图1 各组小鼠的血糖、甘油三酯和总胆固醇水平及及肝组织上清液ALT及AST表达水平

Fig.1 Levels of blood glucose, triglycerides and total cholesterol and expression levels of ALT and AST in the liver tissue of the mice in each group (n=6, Mean±SD). A: Levels of blood glucose, triglyceride and total cholesterol in different groups. B: Expression levels of ALT and AST in the liver tissue in different groups. *P<0.05 vs NC; #P<0.05 vs NAFLD.

图2 各组肝组织中病理改变及脂肪堆积程度

Fig.2 Pathological changes and fat accumulation in the liver tissue of the mice in each group (n=6, Mean±SD). A: HE staining. B: Oil red O staining.

图3 ELISA法检测 IL-6表达水平

Fig.3 Expression of IL-6 in the liver tissues of the mice in each group (n=6, Mean±SD). *P<0.05 vs NC; #P<0.05 vs NAFLD.

图4 Western blotting、Real-time qPCR、免疫组化染色在各组肝组织中的表达和分布

Fig.4 Protein levels of Sp1 and FN in the liver tissues of the mice in each group determined by Western blotting (n=6, Mean±SD). A, B: Western blotting for detecting protein expression levels of FN and SP1. C: RT-qPCR for detecting expressions of Sp1 and FN mRNA. D: Immunohistochemical staining for Sp1 and FN in the liver tissues of the mice in each group. *P<0.05 vs NC; #P<0.05 vs NAFLD.

图5 各组肝组织中Col-Ⅳ、E-ca和α-SMA蛋白表达水平

Fig.5 Expression levels of Col-IV, E-ca and α-SMA in the liver tissues of the mice in each group detected by Western blotting (n=6, Mean±SD). *P<0.05 vs NC; #P<0.05 vs NAFLD.

图6 CCK8实验中各分组细胞存活率对比图

Fig.6 Comparison of cell survival rate of each group in CCK8 experiment (n=3, Mean±SD). A: Screening the safe concentration range of XJ-60 acting on AML12 cells without FFA intervention, P>0.05 vs NC. B: The optimal therapeutic concentration of XJ-60 acting on AML12 cells under FFA intervention. **P<0.01 vs PO.

图7 油红O染色观察各组肝细胞脂滴积累情况

Fig.7 　　Accumulation of lipid droplets in the hepatocytesin each group. A: Pathological changes and lipidaccumulation in hepatocytes of different groupsobserved using Oil Red O staining. B: Quantitativeanalysis of lipid accumulation in the hepatocytes in eachgroup (n=3, Mean±SD), **P<0.01 vs NC; ##P<0.01 vs PO.

图8 XJ-60对AML12细胞中SP1/TGF-β/Smad3信号通路的影响

Fig.8 Effect of XJ-60 on expressions of SP1/TGF-β/Smad3 signaling pathway in FFA-induced AML12 cells (n=3, Mean±SD). A, B: Protein expression levels of SP1, TGF-β and p-Smad3 in AML12 cells in each group (*P<0.05, **P<0.01 vs PO+DMSO). C: mRNA expression levels of SP1, TGF-β and p-Smad3 in hepatocytes of each group (**P<0.01,***P<0.001 vs PO+DMSO). D: SP1 and p-Smad3 immunofluorescence staining of AML12 cells in each group (×1000).

图9 敲低SP1后各指标的表达变化

Fig.9 Changes in expressions of the protein markers in AML12 cells after SP1 knockdown (n=3, Mean±SD). A, B: Protein expression levels of FN, Col-IV, SP1, TGF‑β and p-Smad3 in AML12 cells (*P<0.05, **P<0.01 vs PO+Neg). C: mRNA expression levels of FN, Col-IV, SP1, TGF-β and p-Smad3 in AML12 cells in each group (*P<0.05, **P<0.01 vs PO+Neg).

图10 XJ-60对EMT及ECM的改善情况

Fig.10 Effect of XJ-60 treatment on expressions of EMT and ECM markers in AML12 cells (n=3, Mean±SD). A, B: Protein expression levels of FN, Col-IV, E-ca and α-SMA in AML12 cells in each group (*P<0.05, **P<0.01, ***P<0.001 vs PO+DMSO). C: mRNA expression levels of FN, Col-IV, E-ca and α-SMA in hepatocytes of each group (**P<0.01 vs PO+DMSO).

图11 CAA与XJ-60对AML12细胞纤维化的改善情况

Fig.11 Effects of CAA and XJ-60 for improving fibrosis in AML12 cells (n=3, Mean±SD). A: Protein bands of FN, Col-IV and α-SMA in AML12 cells in each group. B: Relative protein expression levels. **P<0.01, ****P<0.0001 vs PO+CAA.