Protective Effects Of Berberine From Chronic Liver Injury Induced By Carbon Tetrachloride

Liver injury can lead to hepatic fibrosis、cirrhosis、liver cancer and other diseases.It cando seriously harm to the health of people.Therefore，looking for the treatment of liver injuryhas become a arduous task.Modern medicine has discussed the mechanism of liver injury inmany ways and developed membrane protectant、anti-lipid oxidase agent、 anti-immuneresponse agent to protect liver.But the effect of these drugs was not apparent and their toxicand side-effect limit the clinical application.So It,s really important to develop efficient、lowtoxicity hepatoprotective drugs.Studies show that reactive oxygen species is closely related to the occurrence of liverinjury.On One hand,ROS can activate hepatic stellate cells and promote the synthesis ofcollagen in HSCs.Collagen is the main component of ECM,so ROS can promote thedeposition of ECM, which can urge the occurrence of hepatic fibrosis.On the otherhand,ROS can phosphorylate Akt, which will result in the synthesis of translation initiationcomplex.As a result,the production of collage increased. TGF-β1not only can increase thesynthesis of collagen,but also can inhibit it,s degradation.It can also activate HSCs andNOX,for NOX is the main sourse of ROS, thereby TGF-β1can increase the release ofROS.Thus,ROS,NOX, TGF-β1and Akt have played a important role in the development ofliver injury. It can be a crucial target to inhibit the producion or activation of these factors.AMPK is a kind of protein kinase,which regulates energy and metabolic balance inliver,heart and skeletal muscle.AMPK widely exists in eukaryotic cell,known as “cellularenergy regulator”.It reports that AMPK can inhibit NOX and reduce the production ofROS.AMPK has a protective effect on hepatocytes.The activation of AMPK may be amechanism of inhibiting fatty liver and hepatic fibrosis.BBR has been used as antimicrobial drug for several years in Asian countries.Moreand more studies found that BBR has exhibited antioxidant and anti-inflammatory effectsand can inhibit hepatic fibrosis caused by multiple factors.Therefor,it is of great significanceto develop the hepatoprotection effect of BBRand it’s mechanism of action.Objective:The present study used CCl4-induced liver injury model and CFSC-2G toinvestigate the role of BBR on liver injury and to investigate the correlation between hepatoprotective effect and AMPK，ROS.This study evaluated the possibility of BBR asclinical hepatoprotective drug and settled theoretical basis to the clinical application ofBerberine.Methods：Serum level of ALT,AST andALP was dected.SOD and MDAwas alsodected。Histopathological change was observed by HE stain and immunohistochemical stain.Expressing level of AMPK、NOX4、Akt、TGF-β1、α-SMA was evaluated by Western blotmethod. Inhibition effects of Berbenine on growth of CFSC-2G cell line were evaluated bySRB.Expression of ROS was measured by ELISA method.We explored the mechanism ofthe protective effect of BBR on liver injury through all above indicators.Results：1.Inhibition effect of BBR on CCl4-induced liver injury：BBRcan reduce theserum level of ALT,AST and ALP significantly.HE stain and immunohistochemical stainshow that BBR can improve CCl4-induced histopathology changes obviously.2. Effect of BBR on SOD and MDA in liver：BBR can reduce the level of MDA，andimprove SOD level.3. Inhibition effect of BBR on the proliferation of CFSC-2G cell line:The inhibitioneffect of BBR on CFSC-2G proliferation enhanced with the increase in concentration andtime.BBRwith the concentration greater than12.5μmol/L can inhibit cell proliferation after24h incubation.4. Effect on ROS level：BBR can inhibit the production of ROS in CFSC-2G.(P<0.05)5. Effect on expressing of related protein：expression of NOX4，p-Akt and TGF-β1inliver tissue of BBR group was decreased significantly compared with model group；expression of p-AMPK in liver tissue of BBR was higher than that of modelgroup;expression of α-SMA and p-Akt in BBR treated CFSC-2G decreased significantlythan control;expression of p-AMPK in BBR treated CFSC-2G was significantly higherthan that of control.Conclusion：BBR has a significant inhibitory effect on liver injury.BBR can inhibit theproliferation and activation of HSCs.The protective effect of BBR may be related to theincrease in p-AMPK, inhibition of NOX,decreased TGF-β1,cellular ROS,and p-Akt.