[Objective] This study was to investigate the role of IP3 sensitive calcium channel in the JA-induced calcium mobilization pathway.[Method] Arabidopsis thaliana leaves were labeled by Fluorescent probe Fluo-3/AM under...[Objective] This study was to investigate the role of IP3 sensitive calcium channel in the JA-induced calcium mobilization pathway.[Method] Arabidopsis thaliana leaves were labeled by Fluorescent probe Fluo-3/AM under low temperature at 4 ℃ to measure the fluorescent intensity of intracellular Ca2＋ which was pretreated with heparin on jasmonic acid（JA）-induced.[Results] When A.thaliana leaf cells were pretreated with 10,50 or 100 ng/ml heparin,intercellular free Ca2＋ fluorescence intensity was reduced in comparison with negative control.Once the heparin-pretreated A.thaliana leaf cells were stimulated with 100 μmol/L JA,intercellular Ca2＋ fluorescence intensity increased gradually and tended to be stable at a degree equivalent with that in negative control.[Conclusion] The experiment showed that the pretreatment with heparin could inhibit the increase of the intracellular Ca2＋ concentration significantly which JA-induced in leaves of Arabidopsis thaliana.展开更多
OBJECTIVE: To evaluate the anti-hepatoma effect of Calmodulin antagonist 0 - 4-ethoxyl-butyl-Berbamine (EBB), one of the berbamine derivatives. METHODS: Monotetrazolium (MTT) method was used to analyze the effect of E...OBJECTIVE: To evaluate the anti-hepatoma effect of Calmodulin antagonist 0 - 4-ethoxyl-butyl-Berbamine (EBB), one of the berbamine derivatives. METHODS: Monotetrazolium (MTT) method was used to analyze the effect of EBB on the proliferation and growth inhibition effect. Of a hepatoma cell line in vitro. A mouse hepatoma model was induced by injection of hepatoma cells (H22) in the abdominal cavity. The effect of EBB on survival at different concentrations as well as in combination with 5-FU were investigated in vivo. Flow cytometry analysis, dot blot hybridization, western blot, immunochemistry, enzyme-linked lectin assay (ELISA), trifluoperazine (TFP) and electron microscopic observation were used to study the effect of EBB on cell cycle process, P53 mRNA and protein levels, calmodulin content and ultrastractural changes of hepatoma cells. RESULTS: EBB exerts a very strong inhibitory effect on human hepatoma cell line 7402 and mouse hepatoma cell line H22 in vitro. The IC(50) value of EBB for the two cell lines are 3.312 microg/ml and 1.167 microg/ml, respectively. The sensitivity of H22 cells to 5-FU can be markedly enhanced: The IC(50) dosage of 5-Fu can be decreased from 0.75 microg/ml down to 0.15 microg/ml, when jointly administered with nontoxic dosages of EBB (IC(10)). In vivo, EBB can prolong the lifespan of mice with ascites H22 to more than three months. 64% of mice survived, while all animals in the control group died by the 18th day. When EBB (5 mg x kg(-1) x d(-1)) is jointly used with 5-FU (25 mg x ml(-1) x d(-1)), 73% of mice with ascites H22 survived, much higher than 27% in the 5-FU treated group. EBB can enhance the anti-hepatoma ability of 5-Fu treatment. EBB mechanism against hepatoma: P53 expression in the EBB treated group is substantially higher than that in the control group. EBB increased the translation of P53. As a calmodulin antagonist, EBB decreases amount of the CaM in hepatoma cells and blocked the hepatoma cell proliferation cycle at the G(2)M phase. Before the G(0)/G(1) phase, a diploid peak and apoptic cells in the treated groups were observed. CONCLUSIONS: The CaM antagonist, EBB, has a strong anti-hepatoma effect and enhances the effect of 5-FU, induces hepatoma cell to apoptosis, promotes the P53 protein expression and decreases the amount of CaM in the cytoplasm. All these results demonstrate that EBB is a new and potentially useful drug against hepatoma and should be researched further.展开更多
基金Supported by National Natural Science Foundation of China(30700428,30911130166)Natural Science Foundation of Beijing Municipality(5072009)The New Star Plan of Science and Technology in Beijing Municipality(2006B26)~~
文摘[Objective] This study was to investigate the role of IP3 sensitive calcium channel in the JA-induced calcium mobilization pathway.[Method] Arabidopsis thaliana leaves were labeled by Fluorescent probe Fluo-3/AM under low temperature at 4 ℃ to measure the fluorescent intensity of intracellular Ca2＋ which was pretreated with heparin on jasmonic acid（JA）-induced.[Results] When A.thaliana leaf cells were pretreated with 10,50 or 100 ng/ml heparin,intercellular free Ca2＋ fluorescence intensity was reduced in comparison with negative control.Once the heparin-pretreated A.thaliana leaf cells were stimulated with 100 μmol/L JA,intercellular Ca2＋ fluorescence intensity increased gradually and tended to be stable at a degree equivalent with that in negative control.[Conclusion] The experiment showed that the pretreatment with heparin could inhibit the increase of the intracellular Ca2＋ concentration significantly which JA-induced in leaves of Arabidopsis thaliana.
文摘OBJECTIVE: To evaluate the anti-hepatoma effect of Calmodulin antagonist 0 - 4-ethoxyl-butyl-Berbamine (EBB), one of the berbamine derivatives. METHODS: Monotetrazolium (MTT) method was used to analyze the effect of EBB on the proliferation and growth inhibition effect. Of a hepatoma cell line in vitro. A mouse hepatoma model was induced by injection of hepatoma cells (H22) in the abdominal cavity. The effect of EBB on survival at different concentrations as well as in combination with 5-FU were investigated in vivo. Flow cytometry analysis, dot blot hybridization, western blot, immunochemistry, enzyme-linked lectin assay (ELISA), trifluoperazine (TFP) and electron microscopic observation were used to study the effect of EBB on cell cycle process, P53 mRNA and protein levels, calmodulin content and ultrastractural changes of hepatoma cells. RESULTS: EBB exerts a very strong inhibitory effect on human hepatoma cell line 7402 and mouse hepatoma cell line H22 in vitro. The IC(50) value of EBB for the two cell lines are 3.312 microg/ml and 1.167 microg/ml, respectively. The sensitivity of H22 cells to 5-FU can be markedly enhanced: The IC(50) dosage of 5-Fu can be decreased from 0.75 microg/ml down to 0.15 microg/ml, when jointly administered with nontoxic dosages of EBB (IC(10)). In vivo, EBB can prolong the lifespan of mice with ascites H22 to more than three months. 64% of mice survived, while all animals in the control group died by the 18th day. When EBB (5 mg x kg(-1) x d(-1)) is jointly used with 5-FU (25 mg x ml(-1) x d(-1)), 73% of mice with ascites H22 survived, much higher than 27% in the 5-FU treated group. EBB can enhance the anti-hepatoma ability of 5-Fu treatment. EBB mechanism against hepatoma: P53 expression in the EBB treated group is substantially higher than that in the control group. EBB increased the translation of P53. As a calmodulin antagonist, EBB decreases amount of the CaM in hepatoma cells and blocked the hepatoma cell proliferation cycle at the G(2)M phase. Before the G(0)/G(1) phase, a diploid peak and apoptic cells in the treated groups were observed. CONCLUSIONS: The CaM antagonist, EBB, has a strong anti-hepatoma effect and enhances the effect of 5-FU, induces hepatoma cell to apoptosis, promotes the P53 protein expression and decreases the amount of CaM in the cytoplasm. All these results demonstrate that EBB is a new and potentially useful drug against hepatoma and should be researched further.
