[关键词]
[摘要]
研究从五种药食同源植物及丹参的提取物中筛选出对高糖诱导HBZY-1细胞具有抗炎作用的活性物质。利用MTT法检测细胞增殖活性进行初步筛选,采用微板法检测LDH、TNOS和iNOS含量,硝酸还原酶法检测NO含量,ELISA法检测检测ET-1含量,苏木素伊红(HE)染色细胞并观察细胞形态学差异变化。结果表明:枸杞多糖、桑叶黄酮及银杏叶黄酮的提取物对高糖损伤的HBZY-1细胞增殖保护作用均呈现浓度依赖型,最佳作用浓度分别为400 μg/mL、350 μg/mL及125 μg/mL。以上三种提取物均能抑制细胞分泌LDH、ET-1、iNOS,促进NO表达,且银杏叶黄酮提取物组LDH生成量为717.86 U/g prot,接近阳性对照,NO生成量为16.14 μmol/L,优于阳性对照,桑叶黄酮提取物组ET-1生成量为26.42 pg/mL,优于阳性对照,皆能保护并维持良好的细胞形态。本试验得到的三种活性物质对高糖诱导HBZY-1细胞损伤具有抗炎作用,可以作为改善肾脏疾病患者健康状况的保健类食品原料。
[Key word]
[Abstract]
Active ingredients (from the extracts of five functional food plants and Salvia) with an anti-inflammatory effect on high-glucose-induced injury to HBZY-1 cells were screened here. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to measure cell proliferation in the initial screening, and lactate dehydrogenase (LDH), total nitric oxide synthase (TNOS), and inducible nitric oxide synthase (iNOS) were quantified by the microplate method. The concentrations of nitric oxide (NO) and endothelin (ET-1) were determined by the nitrate reductase method and enzyme-linked immunosorbent assay (ELISA), respectively. Hematoxylin and eosin (HE) staining was used for examining morphological changes in these cells. The results showed that the extracts of polysaccharides from Lycium barbarum (LBP), flavonoids from mulberry leaves, and flavonoids from Ginkgo biloba leaves had a protective effect against the high-glucose-induced proliferation of HBZY-1 cells in a concentration-dependent manner. The optimal concentrations of the three extracts were 400, 350, and 125 μg/mL, respectively. The three above-mentioned extracts inhibited the secretion of LDH, ET-1, and iNOS and promoted the production of NO. In addition, the amount of LDH yielded by the extract of Ginkgo biloba leaves was 717.86 U/(g prot), which was close to that of the positive control, and the production of NO was 16.14 μmol/L, which was higher than that of the positive control. The amount of ET-1 yielded by the extract of mulberry leaves was 26.42 pg/mL, which was higher than that of the positive control. All the three extracts protected and maintained cell morphology well. All the three active ingredients in this experiment were found to have an anti-inflammatory effect on the HBZY-1 cell injury induced by a high concentration of glucose and can be used as functional food ingredients for improving the health of patients with renal disease.
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[基金项目]
国家自然科学基金项目(31401487);山东省农业科学院科技创新重点项目(2014CXZ03)