AIM： To investigate the effects of polo-like kinase-1 （PLK1） antisense phosphorothioate oligodeoxynucleotide （ASODN） on apoptosis and cell cycle of human colon cancer cell line SW480. METHODS： After SW480 colon ...AIM： To investigate the effects of polo-like kinase-1 （PLK1） antisense phosphorothioate oligodeoxynucleotide （ASODN） on apoptosis and cell cycle of human colon cancer cell line SW480. METHODS： After SW480 colon cancer cells were transfected with PLK1 ASODN, Northern and Western blot analyses were used to examine PLK1 gene expression in cancer cells. We studied apoptosis using terminal uridine deoxynucleotidyl nick end labeling. Apoptosis and cell cycle of SW480 cells were examined by fluorescence-activated cell sorter scan. RESULTS： The levels of PLK1 mRNA and protein were greatly inhibited by PLK1 ASODN in SW480 cancer cells transfected with PLK1 ASODN. Apoptosis index （AI） induced PLK1 ASODN in a time- and dose-dependent manner. Results from FLM showed that sub-2N DNA content of transfected cancer cells was significantly increased and arrested at G2/M compared with control groups. CONCLUSION： PLK1 ASODN can induce apoptosis of human colon cancer cell line SW480.展开更多
AIM： To explore the effect of Echinococcusmultilocularis on the activation of mitogen-activated protein kinase （MAPK） signaling pathways and on livercell proliferation.METHODS： Changes in the phosphorylation of MA...AIM： To explore the effect of Echinococcusmultilocularis on the activation of mitogen-activated protein kinase （MAPK） signaling pathways and on livercell proliferation.METHODS： Changes in the phosphorylation of MAPKs and proliferating cell nuclear antigen （PCNA）expression were measured in the liver of patients withalveolar echinococcosis （AE）. MAPKs, MEK1/2 [MAPK/extracellular signal-regulated protein kinase （ERK）kinase] and ribosomal S6 kinase （RSK） phosphorylationwere detected in primary cultures of rat hepatocytesin contact in vitro with （1） E. multilocu/aris vesicle fluid（EmF）, （2）E. multilocularis-conditioned medium （EmCM）.RESULTS： In the liver of AE patients, ERK 1/2 andp38 MAPK were activated and PCNA expression wasincreased, especially in the vicinity of the metacestode.Upon exposure to EmF, p38, c-Jun N-terminal kinase（JNK） and ERK1/2 were also activated in hepatocytesin vitro, as well as MEK1/2 and RSK, in the absenceof any toxic effect. Upon exposure to EmCM, only JNKwas up-regulated.CONCLUSION： Previous studies have demonstratedan influence of the host on the MAPK cascade inE. multilocularis. Our data suggest that the reverse,i.e. parasite-derived signals efficiently acting onMAPK signaling pathways in host liver ceils, is actuallyoperating.展开更多
OBJECTIVE:To study the effect of acupuncture at Fengchi(GB 20) on the activation of myosin light chain kinase(MLCK) in the middle meningeal artery of migraine modeled rats.METHODS:Forty-four clean grade healthy female...OBJECTIVE:To study the effect of acupuncture at Fengchi(GB 20) on the activation of myosin light chain kinase(MLCK) in the middle meningeal artery of migraine modeled rats.METHODS:Forty-four clean grade healthy female Sprague-Dawley(SD) rats were randomly divided into four groups:the control group,blank control group,Fengchi(GB 20) acupuncture group,and Fengchi(GB 20) prevention group.Neurogenic inflammation of these rats was induced by electrical stimulation.The γ-^(32)P infiltration method was then used to detect MLCK activation in the middle meningeal artery,and immunocytochemistry was applied to detect the structural protein expression of MLCK.RESULTS:The migraine model was successfully established in the rats.Compared with the control group,MLCK activation was significantly decreased in the blank control group(P < 0.01).CONCLUSION:The activation of MLCK in the middle meningeal artery was increased by acupuncture at Fengchi(GB 20),indicating its effectiveness in preventing and curing on acute migraine attacks.展开更多
