以白毫银针水提取物为供试制剂，探讨其对LO2细胞脂质积累的影响及作用机理。通过油酸处理建立非酒精性脂肪肝细胞模型，分别检测白毫银针水提取物治疗组、预防组、辛伐他汀组、自然恢复组、模型组LO2细胞的甘油三脂(TG)和丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、脂质积累，并测定细胞内SREBP-1c、FAS和CPT1A mRNA表达变化。结果表明，1 mmol·L-1油酸处理24 h为建立非酒精性脂肪肝细胞模型的最理想条件，模型组细胞内TG和MDA含量高于正常组，但SOD活性低于正常组；水提取物处理各组别中，治疗组1 mg·mL-1白毫银针制剂处理的细胞MDA含量最低，脂肪颗粒最少，SREBP-1c和FAS相对表达量最低，但CPT1A相对表达量和SOD活性最高，为本实验非酒精性脂肪肝防治的最优处理。1 mg·mL-1白毫银针制剂能显著降低LO2细胞的脂质积累，其机制可能与抑制SREBP-1c及其下游的FAS基因表达，减少TG合成，增加CPT1A基因表达，加速TG分解等因素有关。
In this paper, the aqueous extract of silver needle tea was used as the experimental preparation for study of its effect and mechanism on the lipid accumulation in LO2 cells. The cell model of non-alcoholic fatty liver was established by the treatment of oleic acid, and the therapeutic group, preventive group, simvastatin group and natural recovery group were arranged and respectively applied to the above experimental preparations and detected for triglycerides (TG) and malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, lipid accumulation and SREBP-1c, FAS and CPT1A mRNA expressions in LO2 cells. In this investigation, 1 mmol·L-1 oleic acid treatment for 24 h was the optimal condition for establishment of non-alcoholic fatty liver cell model, the contents of TG and MDA in the model group were higher than that in the normal group, while the SOD activity was lower when compared with the normal group, and among the groups the treatment of 1 mg·mL-1 aqueous extract in therapeutic group showed the lowest MDA content, the lowest lipid accumulation，the lowest relative expression of SREBP-1c and FAS，but the highest SOD activity and the highest relative expression of CPT1A of LO2 cells, and was regarded as the optimal aqueous preparation for prevent and treat the non-alcoholic fatty liver. The lipid accumulation of LO2 cells was significantly reduced by the treatment of the 1 mg·mL-1 aqueous extract, which may be related to the inhibition of SREBP-1c and the successive FAS gene expression, the decrease of TG synthesis, the acceleration of CPT1A gene expression and decomposition of TG, etc.
[1] Day C P, James O F. Steatohepatitis: a tale of two "hits"?[J]. Gastroenterology, 1998,114(4): 842—845.
[2] Li X, Wang R, Zhou N, Wang X, Liu Q, Bai Y, Liu Z, Yang H, Zou J, Wang H, Shi T. Quercetin improves insulin resistance and hepatic lipid accumulation in vitro in a NAFLD cell model[J]. Biomed Rep, 2013,1(1): 71—76.
[3] Ben M D, Baratta F, Polimeni L, Angelico F. Non-alcoholic fatty liver disease and cardiovascular disease: epidemiological, clinical and pathophysiological evidences[J]. Internal & Emergency Medicine, 2012,7(3): 291—296.
[4] 莫志贤,邵红霞. 白藜芦醇苷体外对过氧化氢导致小鼠肝细胞损伤的保护作用[J]. 中国药理学通报, 2000,16(5):519.
[5] 万晓强,李孝生,曾庆贵,刘梁英. 大黄酸对TNF-α和TG诱导L-02肝细胞凋亡的影响[J]. 胃肠病学和肝病学杂志, 2006,15(4): 383—386.
[6] 李孝生,刘梁英,万晓强. 川芎嗪、大黄酸联用对转化生长因子β1和甘油三酯致肝细胞损伤的影响[C]. 哈尔滨: 中国中西医结合学会第十八次全国消化系统疾病学术会议, 2006.
[7] 叶红球. 枳椇子水提取液对乙醇体外诱导肝细胞株LO2细胞损伤的影响[D]. 长沙: 中南大学硕士学位论文, 2012.
