摘要
目的建立体外肝细胞脂肪变性模型,探讨白介素-22(IL-22)调控的磷酸化酪氨酸蛋白激酶-1(pJAK1)/磷酸化信号转导激活转录因子-3(p-STAT3)信号通路在蓝莓益生菌血清改善肝细胞脂肪变性实验中的作用机制。方法制备蓝莓益生菌原液,并制备大鼠10%蓝莓益生菌低、中、高剂量血清及生理盐水血清。建立游离脂肪酸(FFA)诱导的正常肝细胞株L-02细胞脂肪变性模型,设立正常组,模型组,蓝莓益生菌低、中、高剂量血清组。定量检测细胞内三酰甘油(TG)含量;油红O染色检测细胞内的脂质沉积;RT-PCR和实时荧光定量PCR(qRT-PCR)检测各组IL-22、p-JAK1、p-STAT3、胆固醇调节元件蛋白-1c(SREBP-1c)的基因表达;免疫荧光及Western blot检测各组IL-22、p-JAK1、p-STAT3、SREBP-1c的蛋白表达。结果 FFA诱导刺激24h后,模型组细胞TG含量高于正常组(P<0.01),细胞内可见大量脂质沉积,且IL-22、p-JAK1、p-STAT3基因表达及蛋白表达均较正常组减弱(P均<0.01)、SREBP-1c基因及蛋白表达较正常组升高(P均<0.01);与模型组相比,蓝莓益生菌低、中、高剂量血清组的TG逐渐降低(P均<0.01),细胞内的脂肪沉积逐渐减轻,高剂量血清组IL-22、pJAK1、p-STAT3的基因表达和蛋白表达较模型组、低剂量血清组、中剂量血清组增强(P均<0.01),SREBP-1c的表达降低(P<0.01)。结论蓝莓益生菌通过对p-JAK1/p-STAT3信号通路的调节,参与拮抗肝细胞脂肪变性的作用。
Objective To explore the mechanism of interlukin-22 （IL-22）-mediated phosphor-Janus kinase-1（p-JAK1）/phosphor-signal transducer and activator of transcription 3 （p-STAT3） signaling way in the experiment of improving non-alcholic fatty liver disease （NAFLD） by blueberry probiotic serum.Methods The rat serums with low-, medium-, and high-dose of 10% blueberry probiotics, as well as saline were prepared. NAFLD model was built by inducing normal liver cell line L-02 with free fatty acid （FFA）.NAFLD model cells were cultured with saline serum （model group）, low-, medium-, and high-dose blueberry probiotics serums （low-, medium-, and high-dose serum groups） ,respectively .Normal liver cell group （normal group） was cultured with saline serum . Oil Red O staining was used to detect the lipid deposition in the cells; the intracellular level of triglyceride （TG） was quantitatively determined; the gene and protein expressions of IL-22, p-JAK1, p-STAT3, sterol-regulatory element binding protein-1c （SREBP-1c ） were detected by RT-PCR, Western blot and immunofluorescence methods. Results Twenty-four hours after modeling, a large amount of lipid deposition could be observed in model group. Compared with normal group, model group showed lower gene and protein expression levels of IL-22, p-JAK1 and p-STAT3 （P 〈0.01）, and higher SREBP-1c and TG levels （P 〈0.01）.Compared with model group, TG level and the lipid deposition in low-, medium-, and high-dose blueberry probiotics serum groups were gradually reduced. High-dose serum group showed higher gene and protein expression levels of IL-22, p-JAK1, p-STAT3 and lower SREBP-1c compared with the model, low-, and medium-dose serum groups （P 〈0.01）. No significant [CM（155.3mm]differences in gene and protein levels between low- andmedium-doseserum groups were found （P 〉0.05）. Conclusion The blueberry probiotics could antagonize the NAFLD via p-JAK1/p-STAT3 signaling way.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2017年第2期203-209,共7页
Journal of Sichuan University(Medical Sciences)
基金
贵州省科技厅社会攻关项目(No.黔科合SY[2010]3017号)
黔东南苗族侗族自治州林业局科研基金项目(No.ZLYJZFCG-2012-7)
贵州省卫计委科技项目(No.gzwjkj2016-1-02)资助
