川芎嗪-原儿茶酸络合物的合成和其抗缺血性脑损伤作用初步研究

Synthesis and structure identification of Tetramethylpyrazine-Protocatechuic acid and effects on hypoxic-ischemic brain damage

目的:以川芎嗪、原儿茶酸为前体原料,运用药物拼合理论,合成川芎嗪-原儿茶酸络合物,并对其进行初步体内外缺血性脑损伤活性研究。方法:以川芎嗪(Tetramethylpyrazine,TMP)和原儿茶酸(Protocatechuic acid,PCA)为原料合成川芎嗪-原儿茶酸络合物(Tetramethylpyrazine-Protocatechuic acid,TP),运用UV、1H-NMR、13C-NMR、MS、X-Ray确定该化合物的结构。通过①鸡胚绒毛尿囊膜血管生成(Chicken chorioallantoic membrane,CAM)模型观察TP对新生血管生长的影响;②MTT法测定TP对原代培养的正常胎鼠皮层神经细胞、谷氨酸(Glutamate,Glu)和NO损伤神经细胞模型的影响;③TP对小鼠断头张口呼吸时间的影响等实验初步观察其对缺血性脑损伤的拮抗作用。结果:确定TMP和PCA两个分子片断之间借助一个O-H…O氢键和一个O-H…N氢键连接;TP对新生血管的生长具有显著的促进作用,改善Glu、NO对神经细胞的损伤,并能显著延长小鼠断头后张口呼吸时间。结论:由TMP和PCA拼合而成的TP对缺血性脑损伤具有明显改善作用,显示了其良好的新药开发研究前景。

TP is obtained on the basis of combination theory with（Tetramethylpyrazine,TMP）and（Protocatechuic acid,PCA）as the starting compounds which are 2 of biologically active ingredients from Ligusticum wallichii Franch and Radix Salviae Miltiorrhizae.To get exact structure of TP,we used by UV、1H-NMR、13C-NMR、MS and X-Ray.Previous studies have shown that NO and Glutamate（Glu） possess the ability to induce a series of raction after hypoxic-ischemic brain damage,and our results proved that TP significantly prevented the NO and Glu-induced reduction in cell survival via 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assay.In addition,TP significantly increase the proliferation of microvessels on experiment of CAM and also could dramatically prolong the gasping duration after decapitation in mice.Taken together,these results suggest that TP may be a potential ingredient for the hypoxic-ischemic brain damage and show a great prospects for new drug development.