AIM:To investigate the effect of zinc protoporphyrin IX on the response of hepatoma cells to cisplatin and the possible mechanism involved.METHODS:Cytotoxicity was determined using the3-(4,5-dimethylthiazol-2-yl)-2,5-...AIM:To investigate the effect of zinc protoporphyrin IX on the response of hepatoma cells to cisplatin and the possible mechanism involved.METHODS:Cytotoxicity was determined using the3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay.Apoptosis was determined by a flow cytometric assay.Western blotting was used to measure protein expression.Heme oxygenase(HO)-1 activity was measured by determining the level of bilirubin generated in isolated microsomes.Reactive oxygen species(ROS)production was monitored by flow cytometry.Caspase-3 activity was measured with a colorimetric assay kit.Mice were inoculated with 1×107 tumor cells subcutaneously into the right flanks.All mice were sacrificed 6 wk after the first treatment and tumors were weighed and measured.RESULTS:Overexpression of HO-1 in HepG2 cell line was associated with increased chemoresistance to cisdiaminedichloroplatinum(cisplatin;CDDP)compared to other cell lines in vitro.Inhibition of HO-1 expression or activity by zinc protoporphyrin IX(ZnPP IX)markedly augmented CDDP-mediated cytotoxicity towards all liver cancer cell lines in vitro and in vivo.In contrast,induction of HO-1 with hemin increased resistance of tumor cells to CDDP-mediated cytotoxicity in vitro and in vivo.Furthermore,cells treated with ZnPP IX plus CDDP exhibited marked production of intracellular ROS and caspase-3 activity,which paralleled the incidence of cell apoptosis,whereas hemin decreased cellular ROS and caspase-3 activity induced by CDDP.CONCLUSION:ZnPP IX increases cellular sensitivity and susceptibility of liver cancer cell lines to CDDP and this may represent a mechanism of increasing ROS.展开更多
AIM: To investigate the role of heme oxygenase-1 (HO-1) in pathogenesis of experimental hepatorenal syndrome (HRS). METHODS: Rats were divided into liver cirrhotic group, zinc protoporphyrin IX (ZnPP) treatment group,...AIM: To investigate the role of heme oxygenase-1 (HO-1) in pathogenesis of experimental hepatorenal syndrome (HRS). METHODS: Rats were divided into liver cirrhotic group, zinc protoporphyrin IX (ZnPP) treatment group, cobalt protoporphyrin (CoPP) treatment group and sham group. Biliary cirrhosis was established by bile duct ligation in the first three groups. Rats in the ZnPP and CoPP treatment groups received intraperitoneal injection of ZnPP and CoPP, respectively, 24 h before sample collection. Expression of HO-1 mRNA in kidney was detected by reverse-transcription polymerase chain reaction, while protein expression was determined by immunohis-tochemical analysis. Hematoxylin and eosin staining was performed to observe liver cirrhosis and renal structure. Renal artery blood flow, mean arterial pressure and portal vein pressure, 24 h total urinary volume, serum and urine sodium concentrations, and creatinine clearance rate (Ccr) were also measured.RESULTS: The HO-1 mRNA and protein expression levels in kidney, 24 h total urinary volume, renal artery blood flow, serum and urine sodium concentration and Ccr were lower in cirrhotic group than in sham group (P < 0.05). However, they were significantly lower in ZnPP treatment group than in cirrhotic group and significantly higher in CoPP treatment group than in cirrhotic group (P < 0.05). CONCLUSION: Low HO-1 expression level in kidney is an important factor for experimental HRS.展开更多
目的探讨锌原卟啉(Znpp)对高体积分数氧(高氧)肺损伤早产大鼠的影响。方法出生3 d SD早产大鼠随机分为空气对照组(Ⅰ组),高氧对照组(Ⅱ组),空气加Znpp组(Ⅲ组),高氧加Znpp组(Ⅳ组)。Ⅱ组、Ⅳ组吸入氧体积分数≥900 mL/L,Ⅲ组和Ⅳ组每日...目的探讨锌原卟啉(Znpp)对高体积分数氧(高氧)肺损伤早产大鼠的影响。方法出生3 d SD早产大鼠随机分为空气对照组(Ⅰ组),高氧对照组(Ⅱ组),空气加Znpp组(Ⅲ组),高氧加Znpp组(Ⅳ组)。Ⅱ组、Ⅳ组吸入氧体积分数≥900 mL/L,Ⅲ组和Ⅳ组每日腹腔注射Znpp 45μmol/(kg.d)。于实验第3、7天,分别检测其肺组织匀浆血红蛋白氧合酶-1(HO-1)活性、碳氧血红蛋白(HbCO)水平及支气管肺泡灌洗液TNF-α、总蛋白、丙二醛(MDA)水平,测定肺湿/干质量比值(W/D),光镜下观察其肺组织病理学变化。结果实验第3天,Ⅱ组早产大鼠HO-1活性、HbCO水平、肺W/D及TNF-α水平、总蛋白、MDA水平明显高于Ⅰ组(Pa<0.05,0.01);Ⅳ组HO-1活性、HbCO水平、肺W/D及TNF-α水平、MDA水平明显低于Ⅱ组(Pa<0.01,0.05);实验第7天,Ⅱ组HO-1活性、HbCO水平及TNF-α水平、总蛋白、MDA水平明显高于Ⅰ组(Pa<0.01);Ⅳ组HO-1活性、HbCO水平及TNF-α水平、总蛋白、MDA水平明显低于Ⅱ组(Pa<0.01,0.05)。第3天,Ⅱ组肺组织呈急性炎性反应性改变,Ⅳ组炎性反应减轻明显;第7天,Ⅱ组呈亚急性炎性反应性改变、肺间质纤维化、肺泡发育障碍等,Ⅳ组病理损害有所减轻。结论HO-1参与早产大鼠高氧肺损伤。Znpp可通过抑制HO-1活性,减轻早产鼠高氧肺损伤。展开更多
