[關(guān)鍵詞]
[摘要]
目的 基于細(xì)胞代謝組學(xué)技術(shù),考察商陸皂苷甲(EsA)誘導(dǎo)人腎小管上皮HKC細(xì)胞損傷前后對(duì)細(xì)胞代謝產(chǎn)物的影響,尋找潛在的生物標(biāo)志物。方法 MTT法檢測EsA 0(對(duì)照組)、0.125、0.250、0.500、0.750、1.000 mmol/L對(duì)HKC細(xì)胞活力的影響;試劑盒法檢測EsA 0(對(duì)照組)、0.25、0.75 mmol/L對(duì)HKC細(xì)胞上清LDH含量的影響,明確EsA腎細(xì)胞毒性;利用UPLC-Q/TOF-MS分析技術(shù),將EsA 0(對(duì)照組)、0.25、0.75 mmol/L作用24 h的HKC細(xì)胞進(jìn)行代謝組學(xué)分析,采用Progenesis QI軟件進(jìn)行數(shù)據(jù)處理,將得到的數(shù)據(jù)導(dǎo)入SIMCA-P12.0 software進(jìn)行多元統(tǒng)計(jì)分析,利用主成分分析(PCA)剔除離群值,進(jìn)行正交偏最小二乘分析(OPLS-DA)得到變量權(quán)重值(VIP),選取VIP>1的差異代謝物,進(jìn)行檢驗(yàn),獲得具有統(tǒng)計(jì)學(xué)意義的生物標(biāo)志物。通過HMDB、KEGG、METLIN等代謝物數(shù)據(jù)庫對(duì)生物標(biāo)記物進(jìn)行鑒定,利用MetPA平臺(tái)對(duì)進(jìn)行代謝通路分析。結(jié)果 與對(duì)照組比較,EsA在大于0.25 mmol/L時(shí)對(duì)HKC細(xì)胞活力有顯著的抑制作用(P<0.05、0.01);細(xì)胞上清液中LDH含量顯著上升(P<0.05)。通過代謝組學(xué)技術(shù)和相關(guān)數(shù)據(jù)庫分析篩選出了15個(gè)生物標(biāo)志物,主要涉及甘油磷脂代謝、嘧啶代謝、鞘脂代謝、氨基酸代謝和能量代謝途徑。結(jié)論 EsA可誘導(dǎo)腎細(xì)胞損傷,可能與氧化應(yīng)激損傷,氨基酸代謝失調(diào),能量代謝異常以及脂質(zhì)代謝紊亂相關(guān)。
[Key word]
[Abstract]
Objective To investigate the effects of Esculentoside A (EsA) on cellular metabolites before and after injury of human renal tubular epithelial HKC cells, and to find potential biomarkers based on cell metabolomics techniques. Methods MTT method was used to detect the effect of ESA 0 (control group), 0.125, 0.250, 0.500, 0.750 and 1.000 mmol/L on the activity of HKC cells; Kit method was used to detect the effect of ESA 0 (control group), 0.25 and 0.75 mmol/L on the content of LDH in the supernatant of HKC cells, so as to determine the nephrotoxicity of EsA; By using UPLC-Q/TOF-MS analysis technology, the HKC cells treated with EsA 0 (control group), 0.25 and 0.75 mmol/L for 24 hours were analyzed by metabonomics. The data were processed by Progenesis QI software and imported into SIMCA-P12.0 Software carries on multivariate statistical analysis, uses PCA to eliminate outliers, carries on OPLS-DA to obtain variable weight value (VIP), selects VIP > 1 differential metabolite, carries on the test, obtains the biomarker with statistical significance. Biomarkers were identified by HMDB, KEGG, metlin and other metabolite databases, and metabolic pathways were analyzed by metpa platform. Results After stimulation with EsA, HKC cells showed a decrease in cell viability (P < 0.05) and up-regulation of LDH in cell supernatant (P < 0.05), 15 biomarkers were screened by metabolomics technology and related database analysis, mainly related to glycerophospholipid metabolism, pyrimidine metabolism, sphingolipid metabolism, amino acid metabolism and energy metabolism pathway. Conclusion EsA can induce renal cell injury, may be associated with oxidative stress damage, amino acid metabolism disorders, abnormal energy metabolism, and lipid metabolism disorders.
[中圖分類號(hào)]
R965
[基金項(xiàng)目]