目的:从用蛋白质组学方法已经筛选出的不同运动应激条件下心肌的目标蛋白中,选择与心肌收缩和与心肌收缩调节有关的目标蛋白质。方法:利用双向凝胶电泳技术检测,比较不同运动刺激条件下心室肌和心房肌的总蛋白(与各自的对照组比较),建...目的:从用蛋白质组学方法已经筛选出的不同运动应激条件下心肌的目标蛋白中,选择与心肌收缩和与心肌收缩调节有关的目标蛋白质。方法:利用双向凝胶电泳技术检测,比较不同运动刺激条件下心室肌和心房肌的总蛋白(与各自的对照组比较),建立二维凝胶电泳(two-di mensional gel electrophoresis,2-DE)图谱。从多个差异蛋白质点中选择出3个表达明显变化的蛋白质点用MALDI-TOF-TOF质谱仪进行鉴定。结果:1702、2701、0201号点在不同负荷运动后其表达发生不同变化,1702、2701、0201号点在一次性力竭和递增负荷大运动后表达变化明显大于中等强度运动后其表达的变化,且3个点在不同负荷运动后的心室肌中的变化比在心房肌中的变化更为明显。经质谱鉴定,1702和2701号点为α-肌球蛋白重链(α-MHC),0201号点为原肌球蛋白1(tropomyosin-1,TM1)。结论:1)α-肌球蛋白重链、心肌原肌球蛋白1对运动应激的敏感性高,可作为新的研究运动应激的心脏生物标志物。2)力竭运动和递增负荷大强度运动对心肌收缩功能可能产生负面影响,而中等强度运动对心肌收缩功能没有产生不良影响,甚至会促进其收缩能力增强。展开更多
目的探讨丹参多酚酸盐对心力衰竭(心衰)大鼠心肌肌球蛋白重链(myosin heavy chain,MHC)的影响。方法采用随机数字表法将60只雄性SD大鼠分成6组:正常对照组、模型组、卡托普利组、丹参多酚酸盐低剂量组、丹参多酚酸盐高剂量组、卡托普利...目的探讨丹参多酚酸盐对心力衰竭(心衰)大鼠心肌肌球蛋白重链(myosin heavy chain,MHC)的影响。方法采用随机数字表法将60只雄性SD大鼠分成6组:正常对照组、模型组、卡托普利组、丹参多酚酸盐低剂量组、丹参多酚酸盐高剂量组、卡托普利和丹参多酚酸盐高剂量联合组(下称中西药联合组),每组10只。除正常对照组外,其余大鼠均采用阿霉素腹腔注射法制备心衰模型,正常对照组大鼠给予等体积生理盐水腹腔注射,每周1次,共6周。从注射阿霉素第5周开始,各用药组给药干预,卡托普利组予100 mg/(kg·d)的卡托普利水溶液灌胃;丹参多酚酸盐低、高剂量组分别以24.219 mg/(kg·d)及48.438 mg/(kg·d)的丹参多酚酸盐溶于2 m L 5%葡萄糖溶液中腹腔注射。中西药联合组予高剂量的丹参多酚酸盐腹腔注射,同时予卡托普利水溶液灌胃,模型组予腹腔注射生理盐水2 m L,每日1次,均连续干预8周。采用生物信号采集处理系统检测心功能、心肌肥厚指数;荧光定量PCR法检测心肌α-MHC及β-MHC mRNA表达;Western blot法检测心肌蛋白激酶C(protein kinase C,PKC)的表达。结果与正常对照组比较,模型组心脏质量指数(heart mass index,HMI)及左室质量指数(left ventricular mass index,LVMI)明显增高(P<0.01);与模型组比较,丹参多酚酸盐高剂量组、卡托普利组、中西药联合组HMI及LVMI均下降(P<0.05,P<0.01),中西药联合组较卡托普利组下降更明显(P<0.05)。与正常对照组比较,模型组心肌组织α-MHC mRNA水平降低,β-MHC mRNA水平及心肌组织PKC表达升高(P<0.01);与模型组比较,丹参多酚酸盐高剂量组、卡托普利组及中西药联合组心肌组织α-MHC mRNA水平升高,β-MHC mRNA水平及心肌组织PKC表达水平均降低(P<0.05,P<0.01),且中西药联合组与卡托普利组比较,差异有统计学意义(P<0.05)。结论丹参多酚酸盐能上调α-MHC并下调β-MHC mRNA水平,其机制可能降低PKC的表达有关。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
