Objective:Anshen Dingzhi prescription(ADP)is an effective remedy for treating post-traumatic stress disorder(PTSD);however,the mechanism underlying its beneficial effects is unclear.This study explores the roles of th...Objective:Anshen Dingzhi prescription(ADP)is an effective remedy for treating post-traumatic stress disorder(PTSD);however,the mechanism underlying its beneficial effects is unclear.This study explores the roles of the neuroinflammation regulated by the FKBP prolyl isomerase 5(FKBP5)-IκB kinase alpha(IKKα)-nuclear factor kappa-B(NF-κB)-NOD-like receptor thermal protein domain-associated protein 3(NLRP3)signaling pathway in PTSD.Methods:The primary components of ADP,including ginsenosides Rg1 and Rb1,were quantified using ultra-performance liquid chromatography.Twelve C57BL/6 mice were allocated to control(D0)and experimental groups on days one,seven,and 14 of single prolonged stress(SPS).Eighteen C57BL/6 mice were allocated to control,SPS,and MCC950,an NLRP3 inhibitor(5 mg/kg)groups.Finally,24 C57BL/6 mice were allocated to control,SPS,paroxetine hydrochloride(PRX),or ADP(18.4 and 36.8 mg/kg)groups.Mice were administered MCC950,PRX,or ADP for 14 days.The open field test and elevated plus maze were used to evaluate anxiety-like behaviors,whereas fear memory extinction was evaluated using the fear memory test.Western blotting was employed to evaluate the expression levels of the FKBP5-IKKα-NF-κB-NLRP3 signaling pathway,tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1β.The expression of FKBP5 and NLRP3 was further confirmed by immunofluorescence staining.Results:The amounts of ginsenosides Rg1 and Rb1 in ADP were(96.85±1.14)and(9.04±0.22)μg/g,respectively.Compared with the D0 group,the levels of the inflammatory cytokine proteins,TNF-α,IL-6,and IL-1β were elevated 1.33-to 1.51-fold and those of FKBP5-IKKα-NF-κB-NLRP3 signaling pathway were increased 1.16-to 1.41-fold in the hippocampus of the D14 group(P<0.05);the fluorescence intensity of FKBP5 and NLRP3 was also markedly increased(1.33-1.79-fold)in the hippocampus of the D14 group(P<0.5).Notably,injection of MCC950(5 mg/kg)reduced the levels of FKBP5-IKKα-NF-κB-NLRP3(0.80-0.88-fold)and inflammatory cytokines(0.74-0.83-fold),thereby improving the PTSD-like behaviors induced by SPS(P<0.05).In addition,ADP(36.8 g/kg)significantly improved PTSD-like behaviors and reduced levels of hippocampal inflammatory cytokines(0.70-0.79-fold)and FKBP5-IKKα-NF-κB-NLRP3(0.50-0.79-fold)(P<0.05)in SPS mice.Conclusion:The results suggest a potential therapeutic benefit of ADP in PTSD due to the inhibition of the FKBP5-IKKα-NF-κBNLRP3 signaling pathway.展开更多
基金supported by the National Natural Science Foundation of China(82404995,82404890)Research Funds of Center for Xin’an Medicine and Modernization of Traditional Chinese Medicine of IHM(2023CXMMTCM013)+3 种基金Scientific Research Program of Anhui Provincial Department of Education(2024AH051036,2024AH040137,2024AH051044)Excellent Funding for Academic and Scientific Research Activities for Academic and Technological Leaders in Anhui Province(2022D317)Key Research and Development Plan of Anhui Province(202104j07020004)Anhui University of Chinese Medicine Talent Support Program(DT2300000173).
文摘Objective:Anshen Dingzhi prescription(ADP)is an effective remedy for treating post-traumatic stress disorder(PTSD);however,the mechanism underlying its beneficial effects is unclear.This study explores the roles of the neuroinflammation regulated by the FKBP prolyl isomerase 5(FKBP5)-IκB kinase alpha(IKKα)-nuclear factor kappa-B(NF-κB)-NOD-like receptor thermal protein domain-associated protein 3(NLRP3)signaling pathway in PTSD.Methods:The primary components of ADP,including ginsenosides Rg1 and Rb1,were quantified using ultra-performance liquid chromatography.Twelve C57BL/6 mice were allocated to control(D0)and experimental groups on days one,seven,and 14 of single prolonged stress(SPS).Eighteen C57BL/6 mice were allocated to control,SPS,and MCC950,an NLRP3 inhibitor(5 mg/kg)groups.Finally,24 C57BL/6 mice were allocated to control,SPS,paroxetine hydrochloride(PRX),or ADP(18.4 and 36.8 mg/kg)groups.Mice were administered MCC950,PRX,or ADP for 14 days.The open field test and elevated plus maze were used to evaluate anxiety-like behaviors,whereas fear memory extinction was evaluated using the fear memory test.Western blotting was employed to evaluate the expression levels of the FKBP5-IKKα-NF-κB-NLRP3 signaling pathway,tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1β.The expression of FKBP5 and NLRP3 was further confirmed by immunofluorescence staining.Results:The amounts of ginsenosides Rg1 and Rb1 in ADP were(96.85±1.14)and(9.04±0.22)μg/g,respectively.Compared with the D0 group,the levels of the inflammatory cytokine proteins,TNF-α,IL-6,and IL-1β were elevated 1.33-to 1.51-fold and those of FKBP5-IKKα-NF-κB-NLRP3 signaling pathway were increased 1.16-to 1.41-fold in the hippocampus of the D14 group(P<0.05);the fluorescence intensity of FKBP5 and NLRP3 was also markedly increased(1.33-1.79-fold)in the hippocampus of the D14 group(P<0.5).Notably,injection of MCC950(5 mg/kg)reduced the levels of FKBP5-IKKα-NF-κB-NLRP3(0.80-0.88-fold)and inflammatory cytokines(0.74-0.83-fold),thereby improving the PTSD-like behaviors induced by SPS(P<0.05).In addition,ADP(36.8 g/kg)significantly improved PTSD-like behaviors and reduced levels of hippocampal inflammatory cytokines(0.70-0.79-fold)and FKBP5-IKKα-NF-κB-NLRP3(0.50-0.79-fold)(P<0.05)in SPS mice.Conclusion:The results suggest a potential therapeutic benefit of ADP in PTSD due to the inhibition of the FKBP5-IKKα-NF-κBNLRP3 signaling pathway.
