1.Bile acids enhance proliferation and motility of hepatic stellate cell through regulation of p38/JNK signaling
Yining ZHANG ; Ikegami TADASHI ; Hongwei DU ; Matsuzaki YASUSHI
Journal of Clinical Pediatrics 2010;(4):301-306
Objective Bile acids (BA) facilitate cholesterol hepatic fibrosis. Although hepatic stellate cell (HSC) is one of the moot important cells during liver fibrogenesis, the effect of bile acids on HSC is rarely mentioned. Therefore,bile acids facilitate liver fibrosis through regulating activated HSC should be tested. Methods The amount of BrdU incor-poration was determined to assess the proliferation of HSC treated by bile acid. Wound-healing assay was used to determine the cellular motility. Meanwhile, the phoophorylation of p38 and JNK in HSC was detected by Western blotting. Results50 μmol/L GCDCA enhanced the HSC proliferation significantly (152.0% ± 7.1%, P < 0.05); 50 μmol/L GCDCA also induced phoophorylation of p38 and JNK (450.0% ± 12.2% of control in p38 (P < 0.01 ), 210.0% ± 15.2 % of control in JNK (P<0.05)). 50 μmol/L GCDCA aided wound healing (remaining wound area is 75.4% + 5.8% of original area, P<0.05), but this effect was inhibited by JNK (SP600125) or p38 (SB294002) inhibitor, respectively. Conclusions Bile acids enhance HSC proliferation and facilitate cellular motility through inducing phosphorylation of p38 and JNK, indicating bile acid aid liver fibrosis through regulation of p38 and JNK signaling.
2.Anti-proliferative effect of dehydroepiandrosterone and its metabolites on human tumor lines
Yanfang JIANG ; Pingwei ZHAO ; Yan TAN ; Yanqiu FANG ; Matsuzaki YASUSHI
Journal of Jilin University(Medicine Edition) 2006;0(06):-
Objective To study the inhibitory effects of dehydroepiandrosterone (DHEA) and its metabolites-dehydroepiandrosterone sulfate (DHEAs) on the proliferation of HepG2 and HT-29 and their mechanism.Methods HepG2 and HT-29 were incubated by DHEA or DHEAs with different concentrations (1,10,50,100 and 200 ?mol?L-1) for 8,24,48,72 h and routine culture was used as control.The inhibitory rate was detected by using MTT chromometry and BrdU assay respectively.The activities of 3-hydroxy-3-methylglutaryl coenzyme A reductase(HMGR),glucose -6-phosphate dehydrogenase (G6PD) and lactate dehydrogenase (LDH) were examined simultaneously.Results ①MTT chromometry:DHEA with different concentrations obviously inhibited the growth of HepG2 and HT-29 cells compared with control group(P0.05).②BrdU assay:the growth of cells were significantly inhibited by DHEA with concentrations of 50,100 and 200 ?mol?L-1,especially to HepG2 cells(P0.05).Conclusion DHEA has strong anti-proliferative effects on both HepG2 and HT-29 cell lines and inhibitory effects on the activities of G6PD or HMGR,however,DHEAs has no obvious effect.
3.Efficacy of Solifenacin on Irritable Bowel Syndrome With Diarrhea: Open-label Prospective Pilot Trial.
