Herbal compound 861 regulates mRNA expression of collagen synthesis- and degradation-related genes in human hepatic stellate cells.

AIM: To identify the role of herbal compound 861 (Cpd 861) in the regulation of mRNA expression of collagen synthesis- and degradation-related genes in human hepatic stellate cells (HSCs). METHODS: mRNA levels of collagen types I and III, matrix metalloproteinase 1 (MMP-1), matrix metalloproteinase 2 (MMP-2), membrane type-1 matrix metalloproteinase (MT1-MMP), tissue inhibitor of metalloproteinase 1 (TIMP-1), and transforming growth factor beta1 (TGF-beta1) in cultured-activated HSCs treated with Cpd 861 or interferon-gamma (IFN-gamma) were determined by real-time PCR. RESULTS: Both Cpd 861 and IFN-gamma reduced the mRNA levels of collagen type III, MMP-2 and TGF-beta1. Moreover, Cpd 861 significantly enhanced the MMP-1 mRNA levels while down-regulated the TIMP-1 mRNA expression, increasing the ratio of MMP-1 to TIMP-1 to (6.3 + 0.3)- fold compared to the control group. CONCLUSION: The anti-fibrosis function of Cpd 861 may be mediated by both decreased interstitial collagen sythesis by inhibiting the transcription of collagen type III and TGF-beta1 and increased degradation of these collagens by up-regulating MMP-1 and down-regulating TIMP-1 mRNA levels.

Identification of human dopamine receptors agonists from Chinese herbs.

AIM: To find human dopamine receptors, especially D1-like receptor specific agonists from Chinese herbs as potential antihypertension drug leads. METHODS: Two D1-like receptor cell lines carrying a beta-lactamase reporter gene, and a D2 receptor cell line coexpressing a promiscuous G protein G15 were constructed using HEK293 cells. A natural compound library made from fractionated samples of herbal extracts was used for high-throughput screening (HTS) against one of the cell lines, HEK/D5R/CRE-blax. The interested hits were evaluated for their activities against various dopamine receptors. RESULTS: Fourteen hits were identified from primary screening, of which 2 of the better hit samples, HD0522 and HD0059, were selected for further material and activity analysis, and to obtain 2 compounds that appeared as 2 single peaks in HPLC, HD0522H01 and HD0059H01. HD0059H01 could activate D1, D2, and D5 receptors, with EC(50 ) values of 2.28 microg/mL, 0.85 microg/mL, and 1.41 microg/mL, respectively. HD0522H01 could only activate D1R and D5R with EC(50 ) values of 2.95 microg/mL and 8.38 microg/mL. CONCLUSION: We established cellbased assays for 3 different human dopamine receptors and identified specific agonists HD0522H01 and HD0059H01 through HTS. The specific agonist to D1-like receptors, HD0522H01, may become a new natural product-based drug lead for antihypertension treatment.

[Identification of antiviral activity of Toddalia asiatica against influenza type A virus].

OBJECTIVE: To identify antiviral activity of Toddalia asiatica against influenza virus type A in vitro. METHOD: More than two hundred Chinese medicinal herb extracts were screened for antiviral activities against influenza A/PR/8/34 (H1N1) virus using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay for virus induced cytopathic effect (CPE) in a primary screening. Positive samples were picked up and were subjected to quantitative real-time polymerase chain reaction (PCR) to quantify reduction of H1N1 virus genomic RNA. RESULT: Toddalia asiatica showed potent antiviral activities against H1N1 virus, with 50% effective concentration (EC50) value of 4.7 mg x L(-1) in MTS assay and 0.9 mg x L(-1) in quantitative PCR assay respectively. The cytotoxicity test of Toddalia asiatica generated a CC50 value of 187.2 mg x L(-1) and a selective index (SI) larger than 206 in quantitative PCR. Although the best antiviral activity of Toddalia asiatica was observed with co-treatment of influenza virus infection, it remained effective even when administrated 24 h before and after the initiation of infection. CONCLUSION: The results suggested that Toddalia asiatica compound extract could be a candidate for anti-H1N1 virus agent in the treatment of influenza.

Effects of herbal compound 861 on human hepatic stellate cell proliferation and activation.

AIM: To investigate the effects of herbal compound 861 (Cpd 861) on cell proliferation in human hepatic stellate cells (LX-2) and human hepatocellular liver carcinoma cells (HepG2), and expression of alpha-smooth muscle actin (alpha-SMA) in LX-2 cells. METHODS: LX-2 and HepG2 cells were incubated with various concentrations of Cpd 861 (0.1-0.003 mg/mL) for 1, 2, 3, 5 and 7 d. Cell proliferation was analyzed by 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. Effects of Cpd861 on the expression of alpha-SMA mRNA in LX-2 cells were measured by real-time quantitative PCR method using SYBR Green I technology. RESULTS: Cpd 861, at 0.1 mg/mL, significantly inhibited LX-2 cell proliferation (15% decrease relative to control, P<0.05) after 3 d of incubation. The inhibitory effects seemed to increase with the treatment time (25% decrease after 5 d of incubation and 35% decrease after 7 d of incubation, P<0.01). However, Cpd 861 did not affect HepG2 cell proliferation at the same concentration used for LX-2 cells. The expression levels of alpha-SMA mRNA decreased significantly when LX-2 cells were exposed to Cpd 861 for 48 h (59% decrease relative to control, P<0.05) or 72 h (60% decrease relative to control, P<0.01). CONCLUSION: Cpd 861 can significantly inhibit LX-2 cell proliferation in a dose-dependent manner, and reduce the expression levels of alpha-SMA mRNA in LX-2 cells. Since hepatic cell proliferation and high level of alpha-SMA are associated with liver fibrosis, the results suggest that Cpd 861 may be useful in the treatment of this disease.