侧柏炭不同溶剂提取物对LPS诱导的人脐静脉内皮细胞损伤的保护作用

Protective effect of different solvent extracts from Platycladi Cacumen Carbonisatum on LPS-induced human umbilical vein endothelial cells damage

目的:研究侧柏炭各溶剂提取物对脂多糖(LPS)诱导的人脐静脉内皮细胞(HUVECs)损伤的保护作用,探讨侧柏炭保护血管内皮细胞的有效提取物及其可能作用物质。方法:体外培养HUVECs,采用LPS诱导制备人脐静脉内皮细胞损伤模型。采用MTT比色法测定细胞活力、黄嘌呤氧化酶法测定细胞培养液中超氧化物歧化酶(SOD)活力、TBA法测定丙二醛(MDA)含量、硝酸还原酶法测定一氧化氮(NO)含量,UPLC/Q-TOF-MS法分析侧柏炭各溶剂提取物中黄酮类成分的差异。结果:与模型组比较,正丁醇提取物(100 mg·L-1)及乙酸乙酯提取物(100,50 mg·L-1)可显著提高细胞活力(P<0.05),显著降低MDA,NO含量,提高SOD活性(P<0.05)。4个溶剂提取物中,乙酸乙酯提取物中的黄酮总量含量最高,水提取物中黄酮总量含量最低,正丁醇提取物中的黄酮总量与石油醚提取物相当,只是其所含的槲皮苷、杨梅苷含量仅次于乙酸乙酯提取物中的槲皮苷、杨梅苷含量。结论:侧柏炭乙酸乙酯提取物可显著拮抗LPS对HUVECs的损伤,为侧柏炭保护血管内皮细胞的最有效提取物,其所含的槲皮苷、杨梅苷或者多种黄酮类成分可能为其保护血管内皮细胞作用的活性物质,其机制可能与其能减少NO的产生,抑制细胞内脂质过氧化有关。

Objective： To study the protective effect of different solvent extracts from Platycladi Cacumen Carbonisatum （PCC） on LPS-induced human umbilical vein endothelial cell damage, and discuss the effective extracts from PCC for protecting vascular endo- thehal cells and their possible active substances. Method： HUVECs were cultured in vitro; And LPS was adopted to establish the hu- man umbilical vein endothelial cell damage model. MTT colorimetric method was used to determine cell activity; Xanthine oxidase method was adopted to detect the activity of superoxide dismutases （SOD） in the cell culture fluid ; The TBA method was adopted to determine the content of malondialdehyde （MDA） ; The nitrate reductase method was used to detect the content of nitric oxide （NO） ; And UPLC/Q-TOF-MS was used to analyze the difference in flavonoids components among different solvent extracts from PCC. Result： Compared with the model group, N-butanol extract （ 100 mg· L-1 ） and ethylacetate extract （ 100,50 mg·L-1 ） could significantly enhance the cell activity （ P 〈 0. 05 ）, significantly reduce MDA and NO content, and increase SOD activity （ P 〈 0.05 ）. Among the four solvent extracts, the content of total flavonids were the highest in ethyl acetate extract, the lowest in water extract and equivalent in N-butanol and petroleum benzene extract. In terms of the contents of quercitrin and myricitrin, N-butanol extract were second only to ethyl acetate extract. Conclusion： Ethylacetate extract from PCC has a notable antagonistic effect in the damage induced by LPS to HUVECs, and thus is the most effective extract from PCC in protecting vascular endothelial cells. Quercitrin, myricitrin or multiple fla- vonoids that it contains may be their active substances for protecting vascular endothelial cells. Its mechanism may be related to the decrease in the production of NO and the inhibition of lipid peroxidation in ceils.