We explored the effects of ICCB on cerebral ischaemia-reperfusion injury (CIRI) and the potential associated mechanisms.
Rats were subjected to middle cerebral artery occlusion for 90 min, followed by 24 h of reperfusion, after being given different concentrations of ICCB once a day for 3 days. Subsequently, the neurological scores, brain oedema and volume of cerebral infarction were measured, and the histopathological changes in the cortex neurons were observed by haematoxylin and eosin staining (H&E). Apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL). Ultrastructural changes in the mitochondria of the cortex were assessed by transmission electron microscopy (TEM). The apoptosis-related proteins Bax, Bcl-2, caspase-9, caspase-3, Mito-Cyt C and Cyto-Cyt C were detected by Western blotting.
Compared with those in the control group, the neurological scores, the volumes of cerebral infarction, and the brain water contents were significantly decreased in the ICCB groups at doses of 50 and 100 mg/kg. The ICCB treatment effectively decreased the neuronal apoptosis resulting from the CIRI-induced neuron injury. In addition, the histopathological damage and the mitochondria ultrastructure injury were partially improved in the CIRI rats after ICCB treatment. Western blotting analysis indicated that ICCB significantly decreased the expression of Bax, caspase-9, caspase-3 and Cyto-Cyt C protein levels while increasing the expression of Bcl-2 and Mito-Cyt C protein levels.
The ICCB protected against CIRI by suppressing the mitochondria-mediated apoptotic signalling pathway.
基于牛胆结石形成机制生产的体外培养牛黄（ICCB）已被用于替代天然牛黄，用于治疗中风、阿尔茨海默病和抑郁症等脑部疾病。
ICCB 用于临床治疗脑病。我们探讨了 ICCB 对脑缺血再灌注损伤（CIRI）的影响及其潜在的相关机制。
大鼠大脑中动脉闭塞 90 分钟后，再灌注 24 小时，然后每天给予不同浓度的 ICCB 一次，连续 3 天。随后，测量神经功能评分、脑水肿和脑梗死体积，通过苏木精和伊红染色（H&E）观察皮质神经元的组织病理学变化。通过末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记（TUNEL）测定细胞凋亡。通过透射电子显微镜（TEM）评估皮质线粒体的超微结构变化。通过 Western blot 检测凋亡相关蛋白 Bax、Bcl-2、caspase-9、caspase-3、Mito-Cyt C 和 Cyto-Cyt C。
与对照组相比，50 和 100mg/kg ICCB 组的神经功能评分、脑梗死体积和脑水含量均显著降低。ICCB 治疗有效减少了 CIRI 诱导神经元损伤引起的神经元凋亡。此外，ICCB 治疗部分改善了 CIRI 大鼠的组织病理学损伤和线粒体超微结构损伤。Western blot 分析表明，ICCB 显著降低了 Bax、caspase-9、caspase-3 和 Cyto-Cyt C 蛋白水平的表达，同时增加了 Bcl-2 和 Mito-Cyt C 蛋白水平的表达。
ICCB 通过抑制线粒体介导的凋亡信号通路来保护 CIRI。
