Objective To investigate the effects of electroacupuncture(EA)on the expression and phosphorylation of insulin signal transduction-related proteins in the hypothalamus of insulin resistance(IR)rats.Methods There were ...Objective To investigate the effects of electroacupuncture(EA)on the expression and phosphorylation of insulin signal transduction-related proteins in the hypothalamus of insulin resistance(IR)rats.Methods There were totally seventy-five Wistar rats.Ten rats were randomly assigned to the Normal group(N).The remaining 65 rats were fed a high-fat diet,fifty successfully modeled rats were randomly divided into the Model(M),EA,Sham EA(S+EA),l-leucine(L),and l-leucine+EA(L+EA)groups,with 10 rats in each group.EA was applied at acupoints“Guanyuan(CV 4)”,“Zhongwan(CV 12)”,“Zusanli(ST 36)”,and“Fenglong(ST 40)”,with each session lasting 10 min,three times per week for 8 weeks.The S+EA group received needle insertion to a depth of≤2 mm without electrical stimulation,with the same treatment duration and same acupoint selection.Body weight,fasting blood glucose(FBG),and insulin sensitivity(glucose infusion rate,GIR)were measured.Western blot analysis was used to assess insulin receptor substrate-1(IRS-1),(Protein kinase B)Akt,glycogen synthase kinase-3β(GSK-3β),mechanistic target of rapamycin complex 1(mTORC1),and Ribosomal S6 kinase 1(S6K1),along with their phosphorylated forms.PCR was used to evaluate mRNA expression of IRS-1,Akt,and GSK-3β.Immunofluorescence was used to detect hypothalamic Akt localization.Results(1)Compared to the N group,the M group exhibited increased body weight,FBG,and phosphorylation of GSK-3β,mTORC1,and S6K1,with decreased GIR,IRS-1,Akt phosphorylation,and mRNA expression(P<0.05,P<0.01).(2)Compared to the M group,the EA and S+EA groups showed reduced body weight,FBG,GSK-3β,mTORC1,and S6K1 phosphorylation,with increased GIR,IRS-1,Akt phosphorylation,and mRNA expression(P<0.05,P<0.01).(3)Compared to the EA group,the S+EA group had higher body weight,GSK-3βphosphorylation,and mRNA expression,with reduced p-IRS-1 and p-Akt expression(P<0.05);the L and L+EA groups showed increased GSK-3β,mTORC1,and S6K1 phosphorylation,with decreased GIR,IRS-1,and Akt mRNA expression(P<0.05).(4)Compared to the L+EA group,the L group exhibited higher GSK-3β,mTORC1,and S6K1 phosphorylation,with lower GIR,Akt mRNA,and p-Akt expression(P<0.05,P<0.01).Conclusion EA positively influences body weight,glucose-lipid metabolism,and insulin sensitivity in IR rats,with regulatory effects on central insulin signal transduction-related proteins,potentially linked to its suppression of hypothalamic mTORC1/S6K1 pathway activity.展开更多
BACKGROUND As a member of the chaperonin-containing tailless complex polypeptide 1(TCP1)complex,which plays a pivotal role in ensuring the accurate folding of numerous proteins,chaperonin-containing TCP1 subunit 6A(CC...BACKGROUND As a member of the chaperonin-containing tailless complex polypeptide 1(TCP1)complex,which plays a pivotal role in ensuring the accurate folding of numerous proteins,chaperonin-containing TCP1 subunit 6A(CCT6A)participates in various physiological and pathological processes.However,its effects on cell death and cancer therapy and the underlying mechanisms need further exploration in colorectal cancer(CRC)cells.AIM To explore the effects of CCT6A on cell death and cancer therapy and the underlying mechanisms in CRC.METHODS Cell proliferation was evaluated using the MTS assay,EdU staining,and colony growth assays.The expression of CCT6A was monitored by immunoblotting and quantitative PCR.CCT6A was knocked out by CRISPR-Cas9,and overexpressed by transfecting plasmids.Autophagy was examined by immunoblotting and the mCherry-GFP-LC3 assay.To monitor apoptosis and necroptosis,immunoblotting,co-immunoprecipitation,and flow cytometry were employed.RESULTS Cisplatin(DDP)exerted cytotoxic effects on CRC cells while simultaneously downregulating the expression of CCT6A.Depletion of CCT6A amplified the cytotoxic effects of DDP,whereas overexpression of CCT6A attenuated these adverse effects.CCT6A suppressed autophagy,apoptosis,and necroptosis under both basal and DDP-treated conditions.Autophagy inhibitors significantly enhanced the cytotoxic effects of DDP,whereas a necroptosis inhibitor partially reversed the cell viability loss induced by DDP.Furthermore,inhibiting autophagy enhanced both apoptosis and necroptosis induced by DDP.CONCLUSION CCT6A negatively modulates autophagy,apoptosis,and necroptosis,and CCT6A confers resistance to DDP therapy in CRC,suggesting its potential as a therapeutic target.展开更多
