p-JAK1/p-STAT3通路在蓝莓益生菌血清干预肝细胞脂肪变性中的作用

【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（p-JAK1）/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24 h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、p-JAK1、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。

英文摘要:

【Abstract】 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- and}_medium-doseserum groups were found (P >0.05). Conclusion The blueberry probiotics could antagonize the NAFLD via p-JAK1/p-STAT3 signaling way.

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