目的探讨柴胡皂苷D调控CaMKKβ/AMPK信号通路,在功能性消化不良中对胃肠道Cajal间质细胞(Interstitial cells of Cajal,ICCs)细胞自噬的作用及机制。方法分离大鼠原代ICCs细胞,谷氨酸刺激构建ICCs自噬模型,免疫荧光检测Ca2+水平。将原代...目的探讨柴胡皂苷D调控CaMKKβ/AMPK信号通路,在功能性消化不良中对胃肠道Cajal间质细胞(Interstitial cells of Cajal,ICCs)细胞自噬的作用及机制。方法分离大鼠原代ICCs细胞,谷氨酸刺激构建ICCs自噬模型,免疫荧光检测Ca2+水平。将原代ICCs细胞分为对照组、模型组、模型+柴胡皂苷D组、模型+CaMKKβ抑制剂组、模型+柴胡皂苷D+CaMKKβ抑制剂组。透射电镜观察自噬体超微结构,ELISA检测Ghrelin和SP的水平,免疫荧光检测Ca2+和LC-3Ⅱ的表达,Western blot检测LC-3Ⅱ/Ⅰ、CaMKKβ、p-AMPK、Drp1、MFN2、IP3R和RyR的蛋白表达水平。结果谷氨酸诱导的模型组ICCs中LC-3Ⅱ荧光表达增强。柴胡皂苷D干预可降低Ca2+浓度,降低CaMKKβ、AMPK和MFN2水平(P<0.01),增加LC-3Ⅱ/Ⅰ、IP3R、RyR、Drp1、Ghrelin和SP水平(P<0.01)。柴胡皂苷D联合CaMKKβ抑制剂STO-609干预后效果更显著。结论柴胡皂苷D可通过CaMKKβ/AMPK信号通路介导Ca2+外流,影响ICCs细胞过度自噬及胃肠动力相关因子的表达。展开更多
多项研究发现,目前近视的患病率在全球范围内成爆发性增长,且逐渐呈低龄化趋势。伴随着近视进展,屈光度不断增加、眼轴进行性的延长会引起一系列的眼部并发症,例如视网膜萎缩、视网膜脱离、视网膜变薄及撕裂等。Ca^(2+)/钙调蛋白活化的...多项研究发现,目前近视的患病率在全球范围内成爆发性增长,且逐渐呈低龄化趋势。伴随着近视进展,屈光度不断增加、眼轴进行性的延长会引起一系列的眼部并发症,例如视网膜萎缩、视网膜脱离、视网膜变薄及撕裂等。Ca^(2+)/钙调蛋白活化的蛋白激酶-β(Ca^(2+)/calmodulin-dependent protein kinase kinaseβ,CaMKKβ)/腺苷单磷酸激活的蛋白激酶(adenosine monophosphate-activated protein kinase,AMPK)信号通路是调节细胞能量代谢的经典途径之一,CaMKKβ响应于Ca^(2+)的增加而通过磷酸化激活AMPK,进一步激活自噬。近期研究发现近视视网膜病变与CaMKKβ/AMPK信号通路密切相关,故本文将主要探讨和介绍近视患者的视网膜病变与CaMKKβ/AMPK信号通路的相关性。展开更多
This study aimed to investigate the effect and mechanisms of Ephedra Herb(EH)extract on adriamycin-induced nephrotic syndrome(NS),providing an experimental basis for the clinical treatment of NS.Hematoxylin and eosin ...This study aimed to investigate the effect and mechanisms of Ephedra Herb(EH)extract on adriamycin-induced nephrotic syndrome(NS),providing an experimental basis for the clinical treatment of NS.Hematoxylin and eosin staining,creatinine,urea nitrogen,and kidn injury molecule-1 were used to evaluate the activities of EH extract on renal function.The levels of inflammatory factors and oxidative stress were detected by kits.The levels of reactive oxygen species,immune cells,and apoptosis were measured by flow cytometry.A network pharmacological approach was used to predict the potential targets and mechanisms of EH extract in the treatment of NS.The protein levels of apoptosis-related proteins and CAMKK2,p-CAMKK2,AMPK,p-AMPK,mTOR and p-mTOR in the kidneys were detected by Western blot.The effective material basis of EH extract was screened by MTT assay.The AMPK pathway inhibitor(compound C,CC)was added to investigate the effect of the potent material basis on adriamycin-induced cell injury.EH extract significantly improved renal injury and relieve inflammation,oxidative stress,and apoptosis in rats.Network pharmacology and Western blot results showed that the effect of EH extract on NS may be associated with the CAMKK2/AMPK/mTOR signaling pathway.Moreover,methylephedrine significantly ameliorated adriamycin-induced NRK-52e cell injury.Methylephedrine also significantly improved the phosphorylation of AMPK and mTOR,which were blocked by CC.In sum,EH extract may ameliorate renal injury via the CAMKK2/AMPK/mTOR signaling pathway.Moreover,methylephedrine may be one of the material bases of EH extract.展开更多
