School of Animal Engineering, Xuzhou Vocational College of Bioengineering, Xuzhou, China.
School of Traditional Chinese Pharmacy, China Pharmaceutical University, Jiangsu Taixing, Nanjing, China.
Indian J Pharmacol. 2020 Mar-Apr;52(2):108-116. doi: 10.4103/ijp.IJP_400_18. Epub 2020 Jun 3.
We investigate the protective effect of L. (CTL, also known as Honghua in China or Safflower) on cerebral ischemia-reperfusion and explored the possible mechanisms on regulating apoptosis and matrix metalloproteinases (MMPs). High-performance liquid chromatography method with diode array detection analysis was established to analyze the components of CTL. Middle cerebral artery occlusion rats model was established to evaluate Neurological Function Score and hematoxylin-eosin staining, as well as triphenyltetrazolium was used to examine the infarction area ratio. Transferase-mediated dUTP nick-end labeling was performed for the apoptosis. Apoptosis-related factors, including B-cell lymphoma-2 (Bcl-2), Bax and Caspase3, and MMPs-related MMP2, MMP9, tissue inhibitor of metalloproteinases 1 (TIMP1) in ischemic brain, were assayed by Western blot, reverse transcription polymerase chain reaction, and immunohistochemistry. The data showed that CTL (2, 4 g crude drug/kg/d) treatment could significantly reduce the ischemic damage in brain tissue and improve a significant neurological function score. In addition, CTL could also attenuate apoptosis degree of brain tissues and regulate Bcl-2, Bax, and Caspase 3 and also have a significant decrease on MMP-9 expression, followed by a significant increase of TIMP1 protein expression. These findings indicated that regulation of CTL on apoptosis and MMPs contributed to its protective effect on ischemia/reperfusion injury.
我们研究了 L.(CTL,在中国也称为红花或红花)对脑缺血再灌注的保护作用,并探讨了调节细胞凋亡和基质金属蛋白酶(MMPs)的可能机制。采用二极管阵列检测分析的高效液相色谱法分析 CTL 的成分。建立大脑中动脉闭塞大鼠模型,评估神经功能评分和苏木精-伊红染色,并使用三苯基四唑测定梗死面积比。采用转移酶介导的 dUTP 缺口末端标记法进行细胞凋亡检测。采用 Western blot、逆转录聚合酶链反应和免疫组织化学法检测缺血脑组织中与细胞凋亡相关的因子,包括 B 细胞淋巴瘤-2(Bcl-2)、Bax 和 Caspase3,以及与 MMPs 相关的 MMP2、MMP9 和金属蛋白酶组织抑制剂 1(TIMP1)。结果表明,CTL(2、4 g 生药/kg/d)治疗可显著减轻脑组织缺血损伤,显著提高神经功能评分。此外,CTL 还可减轻脑组织的凋亡程度,调节 Bcl-2、Bax 和 Caspase3,并显著降低 MMP-9 表达,同时 TIMP1 蛋白表达显著增加。这些发现表明,CTL 对细胞凋亡和 MMPs 的调节有助于其对缺血/再灌注损伤的保护作用。