Sorafenib inhibits liver cancer growth by decreasing mTOR, AKT, and PI3K expression.

PURPOSE: The purpose of this study was to determine the impact of sorafenib on PI3K/AKT/mTOR signaling pathway and to further define its mechanism for treating hepatocellular carcinoma (HCC). METHODS: Human SMMC-7721 hepatic carcinoma cells were treated with or without 4 µmoL/L sorafenib. SMMC- 7721 cells were harvested at various time points (0-48 hrs) and assessed for changes in PI3K, mTOR, and AKT protein and mRNA levels. RESULTS: Human SMMC-7721 hepatic tumor cells exposed to sorafenib had decreased expression of PI3K/mTOR/AKT. CONCLUSION: Sorafenib appears to inhibit hepatic tumor growth by downregulating PI3k/Akt/mTOR signaling pathway.

[Identification of the active material of anti-hepatic fibrosis from Amydae Carapax].

OBJECTIVE: To identify the active material of anti-hepatic fibrosis from Amydae Carapax. METHODS: Membrane separation technology was adopted to screen active fraction in Amydae Carapax, and the active components were isolated from the active fraction using gel chromatography and high performance liquid chromatography. The purified active components in Amydae Carapax were further analyzed using 4700 series time-of-flight mass spectrometer. RESULTS: Proteins and peptides of Amydae Carapax with molecular weight less than 6000 were proved to have biological activity. 8 components (Bj1-Bj8) were isolated from the active fraction. Bj4, Bj6 and Bj7 were screened as active components. Bj7 was further purified, resulting in 7 components (Bj701-Bj707). Bj704 and Bj707 showed significant biological activity. Mass spectrometry showed three molecular ion peaks with highest abundance, i.e. m/e 526, 542 and 572, i.e. m/e 526, 542 and 572, in Bj707 -A The amino acid sequences of above three peptide compounds were NDDY (Asn-Asp-Asp-Tyr), NPNPT (Asn-Pro-Asn-Pro-Thr), and HGRFG (His-Gly-Arg-Phe-Gly), respectively. And M572 was the most abandunt components. CONCLUSION: Three active peptide compounds of anti-hepatic fibrosis of Amydae Carapax were identified.

[Study on separation and purification of anthraquinones in radix et rhizoma rhei by D301 macroporous resin].

OBJECTIVE: The parameters of absorption and purification of total anthraquinones in Radix Et Rhizoma Rhei by D301 macroporous resin were investigated in this paper. METHOD: HPLC was used to analyze the content of total anthraquinones in Radix Et Rhizoma with emodin as control. RESULT: The appropriate adsorption conditions were: concentration of extract 0.5 g x mL(-1); pH 9; flow rate 1 BV x h(-1). When the 75% ethanol was used as elution and the concentration of HCl was 0.1 mol x L(-1), and the flow rate was 1.0 BV x h(-1), the effect of desorption was satisfactory. CONCLUSION: D301 resin provided a good effect on the exchang and absorption of total anthraquinones in Radix Et Rhizoma.

Kangxian ruangan keli inhibits hepatic stellate cell proliferation mediated by PDGF.

AIM: To investigate the effect of Kangxian ruangan keli (KXR) on hepatic stellate cell (HSC) proliferation mediated by platelet-derived growth factor (PDGF) and the underlying mechanism. METHODS: In a serum-free culture system, HSCs were treated with a KXR preparation for 24 hours, followed by stimulation with PDGF-BB for 24 hours. Then the cells were incubated again in the medium containing KXR for 3 hours stimulated with PDGF-BB for 5 minutes, and collected. The proliferation of HSC was examined using an MTT assay and flow cytometry. Tyrosine phosphorylation was detected with Western blotting and visualized by the enhanced chemiluminescent (ECL) method. RESULTS: The OD values for the HSCs growing in the media without and with addition of PDGF were 0.17+/-0.06 and 0.82+/-0.05, respectively. The PDGF-induced increase was hindered remarkably by KXR preparation in a dose-dependent manner. The reaction values for the systems with 5 mg/mL, 2.5 mg/mL and 1.25 mg/mL of KXR were 0.28+/-0.03, 0.37+/-0.02 and 0.43+/-0.04, respectively. Moreover, the percentages of S-phase cells in these KXR-containing culture systems were 10.95+/-1.35, 32.76+/-1.07 and 43.19+/-1.09, respectively, all of which were significantly lower than that in the culture free of KXR (68.24+/-2.72). In addition, the values for tyrosine-phosphorylated protein in HSCs treated with 5 mg/mL and 1.25 mg/mL of KXR were 0.1349+/-0.0072 and 0.1658+/-0.0025, respectively, which were smaller than that in the cells treated only with PDGF-BB (0.1813+/-0.0117). CONCLUSION: Within the dose range used in the present study, KXR preparation shows an inhibitory effect on HSC proliferation induced by PDGF. The mechanism of this process may involve interference with tyrosine phosphorylation mediated by PDGF.

[Hepatocyte apoptosis and the expression of Bcl-2 and Bax in Yin-jaundice rats].

OBJECTIVE: To investigate the effects of Yinchen Shufu Decoction on hepatocyte apoptosis and the expression of its apoptosis-regulating gene Bcl-2 and Bax in Yin-jaundice rats. METHODS: Forty-eight rats were divided randomly into 4 groups: the normal control group, Yin-jaundice model group, Yang-jaundice model group, Yinchen Shufu Decoction treatment group. The TUNEL assay and the immunohistochemistry assay were used to detect the apoptosis of hepatocytes and the expression of Bcl-2 and Bax in hepatocytes respectively. RESULTS: The rate of apoptosis cells in Yin-jaundice model group was higher significantly than that in Yang-jaundice model group and normal control group (P<0.01), the expression of Bcl-2 in Yinchen Shufu Decoction treatment group was higher significantly than that in Yin-jaundice model group (P<0.01 or P<0.05), and the expression of Bax in it was lower significantly than that in Ying-jaundice model group (P<0.01 or P<0.05). CONCLUSION: Yinchen Shufu Decoction can prevent hepatocyte apoptosis perhaps by up-regulating the expression of Bcl-2 and down-regulating the expression of Bax. It is one of the mechanisms of its treatment on Yin-jaundice.