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
OBJECTIVE Metastasis-associated in colon cancer-1(MACC1)is an oncogene that has been newly identified.It promotes tumor proliferation and invasion via the MET pathway.Our study investigated the effects of Saikosaponin...OBJECTIVE Metastasis-associated in colon cancer-1(MACC1)is an oncogene that has been newly identified.It promotes tumor proliferation and invasion via the MET pathway.Our study investigated the effects of Saikosaponin-b(SS-b)on the proliferation and apoptosis of HepG2 cells and its regulation on MACC1/c-Met/Akt signaling pathway.METHODS HepG2 cells were treated with SS-b(10-800 g·L^(-1))for 48 h in vitro.The CCK-8 assay was used to assess cell proliferation,and cell apoptosis was determined by Hoechst33258 staining,AnnexinⅤ/PI staining and caspase 3 assay.RT-PCR was used to examine the expression of MACC1,c-MET and hepatocyte growth factor(HGF)mR NA.MACC1 protein was detected by Western blot and immunohistochemistry.The protein expressions of p-cMET,c-MET,p-AKT,AKT,p-BAD,BAD were measured by Western blot.RESULTS SS-b inhibited the growth of HepG2 cells in dose-dependent way and induced cell apoptosis significantly.HepG2 cells showed karyopyknosis,fragmentation and fluorescence highlight in SS-b treatment group.FCM results showed that apoptosis rate of HepG2 cells increased with SS-b concentration.The immunofluorescence results showed that the MACC1 expression decreased significantly in HepG2 cells treated with SS-b.The expression levels of MACC1,c-MET and HGF mR NA in HepG2 cells were significantly inhibited by SS-b.SS-b also significantly decreased the protein expressions of MACC1,p-c-MET and p-AKT while increased the expression of p-BAD and caspase 3 in HepG2 cells(P<0.05).CONCLUSION SS-b inhibited the proliferation and induced the apoptosis of HepG2 cells by targeting the MACC1/c-Met/Akt signaling pathway.展开更多
Using the negative eigenvalue theory and the infinite order perturbation theory, a new method was developed to solve the eigenvectors of disordered systems. The result shows that eigenvectors change from the extended ...Using the negative eigenvalue theory and the infinite order perturbation theory, a new method was developed to solve the eigenvectors of disordered systems. The result shows that eigenvectors change from the extended state to the localized state with the increase of the site points and the disordered degree of the system. When electric field is exerted, the electrons transfer from one localized state to another one. The conductivity is induced by the electron transfer. The authors derive the formula of electron conductivity and find the electron hops between localized states whose energies are close to each other, whereas localized positions differ from each other greatly. At low temperature the disordered system has the character of the negative differential dependence of resistivity and temperature.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。
基金supported by Scientific and Technology Projects of Henan Province(142102310137)Science and Technology Development Project of Luoyang City(1603001A-3)
文摘OBJECTIVE Metastasis-associated in colon cancer-1(MACC1)is an oncogene that has been newly identified.It promotes tumor proliferation and invasion via the MET pathway.Our study investigated the effects of Saikosaponin-b(SS-b)on the proliferation and apoptosis of HepG2 cells and its regulation on MACC1/c-Met/Akt signaling pathway.METHODS HepG2 cells were treated with SS-b(10-800 g·L^(-1))for 48 h in vitro.The CCK-8 assay was used to assess cell proliferation,and cell apoptosis was determined by Hoechst33258 staining,AnnexinⅤ/PI staining and caspase 3 assay.RT-PCR was used to examine the expression of MACC1,c-MET and hepatocyte growth factor(HGF)mR NA.MACC1 protein was detected by Western blot and immunohistochemistry.The protein expressions of p-cMET,c-MET,p-AKT,AKT,p-BAD,BAD were measured by Western blot.RESULTS SS-b inhibited the growth of HepG2 cells in dose-dependent way and induced cell apoptosis significantly.HepG2 cells showed karyopyknosis,fragmentation and fluorescence highlight in SS-b treatment group.FCM results showed that apoptosis rate of HepG2 cells increased with SS-b concentration.The immunofluorescence results showed that the MACC1 expression decreased significantly in HepG2 cells treated with SS-b.The expression levels of MACC1,c-MET and HGF mR NA in HepG2 cells were significantly inhibited by SS-b.SS-b also significantly decreased the protein expressions of MACC1,p-c-MET and p-AKT while increased the expression of p-BAD and caspase 3 in HepG2 cells(P<0.05).CONCLUSION SS-b inhibited the proliferation and induced the apoptosis of HepG2 cells by targeting the MACC1/c-Met/Akt signaling pathway.
文摘Using the negative eigenvalue theory and the infinite order perturbation theory, a new method was developed to solve the eigenvectors of disordered systems. The result shows that eigenvectors change from the extended state to the localized state with the increase of the site points and the disordered degree of the system. When electric field is exerted, the electrons transfer from one localized state to another one. The conductivity is induced by the electron transfer. The authors derive the formula of electron conductivity and find the electron hops between localized states whose energies are close to each other, whereas localized positions differ from each other greatly. At low temperature the disordered system has the character of the negative differential dependence of resistivity and temperature.