基金Supported by Xuzhou City Health Bureau Project,China,No.XZZD1128Xuzhou City Municipal Science and Technology Bureau Project,China,No.XWJ2011040
文摘AIM:To investigate the effect of zinc protoporphyrin IX on the response of hepatoma cells to cisplatin and the possible mechanism involved.METHODS:Cytotoxicity was determined using the3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay.Apoptosis was determined by a flow cytometric assay.Western blotting was used to measure protein expression.Heme oxygenase(HO)-1 activity was measured by determining the level of bilirubin generated in isolated microsomes.Reactive oxygen species(ROS)production was monitored by flow cytometry.Caspase-3 activity was measured with a colorimetric assay kit.Mice were inoculated with 1×107 tumor cells subcutaneously into the right flanks.All mice were sacrificed 6 wk after the first treatment and tumors were weighed and measured.RESULTS:Overexpression of HO-1 in HepG2 cell line was associated with increased chemoresistance to cisdiaminedichloroplatinum(cisplatin;CDDP)compared to other cell lines in vitro.Inhibition of HO-1 expression or activity by zinc protoporphyrin IX(ZnPP IX)markedly augmented CDDP-mediated cytotoxicity towards all liver cancer cell lines in vitro and in vivo.In contrast,induction of HO-1 with hemin increased resistance of tumor cells to CDDP-mediated cytotoxicity in vitro and in vivo.Furthermore,cells treated with ZnPP IX plus CDDP exhibited marked production of intracellular ROS and caspase-3 activity,which paralleled the incidence of cell apoptosis,whereas hemin decreased cellular ROS and caspase-3 activity induced by CDDP.CONCLUSION:ZnPP IX increases cellular sensitivity and susceptibility of liver cancer cell lines to CDDP and this may represent a mechanism of increasing ROS.
基金Supported by National Natural Science Foundation of China, No. 30970886Science and Technology Project of Dalian,No. 2008E13SF193
文摘AIM: To investigate the role of heme oxygenase-1 (HO-1) in pathogenesis of experimental hepatorenal syndrome (HRS). METHODS: Rats were divided into liver cirrhotic group, zinc protoporphyrin IX (ZnPP) treatment group, cobalt protoporphyrin (CoPP) treatment group and sham group. Biliary cirrhosis was established by bile duct ligation in the first three groups. Rats in the ZnPP and CoPP treatment groups received intraperitoneal injection of ZnPP and CoPP, respectively, 24 h before sample collection. Expression of HO-1 mRNA in kidney was detected by reverse-transcription polymerase chain reaction, while protein expression was determined by immunohis-tochemical analysis. Hematoxylin and eosin staining was performed to observe liver cirrhosis and renal structure. Renal artery blood flow, mean arterial pressure and portal vein pressure, 24 h total urinary volume, serum and urine sodium concentrations, and creatinine clearance rate (Ccr) were also measured.RESULTS: The HO-1 mRNA and protein expression levels in kidney, 24 h total urinary volume, renal artery blood flow, serum and urine sodium concentration and Ccr were lower in cirrhotic group than in sham group (P < 0.05). However, they were significantly lower in ZnPP treatment group than in cirrhotic group and significantly higher in CoPP treatment group than in cirrhotic group (P < 0.05). CONCLUSION: Low HO-1 expression level in kidney is an important factor for experimental HRS.
文摘目的探讨锌原卟啉(Znpp)对高体积分数氧(高氧)肺损伤早产大鼠的影响。方法出生3 d SD早产大鼠随机分为空气对照组(Ⅰ组),高氧对照组(Ⅱ组),空气加Znpp组(Ⅲ组),高氧加Znpp组(Ⅳ组)。Ⅱ组、Ⅳ组吸入氧体积分数≥900 mL/L,Ⅲ组和Ⅳ组每日腹腔注射Znpp 45μmol/(kg.d)。于实验第3、7天,分别检测其肺组织匀浆血红蛋白氧合酶-1(HO-1)活性、碳氧血红蛋白(HbCO)水平及支气管肺泡灌洗液TNF-α、总蛋白、丙二醛(MDA)水平,测定肺湿/干质量比值(W/D),光镜下观察其肺组织病理学变化。结果实验第3天,Ⅱ组早产大鼠HO-1活性、HbCO水平、肺W/D及TNF-α水平、总蛋白、MDA水平明显高于Ⅰ组(Pa<0.05,0.01);Ⅳ组HO-1活性、HbCO水平、肺W/D及TNF-α水平、MDA水平明显低于Ⅱ组(Pa<0.01,0.05);实验第7天,Ⅱ组HO-1活性、HbCO水平及TNF-α水平、总蛋白、MDA水平明显高于Ⅰ组(Pa<0.01);Ⅳ组HO-1活性、HbCO水平及TNF-α水平、总蛋白、MDA水平明显低于Ⅱ组(Pa<0.01,0.05)。第3天,Ⅱ组肺组织呈急性炎性反应性改变,Ⅳ组炎性反应减轻明显;第7天,Ⅱ组呈亚急性炎性反应性改变、肺间质纤维化、肺泡发育障碍等,Ⅳ组病理损害有所减轻。结论HO-1参与早产大鼠高氧肺损伤。Znpp可通过抑制HO-1活性,减轻早产鼠高氧肺损伤。
