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目的:研究太子参对去甲肾上腺素(NE)诱发原代培养心肌细胞损伤保护作用的活性部位。方法:采用系统溶剂分离制备提取太子参石油醚,乙酸乙酯,正丁醇及水层部位。取1～3 d的SD大鼠乳鼠制备原代培养心肌细胞,以1×105/mL的细胞密度接种于96孔培养板,以相当于生药0.1～0.25 g.mL-1的太子参不同提取部位作用于原代培养心肌细胞30 min后,以1μmol.L-1NE作用于心肌细胞24 h复制细胞损伤模型,MTT法分析不同提取部位太子参的保护作用。结果:正丁醇和水层提取部位对NE诱导原代培养心肌细胞损伤吸光度(A570)降低具有显著的提高作用;进一步分析提示0.1 g.mL-1和0.25 g.mL-1太子参25%乙醇洗脱正丁醇部位和多糖物质群对NE诱导心肌细胞A570的降低保护作用显著(与模型组比较,差异显著,P<0.01,P<0.05)。结论:初步的活性筛选发现太子参的正丁醇部位和水层部位对NE诱导的心肌细胞损伤具有保护作用,进一步的活性追踪确定正丁醇部位经25%乙醇洗脱物质群及水层中的粗多糖为太子参防治NE诱导的心肌细胞损伤的主要活性物质群。
Objective: To study the protective effect of Taizishen on the injury of cardiomyocytes induced by norepinephrine (NE). Methods: The petroleum ether, ethyl acetate, n-butanol and water layer of Pseudostellaria heterophylla were prepared by systematic solvent separation. Primary rat cardiomyocytes were prepared from 1 to 3 days old SD rat pups and inoculated into 96-well plates at a cell density of 1 × 10 5 / mL to give different doses of Radix Pseudostellariae Radix which is equivalent to 0.1 to 0.25 g.mL-1 of crude drug After 30 min of primary cultured cardiomyocytes, the cells were treated with 1 μmol·L-1 NE for 24 h. The MTT assay was used to analyze the protective effect of Radix Pseudostellariae heterophyllae on different extracts. Results: The extracts of n-butanol and aqueous extracts had a significant effect on the decrease of absorbency (A570) induced by NE-induced primary cultured cardiomyocytes. Further analysis suggested that 0.1 g.mL-1 and 0.25 g.mL-1 Pseudostellaria heterophylla 25% ethanol Elution of n-butanol and polysaccharide groups had a significant protective effect on the NE-induced A570 myocardial cell (compared with the model group, the difference was significant, P <0.01, P <0.05). CONCLUSION: The results of preliminary bioassay showed that n-butanol fraction and water fraction of Pseudostellaria heterophylla had a protective effect on NE-induced injury of cardiomyocytes. Further activity was followed up to determine the n-butanol fraction eluted by 25% Of crude polysaccharide is the main active substance group that Pseudostellaria ginseng can prevent NE-induced cardiomyocyte injury.