This study was designed to investigate the effect of 002C-3, a derivative of magnolol, on transient cerebral middle occlusion （tMCAO） in a mice model and to identify the underlying mechanisms. 002C-3 （100 and 150 p...This study was designed to investigate the effect of 002C-3, a derivative of magnolol, on transient cerebral middle occlusion （tMCAO） in a mice model and to identify the underlying mechanisms. 002C-3 （100 and 150 pg/kg, i.v. after ending occlusion） significantly reduced neurological deficit scores, infarct volumes, and brain water contents after 1.5 h MCAO and 24 h reperfusion. 002C-3 （75 150μg/kg） decreased the exudation of Evans blue from brain capillaries. 002C-3 （100 μg/kg） significantly inhibited the activity of MMP-9 and MMP-2 in the injured hemisphere. 002C-3 decreased the expression of autophagy-associated proteins, Beclin-1 and LC3B-Ⅱ, and increased the level of p62 in injured hemisphere. 002C-3 （100 pg/kg） significantly increased the expression of p-CaMKIV and p-HDAC4 in injured hemisphere. In conclusion, 002C-3 shows a neuroprotective effect on tMCAO injury in mice, and its mechanisms may be associated with alleviation of blood-brain barrier damage caused by the activation of MMPs, inhibition of autophagy, and stimulation of calcium signals related to cell survival. These findings suggest that 002C-3 is a neuroprotective agent that acts on multiple pathways.展开更多
文摘目的探讨柴胡皂苷D调控CaMKKβ/AMPK信号通路,在功能性消化不良中对胃肠道Cajal间质细胞(Interstitial cells of Cajal,ICCs)细胞自噬的作用及机制。方法分离大鼠原代ICCs细胞,谷氨酸刺激构建ICCs自噬模型,免疫荧光检测Ca2+水平。将原代ICCs细胞分为对照组、模型组、模型+柴胡皂苷D组、模型+CaMKKβ抑制剂组、模型+柴胡皂苷D+CaMKKβ抑制剂组。透射电镜观察自噬体超微结构,ELISA检测Ghrelin和SP的水平,免疫荧光检测Ca2+和LC-3Ⅱ的表达,Western blot检测LC-3Ⅱ/Ⅰ、CaMKKβ、p-AMPK、Drp1、MFN2、IP3R和RyR的蛋白表达水平。结果谷氨酸诱导的模型组ICCs中LC-3Ⅱ荧光表达增强。柴胡皂苷D干预可降低Ca2+浓度,降低CaMKKβ、AMPK和MFN2水平(P<0.01),增加LC-3Ⅱ/Ⅰ、IP3R、RyR、Drp1、Ghrelin和SP水平(P<0.01)。柴胡皂苷D联合CaMKKβ抑制剂STO-609干预后效果更显著。结论柴胡皂苷D可通过CaMKKβ/AMPK信号通路介导Ca2+外流,影响ICCs细胞过度自噬及胃肠动力相关因子的表达。
文摘多项研究发现,目前近视的患病率在全球范围内成爆发性增长,且逐渐呈低龄化趋势。伴随着近视进展,屈光度不断增加、眼轴进行性的延长会引起一系列的眼部并发症,例如视网膜萎缩、视网膜脱离、视网膜变薄及撕裂等。Ca^(2+)/钙调蛋白活化的蛋白激酶-β(Ca^(2+)/calmodulin-dependent protein kinase kinaseβ,CaMKKβ)/腺苷单磷酸激活的蛋白激酶(adenosine monophosphate-activated protein kinase,AMPK)信号通路是调节细胞能量代谢的经典途径之一,CaMKKβ响应于Ca^(2+)的增加而通过磷酸化激活AMPK,进一步激活自噬。近期研究发现近视视网膜病变与CaMKKβ/AMPK信号通路密切相关,故本文将主要探讨和介绍近视患者的视网膜病变与CaMKKβ/AMPK信号通路的相关性。
基金the National Key Research and Development Project(Nos.2019YFC1708802 and 2017YFC1702800)Henan province high-level personnel special support“ZhongYuan One Thousand People Pla”,Zhongyuan Leading Talent(No.ZYQR201810080)the Major Science and Technology Projects in Henan Province:Study on the key technology for quality control and the key characteristics of Rehmannia glutinosa,Dioscorea opposita Thunb.and Achyranthes bidentata Blume from Henan Province(No.171100310500).
