Exploration of Perturbed Liver Fibrosis-Related Factors and Collagen Type I in Animal Model of Non-Alcoholic Fatty Liver Disease.

To determine their involvement in the onset of the disease, we investigated the changing levels of liver fibrosis-related proteins, namely, type-I collagen, α-smooth muscle actin (α-SMA), and transforming growth factor ß1 and ß3 (TGF-ß1, ß3). The four groups of Sprague-Dawley (SD) rats were involved in the study, namely, (i) normal control group, (ii) high-fat diet group (HFD), (iii) carbon tetrachloride (CCl4) group, and (iv) NAFLD group (animal model) which were chosen at random. The NAFLD model received HFD combined with subcutaneous injection of small doses of CCl4. Histopathological examination confirmed extent of liver fibrosis, while other immunological and molecular methods were used to evaluate expression and distribution of α-SMA, type I collagen TGF-ß1 and TGF-ß3, at both m-RNA and protein levels. In contrast to the normal control group, the NAFLD group showed moderately elevated expressions of TGF-ß1, α-SMA, and type I collagen, which was proportional on temporal scale of NAFLD persistence in the model (P < 0.05). In the early phage of NAFLD, enhancement in the mRNA transcripts and, henceforth, protein expression of TGF-ß3 was observed. However, these were found to be downregulated in case of liver fibrosis (P < 0.05). This NAFLD rat model shows the histopathologic changes of human NAFLD and is suitable for the study of NAFLD pathogenesis. These findings suggest that type I collagen and the liver fibrosis-related factors TGF- ß1, TGF- ß3, and α-SMA may be significant contributors to NAFLD. Although NAFLD model is previously demonstrated by other researchers, our study is novel in terms of exploration of involvement of fibrosis-related factors and in particular aforementioned proteins at the early stage of NAFLD vis-à-vis dynamics of type-I collagen distribution.

The Value of Color Doppler Ultrasound and CT Combined with Serum AFP Examination in the Diagnosis of Hepatocellular Carcinoma.

Objective: To evaluate the value of the combination of color Doppler ultrasound, computed tomography (CT), and serum tumor marker alpha-fetoprotein (AFP) examination in the diagnosis of hepatocellular carcinoma (HCC). Methods: 98 patients with HCC (malignant tumor group) and 50 liver lesion patients (benign control group), were selected for the study, and retrospective statistical methods were used to evaluate the diagnostic values of the three examinations on hepatocellular carcinoma. Results: (1) When comparing color Doppler ultrasound blood flow parameters, the hepatic artery diameter, peak flow velocity, minimum flow velocity, and resistance index (RI) of hepatocellular carcinoma were significantly higher than those of the benign control group (P < 0.05), while the portal vein flow velocity was significantly lower than that of the control group (P < 0.05). (2) Enhanced CT imaging of hepatocellular carcinoma lesions showed mostly outflow-type enhancement changes, with high- or slightly high-density shadowing and uneven enhancement in the arterial phase, relatively low density and withdrawal of enhancement in the portal vein phase and delayed phase. (3) The serum AFP level of hepatocellular carcinoma patients was significantly higher than that of the benign control group (P < 0.01). (4) The sensitivity of color Doppler ultrasound, CT, and serum AFP alone for the diagnosis of HCC was 79.59%, 85.71%, and 66.33%, and the accuracy was 83.78%, 87.16%, and 74.32%, respectively, while the combination of the three tests could significantly increase the sensitivity to 96.94% and the accuracy to 93.92%, compared with each individual test (P < 0.01). Conclusion: Color Doppler ultrasound and CT combined with serum AFP examination could significantly improve the sensitivity and accuracy of hepatocellular carcinoma diagnosis, reduce misdiagnosis, and facilitate early diagnosis and clinical early intervention.

[Effects of cAMP-response element binding protein-1 (CREB-1) on transforming growth factor-b3 (TGFb3) mRNA expression and promoter activity in hepatic stellate cells].

OBJECTIVE: To investigate the effects of cAMP-response element binding protein-1 (CREB-1) on transforming growth factor-b3 (TGF b3) mRNA expression and promoter activity in hepatic stellate cells (HSCs). METHODS: Freshly isolated HSCs from rats were divided into six groups: CREB-1 expression plasmid transfected group (C), siRNA-CREB-1 plasmid transfected group (S), negative control group (N), forskolin treated group (F), H-89 treated group (H), and blank group (B). Rats in each group were further sub-divided according to whether (+) or not (-) they were exposed to exogenous TGF b3. TGF b3 mRNA expression was measured by real time quantitative PCR. HSCs of the C, S, N, F, H and B groups were transfected with the TGF b3 promoter luciferase reporter plasmid (PGL3-TGF b3-P; W group), the TGF b3 promoter luciferase reporter plasmid with CRE mutation (PGL3-basic-TGF b3P-mCRE; M group) and the renilla luciferase control plasmid (pRL-SV40; control group). TGF b3 promoter activity was assessed by luciferase reporter assays. RESULTS: Compared to N(-), the TGF b3 mRNA expression was reduced to 0.69+/-0.15 in S(-) (P less than 0.05) and increased to 4.68+/-2.76 in C(-) (P more than 0.05). Compared to B(-), the TGF b3 mRNA expression was reduced to 0.57+/-0.08 in H(-) (P less than 0.05). The differences between N(+) and N(-), S(+) and S(-), B(+) and B(-), and H(+) and H(-) were all significant (P less than 0.05). The values of TGF b3 promoter activity in S(W), N(W), and C(W) were 0.062+/-0.013, 0.122+/-0.011, and 0.165+/-0.016 (P less than 0.05), but the changes of TGF b3 promoter activity in S(M), N(M), and C(M) were not significant (P more than 0.05). The values of TGF b3 promoter activity in H(W), B(W), and F(W) were 0.154+/-0.010, 0.188+/-0.016, and 0.276+/-0.031 (P less than 0.05), but the changes of TGF b3 promoter activity in H(M), B(M), and F(M) were not significant (P more than 0.05). CONCLUSION: Increased levels of CREB-1 mRNA or p-CREB-1 up-regulate the TGF b3 mRNA expression and promoter activity in rat HSCs. The CRE site in the TGF b3 promoter is critical for this effect, and the gene's activity becomes significantly decreased when the site is missing. Exogenous TGF b3 enhances expression of endogenous TGF b3 in rat HSCs.