Effects of Baicalin on High Fat-Induced Lipid Accumulation and SIRT1 Related Factors Expression of HepG2 Cells in vitro
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摘要: 目的 研究黄芩苷(Baicalin, BA)对游离脂肪酸(FFA)诱导HepG2细胞脂肪沉积的影响及机制研究。方法 选用0.75 mmol/L的FFA体外诱导HepG2细胞24 h，建立体外脂肪沉积模型。将细胞分为正常组、模型组、黄芩苷低、中、高剂量组。利用油红O染色及GPO-PAP酶法检测各组细胞内甘油三酯(TG)含量的变化，ELISA法检测各组细胞上清液炎症因子TNF-α、IL-6分泌量。Western Blot检测各组细胞内沉默信息调节因子1(SIRT1)、碳水化合物应答元件结合蛋白(ChREBP)、胆固醇调节元件结合蛋白-1c(SREBP-1c)以及脂肪酸合成酶(FAS)的蛋白含量。结果 FFA诱导的HepG2细胞内脂质积聚明显，细胞内TG含量显著上升；药物干预后，3个药物组细胞内脂滴较模型组均有减少，且呈药物浓度依赖性；细胞内TG含量呈下降趋势，其中中剂量和高剂量药物组与模型组相比，差异具有统计学意义(P < 0.05)；Western Blot检测结果显示，与空白组相比，模型组SIRT1蛋白表达显著降低，SREBP-1c以及FAS的蛋白表达显著上升，与模型组相比，中、高剂量黄芩苷组细胞SIRT1蛋白表达显著上升，SREBP-1c、ChREBP以及FAS蛋白表达显著下降(P < 0.05)。结论 黄芩苷可以改善FFA诱导的HepG2细胞的脂肪沉积，减低细胞内TG的含量，其机制可能与调节SIRT1/SREBP-1c通路相关。Abstract: OBJECTIVE To study the effects and mechanism of baicalin on high fat-induced lipid accumulation in HepG2 cells.METHODS HepG2 cells were induced in vitro with 0.75 mmol/L free fatty acid for 24 h to establish an in vitro fat deposition model. The cells were then divided into the following groups; normal group, model group, 50 μmol/L baicalin group, 100 μmol/L baicalin group and 200 μmol/L baicalin group. Oil Red O staining and GPO-PAP enzymatic method were used to detect the changes of TG content in each group of cells, and enzyme-linked immunosorbent assay (ELISA) was used to detect inflammatory factors in the supernatant of each group: TNF-α, IL-6. Western Blot was used to detect the following protein content: SIRT1, ChREBP, SREBP1c, FAS Protein.RESULTS Free fatty acid induced lipid accumulation in HepG2 cells increased, the concentration of TG in cells increased significantly; after the drug intervention, the intracellular lipid droplets in the three drug groups were reduced in comparison to the model group, therefore, showing drug-concentration-dependent results; the TG concentration showed a downward trend in the drug-groups, in which, the differences between the 100 μmol/L and 200 μmol/L drug group and the model group were statistically significant (P < 0.05). Western blot detection showed that compared with the normal group, the expression of SIRT1 protein in the model group was significantly reduced, and the protein expressions of SREBP-1c and FAS increased significantly; the SIRT1 protein expressions of the cells in the 100 μmol/L baicalin group and 200 μmol/L baicalin group increased significantly, while SREBP-1c, ChREBP and FAS protein expression decreased significantly (P < 0.05).CONCLUSION Baicalin can reduce the lipid accumulation of HepG2 cells induced by free fatty acids and reduce the TG content in the cells. The mechanism may be related to the SIRT1/SREBP-1c pathway.