The complex of Eu（IH） with 1-（6-hydroxy- 1-naphthyl）- 1,3-butanedione （HNBD） was prepared for the first time and characterized by elemental analysis, IR, UV, fluorescence spectrum, and DTA-TG-DTG techniques. The...The complex of Eu（IH） with 1-（6-hydroxy- 1-naphthyl）- 1,3-butanedione （HNBD） was prepared for the first time and characterized by elemental analysis, IR, UV, fluorescence spectrum, and DTA-TG-DTG techniques. The IR and UV-visible spectra showed that Eu（Ⅲ） ion was coordinated to the HNBD ligand. The fluorescence spectrum showed the presence of Eu^3＋ characteristic emission. The TG-DTA-DTG curves showed that the thermal decomposition of the anhydrous complex was a two-stage process and the final residue was Eu2O3. The thermal decomposition kinetic parameters of the complex were evaluated from TG-DTG data by using three kinds of integral methods （Coat-Redfem equation, Horowitz and Metzger equation, Madhusudanan-Krishnan-Ninan equation）. The kinetic parameters of the first stage are E^＊ = 164.02 kJ.moll, A = 1.31 × 10^15 s^-l, AS^＊= 42.27 J·K^-l·mol^-l, △H^＊ = 159.51 kJ·mol^-l, △G^＊= 136.54 kJ·mol^-l, and n = 3.1, those of the second stage are E^＊= 128.52 kJ·mol^-l, A = 1.44× 106 s^-1, △S^＊= - 136.89 J·K^-l·mol^-l, △H^＊ = 120.41 kJ·mol^-l, △G^＊= 283.85 kJ·mol^-l, and n = 1.1.展开更多
基金Supported by the Hubei Natural Science Foundation Joint Fund Project:2023AFD139,2023AFD140the "Shizhen Young Talent" Training Program of the College of Acupuncture and Orthopedics,Hubei University of Chinese Medicinethe National Famous Traditional Chinese Medicine Expert Inheritance Studio Construction Project。
文摘Objective To investigate the effects of electroacupuncture(EA)on the expression and phosphorylation of insulin signal transduction-related proteins in the hypothalamus of insulin resistance(IR)rats.Methods There were totally seventy-five Wistar rats.Ten rats were randomly assigned to the Normal group(N).The remaining 65 rats were fed a high-fat diet,fifty successfully modeled rats were randomly divided into the Model(M),EA,Sham EA(S+EA),l-leucine(L),and l-leucine+EA(L+EA)groups,with 10 rats in each group.EA was applied at acupoints“Guanyuan(CV 4)”,“Zhongwan(CV 12)”,“Zusanli(ST 36)”,and“Fenglong(ST 40)”,with each session lasting 10 min,three times per week for 8 weeks.The S+EA group received needle insertion to a depth of≤2 mm without electrical stimulation,with the same treatment duration and same acupoint selection.Body weight,fasting blood glucose(FBG),and insulin sensitivity(glucose infusion rate,GIR)were measured.Western blot analysis was used to assess insulin receptor substrate-1(IRS-1),(Protein kinase B)Akt,glycogen synthase kinase-3β(GSK-3β),mechanistic target of rapamycin complex 1(mTORC1),and Ribosomal S6 kinase 1(S6K1),along with their phosphorylated forms.PCR was used to evaluate mRNA expression of IRS-1,Akt,and GSK-3β.Immunofluorescence was used to detect hypothalamic Akt localization.Results(1)Compared to the N group,the M group exhibited increased body weight,FBG,and phosphorylation of GSK-3β,mTORC1,and S6K1,with decreased GIR,IRS-1,Akt phosphorylation,and mRNA expression(P<0.05,P<0.01).(2)Compared to the M group,the EA and S+EA groups showed reduced body weight,FBG,GSK-3β,mTORC1,and S6K1 phosphorylation,with increased GIR,IRS-1,Akt phosphorylation,and mRNA expression(P<0.05,P<0.01).(3)Compared to the EA group,the S+EA group had higher body weight,GSK-3βphosphorylation,and mRNA expression,with reduced p-IRS-1 and p-Akt expression(P<0.05);the L and L+EA groups showed increased GSK-3β,mTORC1,and S6K1 phosphorylation,with decreased GIR,IRS-1,and Akt mRNA expression(P<0.05).(4)Compared to the L+EA group,the L group exhibited higher GSK-3β,mTORC1,and S6K1 phosphorylation,with lower GIR,Akt mRNA,and p-Akt expression(P<0.05,P<0.01).Conclusion EA positively influences body weight,glucose-lipid metabolism,and insulin sensitivity in IR rats,with regulatory effects on central insulin signal transduction-related proteins,potentially linked to its suppression of hypothalamic mTORC1/S6K1 pathway activity.
