目的探究毛兰素(Erianin)在特应性皮炎(atopic dermatitis,AD)中的作用及其在高迁移率族蛋白1(high mobility group box-1,HMGB1)/晚期糖基化终末产物受体(receptor for advanced glycation end products,RAGE)-Ras同源基因家族成员A(Ra...目的探究毛兰素(Erianin)在特应性皮炎(atopic dermatitis,AD)中的作用及其在高迁移率族蛋白1(high mobility group box-1,HMGB1)/晚期糖基化终末产物受体(receptor for advanced glycation end products,RAGE)-Ras同源基因家族成员A(Ras homolog gene family member A,RhoA)/Rho关联含卷曲螺旋结合蛋白激酶1(recombinant Rho associated coiled coil containing protein kinase 1,ROCK1)信号通路中的调控机制。方法1-氯-2,4-二硝基苯(1-Chloro-2,4-dinitrobenzene,DNCB)诱导BALB/c小鼠作为AD的模型,测量小鼠的皮肤厚度、脾和淋巴结的重量。甲苯胺蓝和HE染色检测小鼠的背部皮肤和耳朵的病理改变;ELISA检测炎症因子水平;肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)刺激HaCaT细胞建立AD体外模型;采用流式细胞术检测细胞活性氧(reactive oxygen species,ROS);免疫荧光法检测线粒体活性氧(mitochondrion reactive oxygen species,mtROS);TUNEL检测细胞凋亡情况;免疫蛋白印迹法检测HMGB1、RAGE、RhoA、ROCK1蛋白表达情况。结果在体内实验中毛兰素抑制皮肤厚度的增加,减轻脾和淋巴结重量,改善炎症细胞的浸润和肥大细胞脱颗粒,降低炎症因子水平(P<0.05)。在体外实验中,毛兰素减少TNF-α诱导的HaCaT细胞ROS、mtROS的产生(P<0.01)。毛兰素治疗后HMGB1、RAGE、RhoA及ROCK1的蛋白表达量下降(P<0.01);使用RAGE特异性阻断剂(TFA)处理r-HMGB1刺激的HaCaT细胞后,HMGB1的表达没有发生变化,RAGE、RhoA及ROCK1表达减少(P<0.01);在Rho激酶抑制剂Y-27632+r-HMGB1组中,除RAGE的表达没有降低,其余结果与TFA+r-HMGB1组相近。结论毛兰素可能通过调节HMGB1/RAGE-RhoA/ROCK1信号通路缓解特应性皮炎。展开更多
目的观察白芍总苷对肠缺血再灌注大鼠肠屏障功能的影响并探寻潜在机制。方法实验设假手术组、模型组、白芍总苷组、白芍总苷+Ras同源基因家族成员A(RhoA)拮抗剂组和白芍总苷+RhoA激动剂组,每组8只大鼠。肠缺血再灌注动物模型通过阻断肠...目的观察白芍总苷对肠缺血再灌注大鼠肠屏障功能的影响并探寻潜在机制。方法实验设假手术组、模型组、白芍总苷组、白芍总苷+Ras同源基因家族成员A(RhoA)拮抗剂组和白芍总苷+RhoA激动剂组,每组8只大鼠。肠缺血再灌注动物模型通过阻断肠系膜上动脉1 h的方法制备。通过酶联免疫吸附法(ELISA)测定血清中肠屏障功能指标,测定肠含水量,通过HE染色检查肠组织形态结构变化,逆转录聚合酶链反应(RT-PCR)法检测肠组织中RhoA、Rho相关卷曲螺旋蛋白激酶1(ROCK1)mRNA水平,蛋白质印迹法(Western blotting)检测肠组织中RhoA、ROCK1、闭锁小带蛋白1抗体(ZO-1)、Occludin、Claudins-5蛋白水平。结果经白芍总苷或白芍总苷+RhoA拮抗剂治疗4周能够显著降低肠缺血再灌注模型大鼠血清中二胺氧化酶(DAO)、D-乳酸(D-LA)、内毒素(ET)水平和肠含水量(P<0.05或P<0.01);明显改善肠组织病理性形态结构变化;降低RhoA、ROCK1 m RNA和蛋白水平,提高ZO-1、Occludin、Claudins-5蛋白水平(P<0.01)。RhoA拮抗剂能够明显增强白芍总苷对肠缺血再灌注大鼠各项指标的调控作用,RhoA激动剂则能够明显减弱白芍总苷上述作用(P<0.05或P<0.01)。结论白芍总苷具有改善肠缺血再灌注大鼠肠屏障功能的作用,其机制与抑制RhoA/ROCK1信号通路有关。展开更多
Disruption of the blood-spinal cord barrier(BSCB)is a critical event in the secondary injury following spinal cord injury(SCI).Mertk has been reported to play an important role in regulating inflammation and cytoskele...Disruption of the blood-spinal cord barrier(BSCB)is a critical event in the secondary injury following spinal cord injury(SCI).Mertk has been reported to play an important role in regulating inflammation and cytoskeletal dynamics.However,the specific involvement of Mertk in BSCB remains elusive.Here,we demonstrated a distinct role of Mertk in the repair of BSCB.Mertk expression is decreased in endothelial cells following SCI.Overexpression of Mertk upregulated tight junction proteins(TJs),reducing BSCB permeability and subsequently inhibiting inflammation and apoptosis.Ultimately,this led to enhanced neural regeneration and functional recovery.Further experiments revealed that the RhoA/Rock1/P-MLC pathway plays a key role in the effects of Mertk.These findings highlight the role of Mertk in promoting SCI recovery through its ability to mitigate BSCB permeability and may provide potential targets for SCI repair.展开更多
