The present study aimed to identify which mitogen-activated protein kinase （p38 or Jun amino-terminal kinase [JNK]） was involved in cavernosal apoptosis during the acute phase after cavernosal nerve crush injury （C...The present study aimed to identify which mitogen-activated protein kinase （p38 or Jun amino-terminal kinase [JNK]） was involved in cavernosal apoptosis during the acute phase after cavernosal nerve crush injury （CNCI） in rats to ameliorate apoptosis of cavernosal tissue, such as smooth muscle （SM）. A total of twenty 10-week-old male Sprague-Dawley rats were divided equally into two groups： sham surgery （S） and CNCI （I）. The I group approximated the clinical situation of men undergoing radical prostatectomy using two 60-second compressions of both CNs with a microsurgical vascular clamp. At 2-week postinjury, erectile response was assessed using electrostimulation. Penile tissues were harvested for immunohistochemistry analysis of alpha-SM actin （（x-SMA）, western blot analysis, and double immunofluorescence analysis of （x-SMA and phosphorylated p38 or JNK, as well as double immunofluorescent of TUNEL and phosphorylated p38 or JNK. At 2-week postinjury, the I group had a significantly lower intracavernous pressure （ICP）/mean arterial pressure （MAP） and a lower area under the curve （AUC）/MAP than the S group. The I group also exhibited decreased immunohistochemical staining of α-SMA, an increase in the number of SM cells positive for phosphorylated JNK, an increased number of apoptotic cells positive for phosphorylated JNK, and increased JNK phosphorylation compared with the S group. However, there was no significant difference in p38 phosphorylation expression or the number of SM cells positive for phosphorylated p38 between the two groups. In conclusion, our data suggest that JNK, not p38, is involved in cavernosal apoptosis during the acute phase after partial CN damage.展开更多
Background C-Jun N-terminal kinase （JNK） signaling pathway plays a critical role in cerebral ischemia. Although the mechanistic basis for this activation of JNK1/2 is uncertain, oxidative stress may play a role. The...Background C-Jun N-terminal kinase （JNK） signaling pathway plays a critical role in cerebral ischemia. Although the mechanistic basis for this activation of JNK1/2 is uncertain, oxidative stress may play a role. The purpose of this study was to investigate whether the activation of JNK1/2 is associated with the production of endogenous nitric oxide （NO）. Methods Ischemia and reperfusion （I/R） was induced by cerebral four-vessel occlusion. Sprague-Dawley （SD） rats were divided into 6 groups： sham group, I/R group, neuronal nitric oxide synthase （nNOS） inhibitor （7-nitroindazole, 7-NI） given group, inducible nitric oxide synthase （iNOS） inhibitor （2-amino-5,6-dihydro-methylthiazine, AMT） given group, sodium chloride control group, and 1% dimethyl sulfoxide （DMSO） control group. The levels of protein expression and phospho-JNK1/2 were detected by Western blotting and the survival hippocampus neurons in CA1 zone were observed by cresyl violet staining. Results The study illustrated two peaks of JNK1/2 activation occurred at 30 minutes and 3 days during reperfusion. 7-NI inhibited JNK1/2 activation during the early reperfusion, whereas AMT preferably attenuated JNK1/2 activation during the later reperfusion. Administration of 7-NI and AMT can decrease I/R-induced neuronal loss in hippocampal CA1 region. Conclusion JNK1/2 activation is associated with endogenous NO in response to ischemic insult.展开更多
目的探讨淫羊藿苷(icariin,ICA)对人口腔鳞癌细胞系CAL27细胞铁死亡的影响及其可能的作用机制。方法用不同浓度ICA处理人口腔鳞癌细胞系CAL27,用CCK-8法检测细胞增殖情况以选取最佳浓度;用不同浓度铁死亡抑制剂处理CAL27细胞,用CCK-8法...目的探讨淫羊藿苷(icariin,ICA)对人口腔鳞癌细胞系CAL27细胞铁死亡的影响及其可能的作用机制。方法用不同浓度ICA处理人口腔鳞癌细胞系CAL27,用CCK-8法检测细胞增殖情况以选取最佳浓度;用不同浓度铁死亡抑制剂处理CAL27细胞,用CCK-8法检测铁死亡抑制剂对细胞的毒性作用以选取最佳浓度;将CAL27细胞随机分为对照组、ICA组和ICA联合铁死亡抑制剂组。用Transwell小室实验检测细胞迁移和侵袭能力;检测各组细胞超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)和谷胱甘肽(glutathione,GSH)水平;用实时荧光定量聚合酶链式反应(quantitative real time polymerase chain reaction,RT-PCR)检测细胞铁死亡影响因子重链亚基溶质载体家族7成员11(solute vector family 7 member 11,SLC7A11)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)和铁蛋白重链(ferritin heavy chain,FTH1)mRNA的表达情况;用蛋白印迹法(Western blotting)检测c-Jun氨基末端激酶(c-Jun amino terminal kinase,JNK)和p53蛋白(protein 53,p53)的表达情况。结果CAL27细胞增殖能力随着ICA和铁死亡抑制剂浓度的升高而降低(P<0.05);与对照组比较,ICA组CAL27细胞侵袭数量、细胞迁移数量、SOD、GSH、SLC7A11和GPX4 mRNA的表达水平降低,MDA和FTH1 mRNA、JNK和p53蛋白的表达水平升高(P<0.05);与ICA组比较,ICA联合铁死亡抑制剂组CAL27细胞侵袭数量、细胞迁移数量、SOD、GSH、SLC7A11和GPX4 mRNA的表达水平升高,MDA、FTH1 mRNA表达水平、JNK和p53蛋白的表达水平降低(P<0.05)。结论ICA对口腔鳞癌细胞CAL27增殖、侵袭和迁移能力有一定的抑制作用,可能是通过激活JNK/p53通路提高细胞内铁死亡相关因子水平,降低细胞抗氧化能力发挥作用的。展开更多