Yasushi FUKUSHIMA ; Hidekazu SUZUKI ; Juntaro MATSUZAKI ; Arihiro KIYOSUE ; Toshifumi HIBI
Journal of Neurogastroenterology and Motility 2012;18(3):317-323
BACKGROUND/AIMS: Solifenacin, a muscarinic type 3 receptor antagonist, is used to treat overactive bladder in adults. The aim of this study is to examine the efficacy of solifenacin on the symptomatic relief of diarrhea predominant irritable bowel syndrome (IBS-D). METHODS: A total of 20 patients with IBS-D were enrolled. After a 2-week observation period, all participants received solifenacin for 6 weeks. Subsequently, the administration of solifenacin was discontinued and ramosetron, a serotonin 3 receptor antagonist, was administered for 4 weeks. Overall improvement, the IBS-symptom severity scale (IBS-SSS), and frequency of defecation were assessed. RESULTS: Six weeks after initiation of solifenacin treatment and 4 weeks after initiation of ramosetron treatment, overall improvement was observed in 19 out of 20 (95%) and 17 out of 20 (85%) participants, respectively. At 2 weeks after initiation of solifenacin, overall improvement was observed in 16 out of 20 participants (80%). Total IBS-SSS scores at 2 and 6 weeks after the administration of solifenacin, and at 4 weeks after administration of ramosetron, were significantly lower than those at week 0. Compared to before administration, the participants' quality of life and frequency of defecation were significantly lower in all participants at 2 and 6 weeks after the administration of solifenacin and at 4 weeks after administration of ramosetron. CONCLUSIONS: The efficacy of solifenacin in the treatment of IBS with diarrhea was not inferior to that of ramosetron. Further placebo-controlled parallel studies are needed.
Adult
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Benzimidazoles
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Defecation
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Diarrhea
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Humans
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Irritable Bowel Syndrome
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Prospective Studies
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Quality of Life
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Quinuclidines
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Receptors, Serotonin, 5-HT3
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Tetrahydroisoquinolines
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Urinary Bladder
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Urinary Bladder, Overactive
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Solifenacin Succinate
4.The effect of DHEA on AKT signal pathway on transplanted Morris hepatomas in rats.
Yan-fang JIANG ; Ping-wei ZHAO ; Jun-jie QIN ; Ming-hui LI ; Matsuzaki YASUSHI ; Jun-qi NIU
Chinese Journal of Hepatology 2009;17(9):679-682
OBJECTIVETo investigate the inhibitory effect of dehydroepaimdrosterone (DHEA) on the growth of transplanted Morris hepatomas (7288CTC) in vivo in rats.
METHODS21 Buffalo rats were randomly devided into 4 groups, including one blank control group (n = 5), one group for tumor-bearing control (n = 6), and 2 experimental groups with DHEA (n = 6) or DHEA-s (n = 4). DHEA or DHEA-s was fed to the rats for 4 weeks immediately after Morris hepatomas (7288CTC) was implanted in both flanks. Phenotypes of the spleen lymphocytes were examined by flow cytometry, Akt and PTEN expression in tumor cells was detected by Western blot and immunohistochemistry.
RESULTSTumor weights of DHEA treated group were less than those of the control (P less than 0.05), the inhibitory rate was 43%. The results of Western blot and immunohistochemistry showed that in DHEA tumor group,the expression of phosphorilated Akt protein was decreased, the expression of PTEN was enhanced, the percentage of CD3 positive cells and the ratio of CD4/CD8 were increased (P less than 0.05).
CONCLUSIONDHEA can inhibit tumor growth, possibly via the inhibition of the Akt signaling pathway as well as modulating the immune function.
Animals ; Antineoplastic Agents ; pharmacology ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Dehydroepiandrosterone ; pharmacology ; Dehydroepiandrosterone Sulfate ; pharmacology ; Flow Cytometry ; Immunohistochemistry ; Liver Neoplasms, Experimental ; immunology ; metabolism ; pathology ; Neoplasm Transplantation ; PTEN Phosphohydrolase ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Random Allocation ; Rats ; Rats, Inbred BUF ; Signal Transduction ; drug effects
5.Molecular mechanisms of DHEA and DHEAs on apoptosis and cell cycle arrest via Akt pathway in hepatoma cell lines.
Yan-fang JIANG ; Ping-wei ZHAO ; Yan TAN ; Li-hua LIU ; Ming-hui LI ; Yasushi MATSUZAKI ; Jun-qi NIU
Chinese Journal of Hepatology 2007;15(6):441-444
OBJECTIVETo evaluate the anti-proliferation effects of dehydroepiandrosterone (DHEAéand DHEA sulfate (DHEAs) on tumor cells.