Objective:To explore the effect of herb-partitioned moxibustion(HPM)on tight junctions(TJs)of intestinal epithelial cells in Crohn disease(CD)mediated by tumor necrosis factor-α(TNF-α)-nuclear factor kappa B(NF-κB)...Objective:To explore the effect of herb-partitioned moxibustion(HPM)on tight junctions(TJs)of intestinal epithelial cells in Crohn disease(CD)mediated by tumor necrosis factor-α(TNF-α)-nuclear factor kappa B(NF-κB)-myosin-light-chain kinase(MLCK)pathway.Methods:Forty-eight male Sprague-Dawley rats were randomly divided into a normal control(NC)group,a model control(MC)group,an HPM group and a mesalazine(MESA)group,with 12 rats in each group.Trinitrobenzene sulfonic acid(TNBS)was administered to establish CD models.When the model was confirmed a success,the HPM group rats were treated with HPM at Tianshu(ST 25)and Qihai(CV 6),while the MESA group rats were given MESA solution by lavage.When the intervention finished,the colonic epithelial tissues were separated,purified and cultured in each group to establish the intestinal epithelial barrier model in vitro,and TNF-αwas added(100 ng/mL)in the culture medium and maintained for 24 h to establish an increased epithelial permeability model.Transepithelial electrical resistance(TEER)was used to examine the permeability of the barrier;Western blot was used to observe the expressions of the proteins related to TJs of intestinal epithelial cells mediated by TNF-α-NF-κB-MLCK pathway;immunofluorescence staining was used to observe the expressions and distributions of tight junction proteins in the intestinal epithelium.Results:After TNF-αinduction,compared with the MC+TNF-αgroup,the TEER value increased significantly in the HPM+TNF-αand MESA+TNF-αgroups(both P<0.001);the expressions of nuclear factor kappa B(NF-κB)p65,MLCK,myosin light chain(MLC),tumor necrosis factor receptor-associated factor 6(TRAF6)and receptor interaction protein-1(RIP1)decreased significantly(P<0.01 or P<0.05),and the expression of zinc finger protein A20(A20)increased significantly(P<0.01);the expressions of occludin,claudin-1,zonula occludens protein 1(ZO-1)and F-actin also increased significantly(all P<0.01).Compared with the MESA+TNF-αgroup,the expressions of MLC,occludin,claudin-1,ZO-1 and F-actin increased significantly in the HPM+TNF-αgroup(P<0.01 or P<0.05).Conclusion:HPM can protect or repair the damage of intestinal epithelial barrier in CD rats,which may be achieved through modulating the abnormal TJs in intestinal epithelium mediated by TNF-α-NF-κB-MLCK pathway.展开更多
文摘AIM： To investigate the effects of polo-like kinase-1 （PLK1） antisense phosphorothioate oligodeoxynucleotide （ASODN） on apoptosis and cell cycle of human colon cancer cell line SW480. METHODS： After SW480 colon cancer cells were transfected with PLK1 ASODN, Northern and Western blot analyses were used to examine PLK1 gene expression in cancer cells. We studied apoptosis using terminal uridine deoxynucleotidyl nick end labeling. Apoptosis and cell cycle of SW480 cells were examined by fluorescence-activated cell sorter scan. RESULTS： The levels of PLK1 mRNA and protein were greatly inhibited by PLK1 ASODN in SW480 cancer cells transfected with PLK1 ASODN. Apoptosis index （AI） induced PLK1 ASODN in a time- and dose-dependent manner. Results from FLM showed that sub-2N DNA content of transfected cancer cells was significantly increased and arrested at G2/M compared with control groups. CONCLUSION： PLK1 ASODN can induce apoptosis of human colon cancer cell line SW480.
基金Supported by A PhD grant from the French Ministry of Foreign Affairs (French Embassy in Beijing) to Ren-Yong Linby a project grant from the "Foundation Transplantation" (2005-2006)+1 种基金by a grant from NSFC, No. 30860253 and 30760239by the Xinjiang Key-Lab project grants on Echinococcosis, No. XJDX0202-2005-01 and XJDX0202-2007-04
文摘AIM： To explore the effect of Echinococcusmultilocularis on the activation of mitogen-activated protein kinase （MAPK） signaling pathways and on livercell proliferation.METHODS： Changes in the phosphorylation of MAPKs and proliferating cell nuclear antigen （PCNA）expression were measured in the liver of patients withalveolar echinococcosis （AE）. MAPKs, MEK1/2 [MAPK/extracellular signal-regulated protein kinase （ERK）kinase] and ribosomal S6 kinase （RSK） phosphorylationwere detected in primary cultures of rat hepatocytesin contact in vitro with （1） E. multilocu/aris vesicle fluid（EmF）, （2）E. multilocularis-conditioned medium （EmCM）.RESULTS： In the liver of AE patients, ERK 1/2 andp38 MAPK were activated and PCNA expression wasincreased, especially in the vicinity of the metacestode.Upon exposure to EmF, p38, c-Jun N-terminal kinase（JNK） and ERK1/2 were also activated in hepatocytesin vitro, as well as MEK1/2 and RSK, in the absenceof any toxic effect. Upon exposure to EmCM, only JNKwas up-regulated.CONCLUSION： Previous studies have demonstratedan influence of the host on the MAPK cascade inE. multilocularis. Our data suggest that the reverse,i.e. parasite-derived signals efficiently acting onMAPK signaling pathways in host liver ceils, is actuallyoperating.