文摘This study aimed to investigate the effect and mechanisms of Ephedra Herb(EH)extract on adriamycin-induced nephrotic syndrome(NS),providing an experimental basis for the clinical treatment of NS.Hematoxylin and eosin staining,creatinine,urea nitrogen,and kidn injury molecule-1 were used to evaluate the activities of EH extract on renal function.The levels of inflammatory factors and oxidative stress were detected by kits.The levels of reactive oxygen species,immune cells,and apoptosis were measured by flow cytometry.A network pharmacological approach was used to predict the potential targets and mechanisms of EH extract in the treatment of NS.The protein levels of apoptosis-related proteins and CAMKK2,p-CAMKK2,AMPK,p-AMPK,mTOR and p-mTOR in the kidneys were detected by Western blot.The effective material basis of EH extract was screened by MTT assay.The AMPK pathway inhibitor(compound C,CC)was added to investigate the effect of the potent material basis on adriamycin-induced cell injury.EH extract significantly improved renal injury and relieve inflammation,oxidative stress,and apoptosis in rats.Network pharmacology and Western blot results showed that the effect of EH extract on NS may be associated with the CAMKK2/AMPK/mTOR signaling pathway.Moreover,methylephedrine significantly ameliorated adriamycin-induced NRK-52e cell injury.Methylephedrine also significantly improved the phosphorylation of AMPK and mTOR,which were blocked by CC.In sum,EH extract may ameliorate renal injury via the CAMKK2/AMPK/mTOR signaling pathway.Moreover,methylephedrine may be one of the material bases of EH extract.
基金National Natural Science Foundation of China(Grant No.81302763,81573333)Beijing Natural Science Foundation(Grant No.7144218)
文摘This study was designed to investigate the effect of 002C-3, a derivative of magnolol, on transient cerebral middle occlusion （tMCAO） in a mice model and to identify the underlying mechanisms. 002C-3 （100 and 150 pg/kg, i.v. after ending occlusion） significantly reduced neurological deficit scores, infarct volumes, and brain water contents after 1.5 h MCAO and 24 h reperfusion. 002C-3 （75 150μg/kg） decreased the exudation of Evans blue from brain capillaries. 002C-3 （100 μg/kg） significantly inhibited the activity of MMP-9 and MMP-2 in the injured hemisphere. 002C-3 decreased the expression of autophagy-associated proteins, Beclin-1 and LC3B-Ⅱ, and increased the level of p62 in injured hemisphere. 002C-3 （100 pg/kg） significantly increased the expression of p-CaMKIV and p-HDAC4 in injured hemisphere. In conclusion, 002C-3 shows a neuroprotective effect on tMCAO injury in mice, and its mechanisms may be associated with alleviation of blood-brain barrier damage caused by the activation of MMPs, inhibition of autophagy, and stimulation of calcium signals related to cell survival. These findings suggest that 002C-3 is a neuroprotective agent that acts on multiple pathways.