METHODSHuman hepatoblastoma cells (HepG2) and colon adenocarcinoma cells (HT-29) were treated with DHEA and DHEAs of various concentrations. The cells were incubated for 8, 24, 48, and 72 hours, and the proliferation, apoptosis, cell cycle and the expression of phosphorylated Akt (Thr308 and Ser473) were analyzed using MTT assay, flow cytometry, and Western blotting at different time points. The influences of an inhibitor (LY294002) and an activator (hepatic growth factor; HGF) of PI3K on the effectiveness of DHEA were determined in HepG2 cells.
RESULTSBy increasing the concentrations of DHEA (1, 10, 50, 100, 200 micromol/L), the percentages of HepG2 and HT-29 survival cells treated with DHEA at 24 h were 92.7%+/-0.9%, 84.7%+/-1.2%, 62.4%+/-0.8%, 49.5%+/-0.8%, 50.7%+/-0.3% and 92.5%+/-0.4%, 89.5%+/-0.7%, 80.5%+/-1.1%, 67.5%+/-1.5%, 70.6%+/-0.6%, respectively. Proliferations of HepG2 and HT-29 cells were significantly inhibited after 24 hours of being incubated with 100 micromol/L DHEA treatment; the inhibition effect was stronger on HepG2 cells than on HT-29 cells. The effect of DHEAs on both cell lines on cell proliferation was weaker than that of the DHEA. In the cell cycle assay, DHEA treatment induced cell arrest in G0/G1 phase in both cell lines. Apoptosis of HepG2 cells was significantly triggered (18.6%+/-2.2%) by 100 micromol/L DHEA treatment for 24 hours, but not by DHEAs. In addition, 100 and 200 micromol/L DHEA treatments for 24 hours markedly inhibited phosphorylations of Akt (Thr308 and Ser473) in HepG2 cells, and these effects were enhanced by exposing them to LY294002 and stopped by exposing them to HGF. The anti-proliferative effects of DHEA on tumor cell lines were much stronger than those of DHEAs, and they were even stronger in HepG2 cells than in HT-29 cells.
CONCLUSIONOur results suggest that the induction of apoptosis through the inhibition of Akt signaling pathway is one of the anti-proliferative mechanisms of DHEA in certain tumors, but DHEA also promotes cell-cycle arrest.
Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Proliferation ; Dehydroepiandrosterone ; pharmacology ; Dehydroepiandrosterone Sulfate ; pharmacology ; Flow Cytometry ; HT29 Cells ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction
6.Evaluation of Various Anti-oketsu Kampo Formulae—Tokakujokito,Keishibukuryogan and Tokishakuyakusan—for Microcirculation by Live Imaging
Aki HIRAYAMA ; Tsutomu TOMITA ; Hiroo YOKOTA ; Yuji KASAMAKI ; Yasushi MATSUZAKI ; Kazumasa AOYAGI
Kampo Medicine 2020;71(1):8-17
Oketsu is a characteristic pathophysiology in Kampo and traditional East Asian medicine that includes multiple aspects of hemodynamic disorder. Anti-oketsu drugs or the Kampo formulation used for oketsu show significant clinical effects on various disorders; however, their underlying mechanisms still remain unclear. We aimed to clarify the characteristics of the pharmacological effects of anti-oketsu drugs on the microcirculation using a microscopic live imaging technique. Three Kampo formulations, namely tokakujokito, keishibukuryogan, and tokishakuyakusan were orally administrated to C57BL/6 mice at a dose of 300 mg/kg diluted in distilled water. Live imaging was performed on the subcutaneous vessels of the mice, including the arteries (diameter > 50 μm), arterioles (diameter 10-50 μm) and capillaries (diameter < 10 μm). Tokakujokito widely increased erythrocyte flow velocity and blood flow volume from arteries to capillaries within 60 min of administration. The effects of keishibukuryogan on the vasodilation of the arterioles were remarkable, and continued up to 120 min after administration. The pharmacological target of tokishakuyakusan was the capillaries, increasing their erythrocyte velocity and blood flow volume;its effect was more slowly expressed than those of the other formulations. Our results clearly demonstrate the sequential and special effects of anti-oketsu drugs on hemodynamics. These differences may provide pharmacological information on the clinical usage of traditional Kampo formulations.