[Effects of Gualou Xiebai Banxia decoction on blood lipid content, oxidative stress and ox-LDL/Lox-1 pathway in ApoE-/- mice].

To observe the functions of Gualou Xiebai Banxia decoction（GXBD） on regulating lipid metabolism, anti-oxidation, and interposing ox-LDL/Lox-1 pathway, and to explore its anti-atherosclerosis (AS) mechanisms. AS models were established by using 42 Apo-E-/- male mice with high fat diet. AS model mice were randomly divided into the model group, simvastatin group, and GXBD high and low dose groups. C57BL/6J male mice were used as the normal control group, n=10 and the treatment lasted for 8 weeks. The levels of TC, TG, LDL-C, HDL-C, SOD, MDA, GSH-px, and ox-LDL in blood serum were tested 24 h after the last administration. The changes of aortic tissues structure were observed by HE staining; the expression levels of Lox-1 protein and the expression levels of mRNA were detected by Western blot and PCR respectively.Results showed that the blood lipid levels and MDA, ox-LDL levels in blood serum of model group were significantly higher than those in the normal control group, but SOD, GSH-px levels were significantly lower than those in the normal control group, and the Lox-1 protein and mRNA expression levels were also significantly higher than those in the control group(P<0.05), namely aortic atherosclerosis lesions were obvious in model group.The levels of blood lipid and MDA, ox-LDL of GXBD high and low dose groups and simvastatin group were significantly lower than those in model group, while SOD, GSH-px levels were significantly higher than those in model group, and Lox-1 protein and mRNA expression levels were significantly lower than those in model group(P<0.05), namely the aortic atherosclerosis lesions were significantly relieved. The above results indicated that GXBD was capable of modulating blood lipid, anti-oxidation, and inhibiting the expression of Lox-1, and interposing ox-LDL/Lox-1 pathway in the AS model Apo-E-/- mice, which may be one of the mechanisms of anti-atherosclerosis.

Fuzheng Huayu recipe alleviates hepatic fibrosis via inhibiting TNF-α induced hepatocyte apoptosis.

BACKGROUND: What was the relationship of Fuzheng Huayu recipe (FZHY) inhibiting hepatocyte apoptosis and HSC activation at different stage of liver fibrosis? In order to answer this question, the study was carried out to dynamically observe FZHY's effect on hepatocyte apoptosis and HSC activation and further explored underling mechanism of FZHY against hepatocyte apoptosis. METHODS: Mice were randomly divided into four groups: normal, model, FZHY, and N-acetylcystein (NAC) groups. Acute hepatic injury and liver fibrosis in mice were induced by CCl4. Three days before the first CCl4 injection, treatment with FZHY powder or NAC respectively was started. In vitro, primary hepatocytes were pretreated with FZHY medicated serum or Z-VAD-FMK and then incubated with ActD and TNF-α. Primary HSCs were treated with DNA from apoptotic hepatocytes incubated by Act D/TNF-α or FZHY medicated. Liver sections were analyzed for HE staining and immunohistochemical evaluation of apoptosis. Serum ALT and AST, Alb content and TNF-α expression in liver tissue were detected. Hyp content was assayed and collagen deposition was visualized. Expressions of α-SMA and type I collagen were analyzed by immunofluorescence and immunoblotting. Flow cytometry, immunofluorescence, and DNA ladder for hepatocyte apoptosis and immunoblotting for TNF-R1, Bcl-2 and Bax were also analyzed. RESULTS: Mice showed characteristic features of massive hepatocytes apoptosis in early stage of liver injury and developed severe hepatic fibrosis in later phase. FZHY treatment significantly alleviated acute liver injury and hepatocyte apoptosis, and inhibited liver fibrosis by decreasing α-SMA expression and hepatic Hyp content. In vitro, primary hepatocytes were induced by TNF-α and Act D. The anti-apoptotic effect of FZHY was generated by reducing TNFR1 expression and balancing the expressions of Bcl-2 and Bax. Meanwhile, the nuclear DNA from apoptotic hepatocytes stimulated HSC activation in a dose dependent manner, and the DNA from apoptotic hepatocytes treated with FZHY or Z-VAD-FMK reduced HSC activation and type I collagen expression. CONCLUSION: These findings suggested that FZHY suppressed hepatocyte apoptosis through regulating mediators in death receptor and mitochondrial pathways, and the effect of FZHY on hepatocyte apoptosis might play an important role in inhibiting liver fibrosis.

[Effects of Chinese herbal medicine Fuzheng Huayu recipe and its components against hepatocyte apoptosis in mice with hepatic injury].

OBJECTIVE: To investigate the effects and characteristics of Fuzheng Huayu recipe, a compound Chinese herbal medicine, and its decomposed therapies against hepatocyte apoptosis in mice with hepatic injury. METHODS: A total of 50 male BALB/c mice were randomly divided into 5 groups: control group, untreated group, Fuzheng Huayu recipe group, Fuzheng recipe group and Huayu recipe group. Hepatocyte apoptosis in mice was induced by intraperitoneal injection of lipopolysaccharide (10 µg/kg) and galactosamine (900 mg/kg). The mice in drug-treated groups were administered with Fuzheng Huayu recipe, Fuzheng recipe and Huayu recipe by garbage respectively 3 days before injection of lipopolysaccharide and galactosamine. The mice were sacrificed 6 hours after the administration of lipopolysaccharide and galactosamine. The activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and superoxide dismutase (SOD) and content of malondialdehyde (MDA) were examined by colorimetric method. Inflammation and necrosis in liver tissue were observed with hematoxylin-eosin staining. Hepatocyte apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining. Protein expression of tumor necrosis factor receptor type I (TNFR1) was analyzed with Western blotting. Expression of tumor necrosis factor-α (TNF-α) mRNA was analyzed with real-time fluorescent quantitative polymerase chain reaction. RESULTS: Compared with the untreated group, Fuzheng Huayu recipe, Fuzheng recipe and Huayu recipe attenuated hepatocyte apoptosis, decreased the serum ALT and AST activities and MDA content, and improved the SOD activity in liver tissues. Of the three groups, the effect of Fuzheng Huayu recipe group was the best, Fuzheng recipe group was better than Huayu recipe group. Compared with the untreated group, all drugs exerted good effects in decreasing TNF-α mRNA expression, and Fuzheng Huyu recipe was better than the others. The expression of TNFR1 protein in the untreated group was increased with the occurrence of hepatocyte apoptosis, however, only Fuzheng Huayu recipe decreased the elevated TNFR1 protein expression. CONCLUSION: Fuzheng recipe exerts a good effect in attenuating hepatocyte apoptosis in vivo, which has synergistic effects with Huayu recipe in attenuating hepatocyte apoptosis, and the potential mechanism is partially due to their different effects in alleviating oxidative stress in liver and down-regulating the expression of TNFR1 protein.