基金the Beijing Natural Science Found Program[Fengchi point(GB 20)Controlling Mechanisn of MLCK Signal System of the Migraine,No.7092054
文摘OBJECTIVE:To study the effect of acupuncture at Fengchi(GB 20) on the activation of myosin light chain kinase(MLCK) in the middle meningeal artery of migraine modeled rats.METHODS:Forty-four clean grade healthy female Sprague-Dawley(SD) rats were randomly divided into four groups:the control group,blank control group,Fengchi(GB 20) acupuncture group,and Fengchi(GB 20) prevention group.Neurogenic inflammation of these rats was induced by electrical stimulation.The γ-^(32)P infiltration method was then used to detect MLCK activation in the middle meningeal artery,and immunocytochemistry was applied to detect the structural protein expression of MLCK.RESULTS:The migraine model was successfully established in the rats.Compared with the control group,MLCK activation was significantly decreased in the blank control group(P < 0.01).CONCLUSION:The activation of MLCK in the middle meningeal artery was increased by acupuncture at Fengchi(GB 20),indicating its effectiveness in preventing and curing on acute migraine attacks.
基金国家自然科学基金项目,No.81674069 and No.81473757973 Program,国家重点基础研究发展计划项目,No.2015CB554500。
文摘Objective:To explore the effect of herb-partitioned moxibustion(HPM)on tight junctions(TJs)of intestinal epithelial cells in Crohn disease(CD)mediated by tumor necrosis factor-α(TNF-α)-nuclear factor kappa B(NF-κB)-myosin-light-chain kinase(MLCK)pathway.Methods:Forty-eight male Sprague-Dawley rats were randomly divided into a normal control(NC)group,a model control(MC)group,an HPM group and a mesalazine(MESA)group,with 12 rats in each group.Trinitrobenzene sulfonic acid(TNBS)was administered to establish CD models.When the model was confirmed a success,the HPM group rats were treated with HPM at Tianshu(ST 25)and Qihai(CV 6),while the MESA group rats were given MESA solution by lavage.When the intervention finished,the colonic epithelial tissues were separated,purified and cultured in each group to establish the intestinal epithelial barrier model in vitro,and TNF-αwas added(100 ng/mL)in the culture medium and maintained for 24 h to establish an increased epithelial permeability model.Transepithelial electrical resistance(TEER)was used to examine the permeability of the barrier;Western blot was used to observe the expressions of the proteins related to TJs of intestinal epithelial cells mediated by TNF-α-NF-κB-MLCK pathway;immunofluorescence staining was used to observe the expressions and distributions of tight junction proteins in the intestinal epithelium.Results:After TNF-αinduction,compared with the MC+TNF-αgroup,the TEER value increased significantly in the HPM+TNF-αand MESA+TNF-αgroups(both P<0.001);the expressions of nuclear factor kappa B(NF-κB)p65,MLCK,myosin light chain(MLC),tumor necrosis factor receptor-associated factor 6(TRAF6)and receptor interaction protein-1(RIP1)decreased significantly(P<0.01 or P<0.05),and the expression of zinc finger protein A20(A20)increased significantly(P<0.01);the expressions of occludin,claudin-1,zonula occludens protein 1(ZO-1)and F-actin also increased significantly(all P<0.01).Compared with the MESA+TNF-αgroup,the expressions of MLC,occludin,claudin-1,ZO-1 and F-actin increased significantly in the HPM+TNF-αgroup(P<0.01 or P<0.05).Conclusion:HPM can protect or repair the damage of intestinal epithelial barrier in CD rats,which may be achieved through modulating the abnormal TJs in intestinal epithelium mediated by TNF-α-NF-κB-MLCK pathway.
