Fig 1.
The flow chart of modeling control, NAFL, NASH and NAFL + T2DM rats.
The STZ (25 mg/kg in 0.1 M citrate buffer) and citrate buffer were given to rats through a single intraperitoneal injection. SD, standard diet; HFD, high fat diet; MCDD, methionine-choline-deficient diet; STZ, streptozotocin.
Fig 2.
The establishment of rat models in NAFL, NASH and NAFL + T2DM.
(A) Changes in body weight of rats during standard diet (control), high fat diet (NAFL and NAFL + T2DM) and methionine-choline-deficient diet (NASH) feeding. (B) Liver index (the ratio of liver weight to body weight) of the rats at the end of the experiment. Liver histological examination with H&E (x 200) (C) and Oil-Red-O staining (x 200) (D). Data are expressed as the mean ± SD (n = 10 per group). *P < 0.05 compared with the control group; §P < 0.05 compared with the NAFL group.
Table 1.
General and biochemical parameters of rats in different groups.
Fig 3.
Validation of selected genes in microarray data by qrt-PCR.
Hepatic mRNA levels of srebf1 (A), tbx21 (B), gimap4 (C), and s100a9 (D). The mRNA levels were normalized to those of 18S, and subsequently normalized to those of control group. *, P < 0.05 compared with the control group; §, P < 0.05 compared with the NAFL group.
Fig 4.
The levels of selected biomarkers in liver and serum.
(A) Hepatic S100A9, SREBF1, TBX21, and GIMAP4 protein levels in control, NAFL, NASH and NAFL + T2DM groups. (B) Serum levels of SREBF1, TBX21, GIMAP4, S100A9, and CK18 protein in the control, NAFL, NASH, NAFL+T2DM groups. *, P < 0.05 compared with the control group; §, P < 0.05 compared with the NAFL group.
Fig 5.
Linear regression analysis between serum S100A9 level and some clinical parameters, and its diagnostic value in definite NASH.
Correlation between serum S100A9 level and (A) NAS, steatosis, lobular inflammation, (B) ALT, AST, TBil, (C) FBG, HOMA-IR, TG and TC in rats of all groups. (D) The receiver operating characteristic (ROC) curve of S100A9 and CK18 for the definite NASH diagnosis in rats. Area under the ROC (AUROC) for S100A9 and CK18 is 0.947 and 0.931 respectively.