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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400μg/mL、350μg/mL及125μg/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16.14μmol/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。
In this study, we screened from five extracts of medicinal and edible plants and Salvia miltiorrhiza active substances that have anti-inflammatory effects on high glucose-induced HBZY-1 cells. Cell proliferation was detected by MTT method. The contents of LDH, TNOS and iNOS were determined by microplate method. The content of NO was detected by nitrate reductase method. The content of ET-1 and the cells stained with hematoxylin and eosin Differences in cell morphology. The results showed that LBP, mulberry flavone and flavonoid extract of Ginkgo biloba flavonoids could inhibit the proliferation of HBZY-1 cells exposed to high glucose in a dose-dependent manner with concentrations of 400μg / mL, 350μg / mL and 125μg / mL. The above three extracts could inhibit the secretion of LDH, ET-1 and iNOS, and promote the expression of NO. The LDH production of flavonoids extract of Ginkgo biloba was 717.86 U / g prot, close to the positive control, the NO production was 16.14 μmol / L, which was superior to the positive control. The amount of ET-1 produced in the mulberry leaf flavonoid extract group was 26.42 pg / mL, which was better than the positive control and could protect and maintain good cell morphology. The three active substances obtained in this experiment have anti-inflammatory effects on high glucose-induced injury of HBZY-1 cells and can be used as raw materials for health food to improve the health status of patients with kidney disease.