文摘目的探究毛兰素(Erianin)在特应性皮炎(atopic dermatitis,AD)中的作用及其在高迁移率族蛋白1(high mobility group box-1,HMGB1)/晚期糖基化终末产物受体(receptor for advanced glycation end products,RAGE)-Ras同源基因家族成员A(Ras homolog gene family member A,RhoA)/Rho关联含卷曲螺旋结合蛋白激酶1(recombinant Rho associated coiled coil containing protein kinase 1,ROCK1)信号通路中的调控机制。方法1-氯-2,4-二硝基苯(1-Chloro-2,4-dinitrobenzene,DNCB)诱导BALB/c小鼠作为AD的模型,测量小鼠的皮肤厚度、脾和淋巴结的重量。甲苯胺蓝和HE染色检测小鼠的背部皮肤和耳朵的病理改变;ELISA检测炎症因子水平;肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)刺激HaCaT细胞建立AD体外模型;采用流式细胞术检测细胞活性氧(reactive oxygen species,ROS);免疫荧光法检测线粒体活性氧(mitochondrion reactive oxygen species,mtROS);TUNEL检测细胞凋亡情况;免疫蛋白印迹法检测HMGB1、RAGE、RhoA、ROCK1蛋白表达情况。结果在体内实验中毛兰素抑制皮肤厚度的增加,减轻脾和淋巴结重量,改善炎症细胞的浸润和肥大细胞脱颗粒,降低炎症因子水平(P<0.05)。在体外实验中,毛兰素减少TNF-α诱导的HaCaT细胞ROS、mtROS的产生(P<0.01)。毛兰素治疗后HMGB1、RAGE、RhoA及ROCK1的蛋白表达量下降(P<0.01);使用RAGE特异性阻断剂(TFA)处理r-HMGB1刺激的HaCaT细胞后,HMGB1的表达没有发生变化,RAGE、RhoA及ROCK1表达减少(P<0.01);在Rho激酶抑制剂Y-27632+r-HMGB1组中,除RAGE的表达没有降低,其余结果与TFA+r-HMGB1组相近。结论毛兰素可能通过调节HMGB1/RAGE-RhoA/ROCK1信号通路缓解特应性皮炎。
文摘目的观察白芍总苷对肠缺血再灌注大鼠肠屏障功能的影响并探寻潜在机制。方法实验设假手术组、模型组、白芍总苷组、白芍总苷+Ras同源基因家族成员A(RhoA)拮抗剂组和白芍总苷+RhoA激动剂组,每组8只大鼠。肠缺血再灌注动物模型通过阻断肠系膜上动脉1 h的方法制备。通过酶联免疫吸附法(ELISA)测定血清中肠屏障功能指标,测定肠含水量,通过HE染色检查肠组织形态结构变化,逆转录聚合酶链反应(RT-PCR)法检测肠组织中RhoA、Rho相关卷曲螺旋蛋白激酶1(ROCK1)mRNA水平,蛋白质印迹法(Western blotting)检测肠组织中RhoA、ROCK1、闭锁小带蛋白1抗体(ZO-1)、Occludin、Claudins-5蛋白水平。结果经白芍总苷或白芍总苷+RhoA拮抗剂治疗4周能够显著降低肠缺血再灌注模型大鼠血清中二胺氧化酶(DAO)、D-乳酸(D-LA)、内毒素(ET)水平和肠含水量(P<0.05或P<0.01);明显改善肠组织病理性形态结构变化;降低RhoA、ROCK1 m RNA和蛋白水平,提高ZO-1、Occludin、Claudins-5蛋白水平(P<0.01)。RhoA拮抗剂能够明显增强白芍总苷对肠缺血再灌注大鼠各项指标的调控作用,RhoA激动剂则能够明显减弱白芍总苷上述作用(P<0.05或P<0.01)。结论白芍总苷具有改善肠缺血再灌注大鼠肠屏障功能的作用,其机制与抑制RhoA/ROCK1信号通路有关。
基金Natural Science Foundation of Guangdong Province(2017A030313111)National Natural Science Foundation of China(81974329).
文摘Disruption of the blood-spinal cord barrier(BSCB)is a critical event in the secondary injury following spinal cord injury(SCI).Mertk has been reported to play an important role in regulating inflammation and cytoskeletal dynamics.However,the specific involvement of Mertk in BSCB remains elusive.Here,we demonstrated a distinct role of Mertk in the repair of BSCB.Mertk expression is decreased in endothelial cells following SCI.Overexpression of Mertk upregulated tight junction proteins(TJs),reducing BSCB permeability and subsequently inhibiting inflammation and apoptosis.Ultimately,this led to enhanced neural regeneration and functional recovery.Further experiments revealed that the RhoA/Rock1/P-MLC pathway plays a key role in the effects of Mertk.These findings highlight the role of Mertk in promoting SCI recovery through its ability to mitigate BSCB permeability and may provide potential targets for SCI repair.
