The aim of this study was to evaluate pharmacological properties of ethanol extracted from Mahonia oiwakensis Hayata stems (MOS_EtOH). The pharmacological properties included antioxidant, analgesic, anti-inflammatory and hepatoprotective effects. The protoberberine alkaloid content of the MOS_EtOH was analyzed by high-performance liquid chromatography (HPLC). The results revealed that three alkaloids, berberine, palmatine and jatrorrhizine, could be identified. Moreover, the MOS_EtOH exhibited antioxidative activity using the DPPH assay (IC₅₀, 0.743 mg/mL). The DPPH radical scavenging activity of MOS_EtOH was five times higher that that of vitamin C. MOS_EtOH was also found to inhibit pain induced by acetic acid, formalin, and carrageenan inflammation. Treatment with MOS_EtOH (100 and 500 mg/kg) or silymarin (200 mg/kg) decreased the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels compared with the CCl₄-treated group. Histological evaluation showed that MOS_EtOH reduced the degree of liver injury, including vacuolization, inflammation and necrosis of hepatocytes. The anti-inflammatory and hepatoprotective effect of MOS_EtOH were found to be related to the modulation of antioxidant enzyme activity in the liver and decreases in malondialdehyde (MDA) level and nitric oxide (NO) contents. Our findings suggest that MOS_EtOH has analgesic, anti-inflammatory and hepatoprotective effects. These effects support the use of MOS_EtOH for relieving pain and inflammation in folk medicine.

Glutamate-induced excitotoxicity has been implicated in a variety of neuronal degenerative disorders. In the present study, we investigated the possible neuroprotective effects of schizandrin against apoptosis of primary cultured rat cortical cells induced by glutamate. Glutamate (10 μM) administered for 24 h decreased the expression of Bcl-2 and Bcl-X_L protein, whereas increased the expression of Bax, Bak, apoptosis inducing factor (AIF), endonuclease G (Nodo G) and endoplasmic reticulum (ER) stress of caspase-12. Pretreatment with schizandrin (100 μM) before glutamate treatment increased the Bcl-X_L and Bcl-2 expression and decreased Bax, Bak, AIF, Nodo G and caspase-12 compared with those only treated with glutamate. Furthermore, glutamate-induced phosphorylation of JNK, p38 and ERK mitogen-activated protein kinases (MAPK), and these effects were attenuated by schizandrin (100 μM) treatment. These results suggest that schizandrin possesses the neuroprotective effects. The molecular mechanisms of schizandrin against glutamate-induced apoptosis may involve the regulation of Bcl-2 family proteins expression, and ER stress through blocking the activation of JNK, ERK and p38 MAPK.

Background: Apoptin, a nonstructural protein encoded by the VP3 gene of chicken anemia virus (CAV), has been shown to not only induce apoptosis when introduced into the precursors of chicken thymocytes, but has been found to specifically kill human cancer cells, tumor cell and transformed cells without affecting the proliferation of normal cells. This tumor-specific apoptotic characteristic of the protein potentially may allow the development of a protein drug that has applications in tumor therapy. However, several major problems, which include poor expression and poor protein solubility, have hampered the production of apoptin in bacteria. Results: Significantly increased expression of recombinant full-length apoptin that originated from chicken anemia virus was demonstrated using an E. coli expression system. The CAV VP3 gene was fused with a synthetic sequence containing a trans-acting activator of transcription (TAT) protein transduction domain (PTD). The resulting construct was cloned into various different expression vectors and these were then expressed in various E. coli strains. The expression of the TAT-Apoptin in E. coli was significantly increased when TAT-Apoptin was fused with GST-tag rather than a His-tag. When the various rare amino acid codons of apoptin were optimized, the expression level of the GST-TAT-Apoptin_opt in E. coli BL21(DE3) was significantly further increased. The highest protein expression level obtained was 8.33 g/L per liter of bacterial culture after induction with 0.1 mM IPTG for 4 h at 25 °C. Moreover, approximately 90% of the expressed GST-TAT-Apoptin_opt under these conditions was soluble. After purification by GST affinity chromatography, the purified recombinant TAT-Apoptin_opt protein was used to evaluate the recombinant protein’s apoptotic activity on tumor cells. The results demonstrated that the E. coli-expressed GST-TAT-apoptin_opt showed apoptotic activity and was able to induce human premyelocytic leukemia HL-60 cells to enter apoptosis. Conclusions: On expression in E. coli, purified recombinant TAT-Apoptin_opt that has been fused to a GST tag and had its codons optimized, was found to have great potential. This protein may in the future allow the development of a therapeutic protein that is able to specifically kill tumor cells.

This study was designed to investigate the hepatoprotective potentials of the Wu-Zi-Yuan-Chung-Wan (WZYCW) using an animal model of carbon tetrachloride (CCl₄) induced liver injury. CCl₄ induced chronic liver hepatotoxicity in adult Sprague-Dawley rats. Excluding the control group, all of the rats with chronic liver fibrosis received 0.4% CCl₄ (1.5 mL/kg of body weight, ip) twice per week for 8 weeks. WZYCW (20, 100, and 500 mg/kg) and silymarin (200 mg/kg) were administered five times per week for 8 weeks. After 8 weeks, the rats were sacrificed, blood samples were obtained, and liver histological examinations were performed for subsequent assays. These results suggest that WZYCW considerably reduced Glutamic Oxaloacetic Transaminase (GOT), Glutamic Pyruvic Transaminase (GPT), Triglyceride (TG); and cholesterol activity; and the levels of malonaldehyde (MDA), nitric oxide (NO), and transforming growth factor-β₁ (TGF-β₁) in the liver. WZYCW also increased the level of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR) in liver tissue. WZYCW produced hepatoprotective and antifibrotic effects. This is the first study to demonstrate that WZYCW expressed hepatoprotective activity against CCl₄ induced acute hepatotoxicity in rat. In addition, the primary compound of WZCYW was analyzed using HPLC. The major peaks of WZCYW, including schizandrin. The results indicate that WZYCW not only enhances hepatic antioxidant enzyme activities and inhibits lipid peroxidation but also suppresses inflammatory responses in CCl₄ induced liver damage. Our findings provide evidence that WZYCW possesses a hepatoprotective activity to ameliorate chronic liver injury.

Purpose This study investigated antidepressant-like effect of Gl-E on rats exposed to UCMS and elucidated its potential mechanism of antidepressant action of Gl-E by detecting the levels of monoamines and their metabolites in the rats’ brains. Methods Sprague-Dawley rats were subjected to UCMS and treated with extract ofG. lucidum(Gl-E) and fluoxetine (FLU) for 28 consecutive days. The sucrose preference test (SPT) and open field test (OFT) were applied to assess the antidepressant effects. The levels of monoamines including serotonin (5-hydroxytryptamine, 5-HT), norepinephrine (NE) and dopamine (DA) and their metabolites in the brain of rats were measured by HPLC. Results The results revealed that 4 weeks of UCMS exposure induced significant depression-like behavior, with low 5-HT, NE and DA levels in the rats’ brains. The treatment of Gl-E (0.1 and 0.5 g/kg) or FLU significantly alleviated the behavior deficiency and body weight reduction in UCMS rats. In addition, the levels of 5-HT, NE and DA in the brain of UCMS rats were increased after the Gl-E treatment. Conclusion The present study demonstrated that Gl-E could ameliorate depression-like behaviors in UCMS rats. Monoamines played an important role in the antidepressant action of Gl-E.