ABSTRACT
Aim To compare the effects of two different methods of establishing non-alcoholic fatty liver disease ( NAFLD) model, and to explore a more efficient method for establishing NAFLD model that conforms to the characteristics of human disease.Methods C57BL/6J mice were randomly divided into NI) group, WD group, WD combined with CC14 group.'Hie NI) group was fed a normal maintenance diet, and WD group was fed WD.On the basis of WD feed, mice in WD + CC14 group were intraperitoneally injected with CC14 oil solution.'Hie mice were sacrificed on 6, 11 and 16 weeks after modeling.HE staining and oil red 0 staining were performed to observe the pathological changes of liver.The serum levels of ALT, AST, TG and T- CHO were detected by automatic biochemical analyzer, and the levels of IL-lp, IL-6 and TNF-a in liver homogenate were detected by ELISA.The protein expression of FAS and a-SMA was detected by Western blot.Results As the development of model, pathological results showed that NAFLD model was success-fully established by these two methods.At the same time point of modeling, compared with WD group, the liver pathology of WD + CC14 group was more serious, liver steatosis appeared since 6th week.The serum ALT, AST levels and the contents of TG and T-CHO significantly increased.Meanwhile, the levels of inflammatory cytokines obviously increased in the liver, the expression of fibrosis-associated protein a-SMA increased, and the model could progress to the stage of NASH on 16th week.The course of NAFLD in the WD group progressed slowly, and steatosis appeared on 1 1 th week, and it was further aggravated till 16th week.The pro-tein level of FAS was significantly higher than that in WD + CC14 group, and no obvious inflammatory cell infiltration was observed.Conclusions WD feed combined with CC14 to establish NAFLD model takes shorter time and exerts better effect than feeding WD a- lone.It can progress to NASH on 16th week, which can be used as an ideal method to establish NAFLD model.
ABSTRACT
<p><b>AIM</b>To investigate the effect of baicalin on the hippocampal neuronal apoptosis and the expression of HSP70 in rats with focal brain ischemia-reperfusion injury.</p><p><b>METHODS</b>One hundred and twenty male Wistar rats were randomly divided into six groups:sham operated group, ischemia-reperfusion group, nimodipine group and three baicalin groups,to which baicalin was administered at doses of 50, 100 and 200 mg x kg(-1), separately. The models of focal brain ischemia-reperfusion injury induced by middle cerebral artery occlusion (MCAO) were used in this study. HE stain was used to observe the pathological changes. Flow cytometry (FCM) was used for determination of neuronal apoptosis. HSP70 protein expression of the neurons was detected with immunohistochemistry. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of the mRNA level of HSP70.</p><p><b>RESULTS</b>Baicalin can significantly relieve the pathological changes and inhibit apoptosis in hippocampus CA1 area, and at the same time increase the expression of HSP70 and HSP70 mRNA.</p><p><b>CONCLUSION</b>Baicalin can relieve brain damage induced by focal brain ischemia-reperfusion in rats, which may be related to inhibiting the process of the neuronal apoptosis. The mechanism of antiapoptosis effect of baicalin may be related to the promotion of transcription of HSP70 mRNA and increasing the expression of the protein.</p>
Subject(s)
Animals , Male , Rats , Apoptosis , Flavonoids , Pharmacology , Flow Cytometry , HSP70 Heat-Shock Proteins , Genetics , Hippocampus , Metabolism , Pathology , Infarction, Middle Cerebral Artery , Neurons , Metabolism , Pathology , Neuroprotective Agents , Pharmacology , Plants, Medicinal , Chemistry , Pyramidal Cells , Metabolism , Pathology , RNA, Messenger , Genetics , Random Allocation , Rats, Wistar , Reperfusion Injury , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Scutellaria , ChemistryABSTRACT
<p><b>AIM</b>To observe the damages induced by hydrogen peroxide in cultured bovine cerebral microvascular endothelial cells (BCMEC) and evaluate the protective effects of hydroxyethylpuerarin on hydrogen peroxide-injured BCMEC.</p><p><b>METHODS</b>BCMEC were cultured and transferred into modified Eagle medium (MEM). The viability of cells was detected by MTT assay. Cell injury was determined by lactate dehydrogenase (LDH) activity in the extracellular medium. Flow cytometry was employed to observe the occurrence of apoptosis. Morphologic changes of cells were visualized under phase contrast and electron microscopes.</p><p><b>RESULTS</b>Hydrogen peroxide (200 micromol x L(-1) for 4 hours) inhibited the viability of cultured BCMEC and stimulated LDH release. Hydrogen peroxide (100 micromol x L(-1) for 4 hours) induced the occurrence of apoptosis. Hydroxyethylpuerarin was shown to increase the survival rate and decrease the activity of LDH of BCMEC damaged by hydrogen peroxide. Hydroxyethylpuerarin was also found to protect BCMEC against apoptosis induced by hydrogen peroxide.</p><p><b>CONCLUSION</b>Hydrogen peroxide induces BCMEC injury either by apoptosis or through necrosis. Hydroxyethylpuerarin protects BCMEC against hydrogen peroxide-induced injury in a concentration-dependent manner. Its antioxidant effects might be involved as the mechanism protection.</p>