基金Supported by Shandong Provincial Natural Science Foundation,No.ZR2023MH329Project of Shandong Province Higher Educational Youth Innovation Science and Technology Program,No.2023KJ263and Natural Science Foundation of Gansu Province,China,No.22JR5RA953.
文摘BACKGROUND As a member of the chaperonin-containing tailless complex polypeptide 1(TCP1)complex,which plays a pivotal role in ensuring the accurate folding of numerous proteins,chaperonin-containing TCP1 subunit 6A(CCT6A)participates in various physiological and pathological processes.However,its effects on cell death and cancer therapy and the underlying mechanisms need further exploration in colorectal cancer(CRC)cells.AIM To explore the effects of CCT6A on cell death and cancer therapy and the underlying mechanisms in CRC.METHODS Cell proliferation was evaluated using the MTS assay,EdU staining,and colony growth assays.The expression of CCT6A was monitored by immunoblotting and quantitative PCR.CCT6A was knocked out by CRISPR-Cas9,and overexpressed by transfecting plasmids.Autophagy was examined by immunoblotting and the mCherry-GFP-LC3 assay.To monitor apoptosis and necroptosis,immunoblotting,co-immunoprecipitation,and flow cytometry were employed.RESULTS Cisplatin(DDP)exerted cytotoxic effects on CRC cells while simultaneously downregulating the expression of CCT6A.Depletion of CCT6A amplified the cytotoxic effects of DDP,whereas overexpression of CCT6A attenuated these adverse effects.CCT6A suppressed autophagy,apoptosis,and necroptosis under both basal and DDP-treated conditions.Autophagy inhibitors significantly enhanced the cytotoxic effects of DDP,whereas a necroptosis inhibitor partially reversed the cell viability loss induced by DDP.Furthermore,inhibiting autophagy enhanced both apoptosis and necroptosis induced by DDP.CONCLUSION CCT6A negatively modulates autophagy,apoptosis,and necroptosis,and CCT6A confers resistance to DDP therapy in CRC,suggesting its potential as a therapeutic target.
基金financially supported by the Important Foundation of the Educational Commission of Hubei Province (No. Z200622001)the Natural Science Foundation of the Educational Commission of Hubei Province, China (No. J200522002)
文摘The complex of Eu（IH） with 1-（6-hydroxy- 1-naphthyl）- 1,3-butanedione （HNBD） was prepared for the first time and characterized by elemental analysis, IR, UV, fluorescence spectrum, and DTA-TG-DTG techniques. The IR and UV-visible spectra showed that Eu（Ⅲ） ion was coordinated to the HNBD ligand. The fluorescence spectrum showed the presence of Eu^3＋ characteristic emission. The TG-DTA-DTG curves showed that the thermal decomposition of the anhydrous complex was a two-stage process and the final residue was Eu2O3. The thermal decomposition kinetic parameters of the complex were evaluated from TG-DTG data by using three kinds of integral methods （Coat-Redfem equation, Horowitz and Metzger equation, Madhusudanan-Krishnan-Ninan equation）. The kinetic parameters of the first stage are E^＊ = 164.02 kJ.moll, A = 1.31 × 10^15 s^-l, AS^＊= 42.27 J·K^-l·mol^-l, △H^＊ = 159.51 kJ·mol^-l, △G^＊= 136.54 kJ·mol^-l, and n = 3.1, those of the second stage are E^＊= 128.52 kJ·mol^-l, A = 1.44× 106 s^-1, △S^＊= - 136.89 J·K^-l·mol^-l, △H^＊ = 120.41 kJ·mol^-l, △G^＊= 283.85 kJ·mol^-l, and n = 1.1.