文摘目的:从用蛋白质组学方法已经筛选出的不同运动应激条件下心肌的目标蛋白中,选择与心肌收缩和与心肌收缩调节有关的目标蛋白质。方法:利用双向凝胶电泳技术检测,比较不同运动刺激条件下心室肌和心房肌的总蛋白(与各自的对照组比较),建立二维凝胶电泳(two-di mensional gel electrophoresis,2-DE)图谱。从多个差异蛋白质点中选择出3个表达明显变化的蛋白质点用MALDI-TOF-TOF质谱仪进行鉴定。结果:1702、2701、0201号点在不同负荷运动后其表达发生不同变化,1702、2701、0201号点在一次性力竭和递增负荷大运动后表达变化明显大于中等强度运动后其表达的变化,且3个点在不同负荷运动后的心室肌中的变化比在心房肌中的变化更为明显。经质谱鉴定,1702和2701号点为α-肌球蛋白重链(α-MHC),0201号点为原肌球蛋白1(tropomyosin-1,TM1)。结论:1)α-肌球蛋白重链、心肌原肌球蛋白1对运动应激的敏感性高,可作为新的研究运动应激的心脏生物标志物。2)力竭运动和递增负荷大强度运动对心肌收缩功能可能产生负面影响,而中等强度运动对心肌收缩功能没有产生不良影响,甚至会促进其收缩能力增强。
文摘目的探讨丹参多酚酸盐对心力衰竭(心衰)大鼠心肌肌球蛋白重链(myosin heavy chain,MHC)的影响。方法采用随机数字表法将60只雄性SD大鼠分成6组:正常对照组、模型组、卡托普利组、丹参多酚酸盐低剂量组、丹参多酚酸盐高剂量组、卡托普利和丹参多酚酸盐高剂量联合组(下称中西药联合组),每组10只。除正常对照组外,其余大鼠均采用阿霉素腹腔注射法制备心衰模型,正常对照组大鼠给予等体积生理盐水腹腔注射,每周1次,共6周。从注射阿霉素第5周开始,各用药组给药干预,卡托普利组予100 mg/(kg·d)的卡托普利水溶液灌胃;丹参多酚酸盐低、高剂量组分别以24.219 mg/(kg·d)及48.438 mg/(kg·d)的丹参多酚酸盐溶于2 m L 5%葡萄糖溶液中腹腔注射。中西药联合组予高剂量的丹参多酚酸盐腹腔注射,同时予卡托普利水溶液灌胃,模型组予腹腔注射生理盐水2 m L,每日1次,均连续干预8周。采用生物信号采集处理系统检测心功能、心肌肥厚指数;荧光定量PCR法检测心肌α-MHC及β-MHC mRNA表达;Western blot法检测心肌蛋白激酶C(protein kinase C,PKC)的表达。结果与正常对照组比较,模型组心脏质量指数(heart mass index,HMI)及左室质量指数(left ventricular mass index,LVMI)明显增高(P<0.01);与模型组比较,丹参多酚酸盐高剂量组、卡托普利组、中西药联合组HMI及LVMI均下降(P<0.05,P<0.01),中西药联合组较卡托普利组下降更明显(P<0.05)。与正常对照组比较,模型组心肌组织α-MHC mRNA水平降低,β-MHC mRNA水平及心肌组织PKC表达升高(P<0.01);与模型组比较,丹参多酚酸盐高剂量组、卡托普利组及中西药联合组心肌组织α-MHC mRNA水平升高,β-MHC mRNA水平及心肌组织PKC表达水平均降低(P<0.05,P<0.01),且中西药联合组与卡托普利组比较,差异有统计学意义(P<0.05)。结论丹参多酚酸盐能上调α-MHC并下调β-MHC mRNA水平,其机制可能降低PKC的表达有关。
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.
