The title compound of physalin B（C28H32O9）, a main active physalin of Physalis angulata L, was isolated from the whole plant of Physalis angulata L, and characterized by X-ray diffraction analysis. It crystallizes i...The title compound of physalin B（C28H32O9）, a main active physalin of Physalis angulata L, was isolated from the whole plant of Physalis angulata L, and characterized by X-ray diffraction analysis. It crystallizes in the monoclinic system, space group P21 with C28H32O9, a = 12.4996（2）, b = 14.35620（10）, c = 14.75190（10）, V = 2607.97（5） 3, Z = 4, Dc = 1.382 mg/cm3, Mr = 542.56, F（000） = 1152, and μ = 0.870 mm-1. The final R = 0.0389 and wR = 0.1037 for 47670 observed reflections（I 〉 2σ（I））. The rigid molecule consists of eight fused rings involving two lactones. There are two C28H32O9 molecules in an symmetric unit, and the title compound is stacked into a 3D layer structure through hydrogen bonds. In the 5~20 μmol/L range, physalin B can significantly inhibit the secretion of inflammatory cytokines TNF-α and IL-6 on RAW264.7 cells. The results suggest that physalin B has anti-inflammatory activity in vitro.展开更多
OBJECTIVE Extracellular amyloid-β(Aβ) plaques are one of the major pathological hallmarks of Alzheimer disease(AD). Therefore, decreasing Aβ levels is one strategyfor preventing the etiology of AD. Aβ peptides are...OBJECTIVE Extracellular amyloid-β(Aβ) plaques are one of the major pathological hallmarks of Alzheimer disease(AD). Therefore, decreasing Aβ levels is one strategyfor preventing the etiology of AD. Aβ peptides are generated from the cleavage of amyloid precursor protein(APP) by the β-secretase(BACE1) and γ-secretase(PS1). Inhibition of these secretases represents an obvious logical strategy to inhibit the generation of Aβ. In addition, signal transducer and activator of transcription 3(STAT3) is known to regulate many genes, and to significantly affect Aβ generation by controlling BACE1 and PS1 expression. Physalin B(PB), one of the major active steroidal constituents of solanaceaephysalis plants, possesses a wide variety of biological activities. PB down-regulates BACE1 and PS1 expression while it is unclear whether PB can regulate Aβ in N2 a/APPswe cells, and if so, whether it is by inhibiting the phosphorylation of STAT3. METHODS N2 a/APPswe cells were treated with PB in different concentrations for 24 h.(1) We used CCK8 method to detect the effects of different concentrations of PB on cell viability, and selected the best concentration for drug treatment to the cells.(2)The contents of Aβ40 and Aβ42 were determined by ELISA.(3) Western blotting was used to detect the expression levels of p-STAT3 and APP metabolism-related proteins, including APP, CTFα, CTFβ, BACE1,PS1, ADAM10 and so on.(4) RT-PCR was performed to detected the m RNA expression of BACE1 and PS1.(5) β-secretase activity Fluorometric assay kit was used to analyzed β-secretase activity.(6) In order to further explore the underlying mechanisms, N2 a/APPswe cells were pre-treated with 100 μmol·L-1 S3 I-201(a STAT3 inhibitor,can effectively prevent STAT3 phosphorylation) for 30 min and then treated with 3 μmol·L-1 PB incubated for 24 h. Then we evaluated the level of expression of STAT3 and p-STAT3 by Western blotting. RESULTS(1) CCK8 experiment results illustrated that PB did not show cytotoxicity at the applied concentration when cells were treated with PB(0, 0.3, 1 and 3 μmol·L-1). So, we used these concentrations in the following experiment.(2) ELISA results showed, compared to the control group, the contents of Aβ40 and Aβ42 decreased with the increasing of PB concentration.(3)According to Western blotting results, PB significant down-regulated the expression of BACE1, PS1, APP, CTFβ and p-STAT3 compared to the control group.(4) RT-PCR results indicated that PB can reduced the m RNA expression of BACE1 and PS1 effectively.(5) It turns out that PB can significantly inhibit BACE1 activity tested by BACE1 activity Fluorometric assay kit.(6) S3 I-201 had a similar manner to significantly inhibited STAT3 phosphorylation with PB through Western blotting. Moreover, co-treated with S3 I-201 and PB inhibited STAT3 phosphorylation much more than treatment with S3 I-201 alone. CONCLUSION These findings indicate that PB can effectively inhibit the expression of BACE1 and PS1 to reduce Aβ secretion by inhibiting the phosphorylation of STAT3.展开更多