[8] 黄玉影. 荔枝核有效部位群对实验性非酒精性脂肪肝的治疗作用和机制研究[D]. 广州: 广州中医药大学硕士学位论文, 2016.
[9] 岳文杰. 茶叶体外抗氧化效果研究[D]. 福州: 福建农林大学硕士学位论文, 2008.
[10] 王丽. 57份茶树种质资源的生化成分及抗氧化活性研究[D]. 福州: 福建农林大学硕士学位论文, 2012.
[11] Almajano M P, Carbó R, Jiménez J A L, Gordon M H. Antioxidant and antimicrobial activities of tea infusions[J]. Food Chemistry, 2008,108(1): 5—63.
[12] 杨伟丽,肖文军,邓克尼. 加工工艺对不同茶类主要生化成分的影响[J]. 湖南农业大学学报(自然科学版), 2001(5): 384—386.
[13] 黄赟. 福建白茶化学成分与感官品质研究初报[D]. 福州: 福建农林大学硕士学位论文, 2013.
[14] 张宁. 基于国际标准制定的白茶理化成分的研究[D]. 福州: 福建农林大学硕士学位论文, 2010.
[15] 周有良. 萎凋、造型工艺对白茶化学成分含量及品质的影响[D]. 福州: 福建农林大学硕士学位论文, 2010.
[16] 潘玉华,高树英,黄先洲,周寒松. 白茶萎凋温度对内含生化成分变化的影响[J]. 蚕桑茶叶通讯, 2013(3): 24—31, 33.
[17] 刘淑敏. 不同茶类浸提液及茶多酚的生物活性和机理研究[D]. 广州: 华南理工大学博士学位论文, 2016.
[18] Polimeni L, Del B M, Baratta F, Perri L, Albanese F, Pastori D, Violi F, Angelico F. Oxidative stress: New insights on the association of non-alcoholic fatty liver disease and atherosclerosis[J]. World Jounal of Hepatol, 2015,7(10): 1325—1336.
[19] Albano E. Free radical mechanisms in immune reactions associated with alcoholic liver disease.[J]. Free Radical Biology & Medicine, 2002,32(2): 110—114.
[20] Lu K H, Liu C T, Raghu R, Sheen L. Therapeutic Potential of Chinese Herbal Medicines in Alcoholic Liver Disease[J]. Journal of Traditional & Complementary Medicine, 2012,2(2): 115—122.
[21] Ding R B, Tian K, Huang L L, He C W, Jiang Y, Wang Y T, Wan J B. Herbal medicines for the prevention of alcoholic liver disease: A review[J]. Journal of Ethnopharmacology, 2012,144(3): 457—465.
[22] 叶寅. L-茶氨酸通过增强肝脏细胞中抗氧化物质活性抑制酒精性肝损伤的研究[D]. 南京: 南京大学硕士学位论文, 2011.
[23] Koteish A, Diehl A M. Animal models of steatosis.[J]. Seminars in Liver Disease, 2001,21(1): 89—104.
[24] Ferré P, Foufelle F. Hepatic steatosis: a role for de novo lipogenesis and the transcription factor SREBP-1c.[J]. Diabetes Obesity & Metabolism, 2010,12(S2): 83—92.
[25] Kohjima M, Enjoji M, Higuchi N, Kato M, Kotoh K, Yoshimoto T, Fujino T, Masayoshi Y, Yaoa R, Harada N, Takayanagi R, Nakamnta M. Re-evaluation of fatty acid metabolism-related gene expression in nonalcoholic fatty liver disease.[J]. International Journal of Molecular Medicine, 2007,20(3): 351—358.
[26] Jump D B, Tripathy S, Depner C M. Fatty acid-regulated transcription factors in the liver[J]. Annual Review of Nutrition, 2013,33(1): 249—269.
[27] Ahmed M H, Byrne C D. Modulation of sterol regulatory element binding proteins (SREBPs) as potential treatments for non-alcoholic fatty liver disease (NAFLD)[J]. Drug Discovery Today, 2007,12(17—18): 740—747.
闽公网安备 35020602000789号