文摘The present study aimed to identify which mitogen-activated protein kinase （p38 or Jun amino-terminal kinase [JNK]） was involved in cavernosal apoptosis during the acute phase after cavernosal nerve crush injury （CNCI） in rats to ameliorate apoptosis of cavernosal tissue, such as smooth muscle （SM）. A total of twenty 10-week-old male Sprague-Dawley rats were divided equally into two groups： sham surgery （S） and CNCI （I）. The I group approximated the clinical situation of men undergoing radical prostatectomy using two 60-second compressions of both CNs with a microsurgical vascular clamp. At 2-week postinjury, erectile response was assessed using electrostimulation. Penile tissues were harvested for immunohistochemistry analysis of alpha-SM actin （（x-SMA）, western blot analysis, and double immunofluorescence analysis of （x-SMA and phosphorylated p38 or JNK, as well as double immunofluorescent of TUNEL and phosphorylated p38 or JNK. At 2-week postinjury, the I group had a significantly lower intracavernous pressure （ICP）/mean arterial pressure （MAP） and a lower area under the curve （AUC）/MAP than the S group. The I group also exhibited decreased immunohistochemical staining of α-SMA, an increase in the number of SM cells positive for phosphorylated JNK, an increased number of apoptotic cells positive for phosphorylated JNK, and increased JNK phosphorylation compared with the S group. However, there was no significant difference in p38 phosphorylation expression or the number of SM cells positive for phosphorylated p38 between the two groups. In conclusion, our data suggest that JNK, not p38, is involved in cavernosal apoptosis during the acute phase after partial CN damage.
基金This work was supported by a grant from the Project of China Postdoctoral Science Foundation （No. 20100480568）.
文摘Background C-Jun N-terminal kinase （JNK） signaling pathway plays a critical role in cerebral ischemia. Although the mechanistic basis for this activation of JNK1/2 is uncertain, oxidative stress may play a role. The purpose of this study was to investigate whether the activation of JNK1/2 is associated with the production of endogenous nitric oxide （NO）. Methods Ischemia and reperfusion （I/R） was induced by cerebral four-vessel occlusion. Sprague-Dawley （SD） rats were divided into 6 groups： sham group, I/R group, neuronal nitric oxide synthase （nNOS） inhibitor （7-nitroindazole, 7-NI） given group, inducible nitric oxide synthase （iNOS） inhibitor （2-amino-5,6-dihydro-methylthiazine, AMT） given group, sodium chloride control group, and 1% dimethyl sulfoxide （DMSO） control group. The levels of protein expression and phospho-JNK1/2 were detected by Western blotting and the survival hippocampus neurons in CA1 zone were observed by cresyl violet staining. Results The study illustrated two peaks of JNK1/2 activation occurred at 30 minutes and 3 days during reperfusion. 7-NI inhibited JNK1/2 activation during the early reperfusion, whereas AMT preferably attenuated JNK1/2 activation during the later reperfusion. Administration of 7-NI and AMT can decrease I/R-induced neuronal loss in hippocampal CA1 region. Conclusion JNK1/2 activation is associated with endogenous NO in response to ischemic insult.
文摘目的探讨淫羊藿苷(icariin,ICA)对人口腔鳞癌细胞系CAL27细胞铁死亡的影响及其可能的作用机制。方法用不同浓度ICA处理人口腔鳞癌细胞系CAL27,用CCK-8法检测细胞增殖情况以选取最佳浓度;用不同浓度铁死亡抑制剂处理CAL27细胞,用CCK-8法检测铁死亡抑制剂对细胞的毒性作用以选取最佳浓度;将CAL27细胞随机分为对照组、ICA组和ICA联合铁死亡抑制剂组。用Transwell小室实验检测细胞迁移和侵袭能力;检测各组细胞超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)和谷胱甘肽(glutathione,GSH)水平;用实时荧光定量聚合酶链式反应(quantitative real time polymerase chain reaction,RT-PCR)检测细胞铁死亡影响因子重链亚基溶质载体家族7成员11(solute vector family 7 member 11,SLC7A11)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)和铁蛋白重链(ferritin heavy chain,FTH1)mRNA的表达情况;用蛋白印迹法(Western blotting)检测c-Jun氨基末端激酶(c-Jun amino terminal kinase,JNK)和p53蛋白(protein 53,p53)的表达情况。结果CAL27细胞增殖能力随着ICA和铁死亡抑制剂浓度的升高而降低(P<0.05);与对照组比较,ICA组CAL27细胞侵袭数量、细胞迁移数量、SOD、GSH、SLC7A11和GPX4 mRNA的表达水平降低,MDA和FTH1 mRNA、JNK和p53蛋白的表达水平升高(P<0.05);与ICA组比较,ICA联合铁死亡抑制剂组CAL27细胞侵袭数量、细胞迁移数量、SOD、GSH、SLC7A11和GPX4 mRNA的表达水平升高,MDA、FTH1 mRNA表达水平、JNK和p53蛋白的表达水平降低(P<0.05)。结论ICA对口腔鳞癌细胞CAL27增殖、侵袭和迁移能力有一定的抑制作用,可能是通过激活JNK/p53通路提高细胞内铁死亡相关因子水平,降低细胞抗氧化能力发挥作用的。
