目的 從抑制早期生長反應因子-1（Egr-1）介導的白細胞介素8（IL-8）表達的角度探討強肝膠囊對大鼠非酒精性脂肪肝的改善作用。方法 Wistar大鼠隨機分為對照組、模型組和強肝膠囊組，每組各10只。模型組和強肝膠囊組喂食高脂飼料，建立高脂血癥非酒精性脂肪肝模型，造模同時強肝膠囊組大鼠ig強肝膠囊內容物0.53 mg/g，給藥體積為1 mL/100 g體質量，模型組ig等量生理鹽水。每周給藥6 d，持續(xù)8周。治療結束取動物肝臟，氧化酶法測定肝組織中三酰甘油（TG）、總膽固醇（TC）水平；實時熒光定量PCR（RT-qPCR）法檢測Egr-1和IL-8 mRNA表達；Western blotting法測定Egr-1蛋白表達；ELISA法測定肝組織中IL-8水平。將大鼠肝臟HepG2細胞分別進行0.25 mmol/L油酸處理、轉染Egr-1 siRNAs并油酸處理、轉染pcDNA3.1-Egr-1陽性表達質粒并油酸處理后，RT-qPCR法檢測細胞中Egr-1和IL-8 mRNA表達，WB法檢測Egr-1蛋白表達，ELISA法檢測培養(yǎng)液上清中IL-8水平。結果 經過8周治療，強肝膠囊組TG和、TC水平明顯下降（P<0.01）；Egr-1 mRNA、Egr-1蛋白表達和IL-8 mRNA、IL-8水平均明顯下降（P<0.01）。Egr-1 siRNA#1、siRNA#2能明顯降低Egr-1表達水平，采用siRNA干擾Egr-1基因，并用油酸處理HepG2細胞后，細胞內和培養(yǎng)液上清中IL-8水平較對照細胞顯著降低（P<0.01）。轉染Egr-1陽性表達質粒導致Egr-1表達量上升（P<0.01），IL-8表達也同步升高（P<0.01）。結論 Egr-1可正向調節(jié)IL-8表達。強肝膠囊通過抑制Egr-1介導的IL-8表達，達到改善非酒精性脂肪肝的作用。

Objective To discuss the improvement of Qianggan Capsules on non-alcoholic fatty liver in rats based on regulation of Egr-1 on IL-8 expression. Methods Wistar rats were randomly divided into the control, model, and Qianggan Capsules groups, and each group had 10 rats. Rats in model and Qianggan Capsules groups were fed high-fat diet to establish hyperlipidemic nonalcoholic fatty liver model. At the same time, the rats in Qianggan Capsules group were ig administered with Qianggan Capsules 0.53 mg/g, and the volume was 1 mL/100 g body weight. And rats in model group were given equivalent saline. Rats were treated 6 d weekly for 8 weeks. After treatment, liver tissues were separated. The levels of three glycerol oxidase (TG) and total cholesterol (TC) level in liver tissue were measured by oxidase method. Egr-1 and IL-8 expression of mRNA were detected by real-time fluorescence quantitative PCR (RT-qPCR) method, Egr-1 protein expression was detected by Western blotting (WB) method, and IL-8 levels were detected by ELISA method. The HepG2 cells were treated with 0.25 mmol/L oleic acid, transfected with Egr-1 siRNAs and then treated with oleic acid, or transfected with pcDNA3.1-Egr-1 and then treated with oleic acid. The expression of Egr-1 and IL-8 mRNA in the cells was detected by RT-qPCR method, the expression of Egr-1 protein was detected by Western blotting method, and IL-8 level in the supernatant of the culture liquid was detected by ELISA method. Results After treatment for 8 weeks, the levels of TG and TC were significantly decreased in Qianggan Capsules group (P < 0.01), and the expression of Egr-1 mRNA and Egr-1 protein, IL-8 mRNA and IL-8 levels were significantly decreased (P < 0.01). Egr-1 siRNA#1 and siRNA#2 could significantly decrease the levels of Egr-1 expression. After siRNA interference with Egr-1 gene and HepG2 treated with oleic acid, IL-8 levels in cells and culture supernatants were significantly lower than those in control cells (P < 0.01). The expression of Egr-1 was increased by transfection of Egr-1 positive expression plasmid (P < 0.01), and IL-8 expression also increased (P < 0.01). Conclusion Egr-1 can positively regulate the expression of IL-8. Qianggan Capsules can improve the effect of nonalcoholic fatty liver by inhibiting the expression of Egr-1 mediated IL-8.

天津市衛(wèi)生和計劃生育委員會中醫(yī)中西醫(yī)結合科研課題（2015107）