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Key words:
- lipid accumulation /
- baicalin /
- SIRT1 /
- SREBP-1c
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图 2 不同浓度FFA对HepG2细胞脂质积累的影响(×400)
注：A.空白对照；B.0.25 mmol/L FFA组；C.0.5 mmol/L FFA组；D.0.75 mmol/L FFA组；E.1 mmol/L FFA组；F.1.25 mmol/L FFA组
图 6 黄芩苷对HepG2细胞内SIRT1、SREBP1c、ChREBP、FAS蛋白表达的影响
注：A.正常组；B.模型组；C.低剂量黄芩苷；D.中剂量黄芩苷；E.高剂量黄芩苷。与空白组相比，△P < 0.05；与模型组相比，*P < 0.05, **P < 0.01。x±s，n=3。
表 1 不同浓度FFA诱导HepG2细胞生存率的比较(x±s, n=3)
FFA浓度/(mmol·L-1) 细胞生存率/% 0 109.26±8.58 0.25 99.62±5.31 0.50 92.19±8.98* 0.75 80.13±12.11** 1.00 70.08±10.40** 1.25 59.92±3.49** 注：与空白组相比，*P < 0.05, **P < 0.01。 
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表 2 各组细胞上清液炎症因子IL-6、TNF-α水平的比较(x±s，pg·mL-1, n=3)
组别 IL-6 TNF-α 正常组 91.76±9.12 79.49±17.52 模型组 108.40±11.10 103.83±13.38 低剂量组(50 μmol/L) 103.35±3.48 101.04±10.58 中剂量组(100 μmol/L) 100.69±14.28 100.65±18.85 高剂量组(200 μmol/L) 97.47±5.56 78.08±15.83
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[1] ROEB E, STEFFEN HM, BANTEL H, et al. S2k Guideline non-alcoholic fatty liver disease[J]. Z Gastroenterol, 2015, 53(7): 668-723. doi: 10.1055/s-0035-1553193 [2] ARAB JP, ARRESE M, TRAUNER M. Recent insights into the pathogenesis of nonalcoholic fatty liver disease[J]. Annu Rev Pathol, 2018, 13: 321-350. doi: 10.1146/annurev-pathol-020117-043617 [3] FRIEDMAN SL, NEUSCHWANDER-TETRI BA, RINELLA M, et al. Mechanisms of NAFLD development and therapeutic strategies[J]. Nat Med, 2018, 24(7): 908-922. doi: 10.1038/s41591-018-0104-9 [4] YAO H, TAO X, XU L, et al. Dioscin alleviates non-alcoholic fatty liver disease through adjusting lipid metabolism via SIRT1/AMPK signaling pathway[J]. Pharmacol Res, 2018, 131: 51-60. doi: 10.1016/j.phrs.2018.03.017 [5] GUO WW, WANG X, CHEN XQ, et al. Flavonones from Penthorum chinense ameliorate hepatic steatosis by activating the SIRT1/AMPK pathway in HepG2 cells[J]. Int J Mol Sci, 2018, 19(9): 2555. doi: 10.3390/ijms19092555 [6] CHANG YH, CHEN YL, HUANG WC, et al. Fucoxanthin attenuates fatty acid-induced lipid accumulation in FL83B hepatocytes through regulated Sirt1/AMPK signaling pathway[J]. Biochem Biophys Res Commun, 2018, 495(1): 197-203. doi: 10.1016/j.bbrc.2017.11.022 [7] DAI J, LIANG K, ZHAO S, et al. Chemoproteomics reveals baicalin activates hepatic CPT1 to ameliorate diet-induced obesity and hepatic steatosis[J]. PNAS, 2018, 115(26): E5896-E5905. doi: 10.1073/pnas.1801745115 [8] SHEN KZ, FENG XW, PAN H, et al. Baicalin ameliorates experimental liver cholestasis in mice by modulation of oxidative stress, inflammation, and NRF2 transcription factor[J]. Oxid Med Cell Longev, 2017, 2017: 6169128. [9] LIU J, YUAN YL, GONG X, et al. Baicalin and its nanoliposomes ameliorates nonalcoholic fatty liver disease via suppression of TLR4 signaling cascade in mice[J]. Int Immunopharmacol, 2020, 80: 106208. doi: 10.1016/j.intimp.2020.106208 [10] MARUYAMA H, KOBAYASHI K, KIYONO S, et al. Free fatty acid-based low-impedance liver image: A characteristic appearance in nonalcoholic steatohepatitis (NASH)[J]. Eur Radiol Exp, 2020, 4(1): 3. doi: 10.1186/s41747-019-0137-y [11] BUZZETTI E, PINZANI M, TSOCHATZIS EA. The multiple-hit pathogenesis of non-alcoholic fatty liver disease (NAFLD)[J]. Metabolism, 2016, 65(8): 1038-1048. doi: 10.1016/j.metabol.2015.12.012 [12] DING RB, BAO JL, DENG CX. Emerging roles of SIRT1 in fatty liver diseases[J]. Int J Biol Sci, 2017, 13(7): 852-867. doi: 10.7150/ijbs.19370 [13] LEE HS, LIM SM, JUNG JI, et al. Gynostemma pentaphyllum extract ameliorates high-fat diet-induced obesity in C57BL/6N mice by upregulating SIRT1[J]. Nutrients, 2019, 11(10): E2475. doi: 10.3390/nu11102475 [14] LUO X, HE ZY, SUN XM, et al. DHA protects against hepatic steatosis by activating Sirt1 in a high fat diet-induced nonalcoholic fatty liver disease mouse model[J]. Diab Metab Syndr Obes, 2020, 13: 185-196. doi: 10.2147/DMSO.S232279 [15] PONUGOTI B, KIM DH, XIAO Z, et al. SIRT1 deacetylates and inhibits SREBP-1C activity in regulation of hepatic lipid metabolism[J]. J Biol Chem, 2010, 285(44): 33959-33970. doi: 10.1074/jbc.M110.122978 -
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