Effect of baicalin on liver fatty acid binding protein in oxidative stress model in vitro / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of baicalin on liver fatty acid binding protein in oxidative stress model in vitro.</p><p><b>METHODS</b>(1) Cellular oxidative stress in vitro was induced by incubating cells with 400μmol/L hydrogen peroxide (H₂O₂) for 20 minutes at 37 degrees C in the dark. After Chang liver cell line was treated with different dose of baicalin for 24, 48 and 72 hours. MTT assay was employed to detect cell viability, and then the hydrogen peroxide (TC50) of the different dose of baicalin was calculated. (2) Based on MTT assay, cells were treated with three different doses of baicalin (25, 50, 100 μmol/L) for 24 and 48 hours before being exposed to 400 μmol/L H₂O₂ for 20 minutes at 37 degrees C. Then, reactive oxygen species (ROS) assay and activity assays of superoxide dismutase (SOD) and reduced glutathione hormone (GSH) were evaluated. (3) Realtime PCR and Western blotting were applied to explore the influence of baicalin on the expression level of L-FABP. (4) One-way ANOVA was used for results statistical analysis.</p><p><b>RESULT</b>(1) MTT assay showed baicalin treatment at 25, 50, 100 μmol/L for 24 and 48 hours was feasible (83.60% ± 3.47%, 72.36% ± 2.18%, 70.16% ± 2.04% for 24 hours; 84.93% ± 3.11%, 76.16% ± 2.45%, 72.72% ± 2.31% for 48 hours, P > 0.05, F = 386.24, 475.92 respectively). Meanwhile, we found by the linear regression model that the median toxic concentration of baicalin for 48 hours was 170.6 μmol/L, and the median toxic concentration of baicalin for 24 hours was 153.2 μmol/L. (2) ROS assay showed dichlorofluorescin in all baicalin-treated cells after stress was significantly reduced (37.0 ± 3.30, 22.90 ± 3.84, 29.60 ± 2.52 for 24 hours respectively, P < 0.05, F = 70.06; 35.77 ± 2.35, 21.80 ± 3.10, 23.87 ± 1.98 for 48 hours respectively, P < 0.05, F = 110.92) as compared with the H₂O₂-treated cells. Moreover, 50 μmol/L baicalin treatment for 48 hours was the optimal condition against ROS generation (21.80 ± 3.10, P < 0.01, F = 110.92). Furthermore, the activities of intracellular SOD and GSH was increased significantly (51.53 ± 1.91 μg/mg for SOD, P < 0.05, F = 93.81; 49.85 ± 1.45 U/mg for GSH, P < 0.05, F = 92.51). (3) Although realtime PCR analysis indicated 50 μmol/L baicalin treatment for 48 hours could have no changes of the level of L-FABP expression under the oxidative stress condition, western blotting analysis indicated 50 μmol/L baicalin treatment for 48 hours could increase up to about 80% for the level of L-FABP expression.</p><p><b>CONCLUSION</b>Baicalin was suggested to be able to enhance both L-FABP expression and activity of intracellular SOD and GSH, and therefore protected hepatocytes from oxidative stress.</p>

The expression and significance of osteopontin in the development of nonalcoholic fatty liver fibrosis in rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the changes of osteopontin (OPN) in the liver tissues during nonalcoholic fatty liver fibrosis in rats and to explore the effect of OPN in the development of nonalcoholic fatty liver fibrosis.</p><p><b>METHODS</b>Fifty-six male Wistar rats were randomly divided into a control group (8 rats) and a high-fat diet group. The high-fat diet group was divided into 6 subgroups (8 rats in each subgroup) with high-fat feedings for 4, 8, 12, 16, 20 or 24 weeks. Conventional histochemical, HE, Masson-trichrome and immunohistochemical staining for alpha-smooth muscle actin (a-SMA) were performed with the liver histological preparations. The expression of OPN was detected with reverse transcription and polymerase chain reactions and Western blot.</p><p><b>RESULTS</b>Levels of OPN in liver tissues in rat nonalcoholic fatty liver fibrosis induced by high-fat diet were significantly increased over those in the control group (F=7.15, P less than 0.01). OPN expressions were closely correlated with a-SMA and nonalcoholic fatty liver fibrosis, and correlation coefficients of the two groups were 0.94 and 0.82, and both P values were less than 0.01.</p><p><b>CONCLUSION</b>Expression of OPN increases dramatically in the livers during the development of nonalcoholic fatty liver fibrosis, and OPN may play an important role in this event.</p>

The significance and effects of liver X receptor alpha in nonalcoholic fatty liver disease in rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To explore liver X receptor alpha (LXR alpha) gene changes and their significance in nonalcoholic fatty liver disease (NAFLD) in rats.</p><p><b>METHODS</b>A rat model of nonalcoholic fatty liver disease was produced with a fatty diet regime (feeding group, FG). Rats fed with normal diet served as controls (CG). The mRNA and protein expressions of LXR alpha in liver tissues were detected by reverse transcription and polymerase chain reaction (RT-PCR) and Western blot.</p><p><b>RESULTS</b>The concentration of free fatty acid (FFA) in the sera of GF rats started to increase to 0.33 mmol/L after 4 weeks of fat diet feeding, while the FFA of the CG was just 0.24+/-0.03 mmol/L, and the difference was significant (P<0.05). The concentration of ALT and AST in sera of the FG rats started to increase to 75.8 U/L and 138.9 U/L at the 8th week, much higher than those of the CG (P<0.01), and at the 12th week they increased further (P<0.01). Meanwhile, the mRNA and protein expressions of LXR alpha at the 2nd week was significantly increased to 0.62 (P>0.01) and its peak was reached at the 12th week (P<0.01). There was a significant positive correlation between the expression of LXR alpha and the degree of NAFLD.</p><p><b>CONCLUSION</b>The changes of LXR alpha gene are closely related to the development of nonalcoholic fatty liver disease.</p>