Physalin B(PB),one of the major active steroidal constituents of Solanaceae Physalis plants,has a wide variety of biological activities.We found that PB significantly down-regulatedβ-amyloid(Aβ)secretion in N2a/APPs...Physalin B(PB),one of the major active steroidal constituents of Solanaceae Physalis plants,has a wide variety of biological activities.We found that PB significantly down-regulatedβ-amyloid(Aβ)secretion in N2a/APPsw cells.However,the underlying mechanisms are not well understood.In the current study,we investigated the changes in key enzymes involved inβ-amyloid precursor protein(APP)metabolism and other APP metabolites by treating N2a/APPsw cells with PB at different concentrations.The results indicated that PB reduced Aβ secretion,which was caused by down-regulation of β-secretase(BACE1)expression,as indicated at both the protein and mRNA levels.Further research revealed that PB regulated BACE1 expression by inducing the activation of forkhead box O1(FoxO1)and inhibiting the phosphorylation of signal transducer and activator of transcription 3(STAT3).In addition,the effect of PB on BACE1 expression and Aβsecretion was reversed by treatment with FoxO1 siRNA and STAT3 antagonist S3I-201.In conclusion,these data demonstrated that PB can effectively down-regulate the expression of BACE1 to reduce Aβsecretion by activating the expression of FoxO1 and inhibiting the phosphorylation of STAT3.展开更多
基金Supported by the Guangdong Provincial Science and Technology Projects(No.2011B031700072)Guangdong Natural Science Foundation(No.9151063201000036,S2013010013484)Guangdong Provincial Administration of traditional Chinese medicine project(No.2008435)
文摘The title compound of physalin B（C28H32O9）, a main active physalin of Physalis angulata L, was isolated from the whole plant of Physalis angulata L, and characterized by X-ray diffraction analysis. It crystallizes in the monoclinic system, space group P21 with C28H32O9, a = 12.4996（2）, b = 14.35620（10）, c = 14.75190（10）, V = 2607.97（5） 3, Z = 4, Dc = 1.382 mg/cm3, Mr = 542.56, F（000） = 1152, and μ = 0.870 mm-1. The final R = 0.0389 and wR = 0.1037 for 47670 observed reflections（I 〉 2σ（I））. The rigid molecule consists of eight fused rings involving two lactones. There are two C28H32O9 molecules in an symmetric unit, and the title compound is stacked into a 3D layer structure through hydrogen bonds. In the 5~20 μmol/L range, physalin B can significantly inhibit the secretion of inflammatory cytokines TNF-α and IL-6 on RAW264.7 cells. The results suggest that physalin B has anti-inflammatory activity in vitro.
基金National Natural Science Foundation(U1604108)Key Science and Technology Program of Henan Province(152102310112+1 种基金 162102310121)National Training Program of Innovation for Undergraduates(201710472009)
文摘OBJECTIVE Extracellular amyloid-β(Aβ) plaques are one of the major pathological hallmarks of Alzheimer disease(AD). Therefore, decreasing Aβ levels is one strategyfor preventing the etiology of AD. Aβ peptides are generated from the cleavage of amyloid precursor protein(APP) by the β-secretase(BACE1) and γ-secretase(PS1). Inhibition of these secretases represents an obvious logical strategy to inhibit the generation of Aβ. In addition, signal transducer and activator of transcription 3(STAT3) is known to regulate many genes, and to significantly affect Aβ generation by controlling BACE1 and PS1 expression. Physalin B(PB), one of the major active steroidal constituents of solanaceaephysalis plants, possesses a wide variety of biological activities. PB down-regulates BACE1 and PS1 expression while it is unclear whether PB can regulate Aβ in N2 a/APPswe cells, and if so, whether it is by inhibiting the phosphorylation of STAT3. METHODS N2 a/APPswe cells were treated with PB in different concentrations for 24 h.(1) We used CCK8 method to detect the effects of different concentrations of PB on cell viability, and selected the best concentration for drug treatment to the cells.(2)The contents of Aβ40 and Aβ42 were determined by ELISA.(3) Western blotting was used to detect the expression levels of p-STAT3 and APP metabolism-related proteins, including APP, CTFα, CTFβ, BACE1,PS1, ADAM10 and so on.(4) RT-PCR was performed to detected the m RNA expression of BACE1 and PS1.(5) β-secretase activity Fluorometric assay kit was used to analyzed β-secretase activity.(6) In order to further explore the underlying mechanisms, N2 a/APPswe cells were pre-treated with 100 μmol·L-1 S3 I-201(a STAT3 inhibitor,can effectively prevent STAT3 phosphorylation) for 30 min and then treated with 3 μmol·L-1 PB incubated for 24 h. Then we evaluated the level of expression of STAT3 and p-STAT3 by Western blotting. RESULTS(1) CCK8 experiment results illustrated that PB did not show cytotoxicity at the applied concentration when cells were treated with PB(0, 0.3, 1 and 3 μmol·L-1). So, we used these concentrations in the following experiment.(2) ELISA results showed, compared to the control group, the contents of Aβ40 and Aβ42 decreased with the increasing of PB concentration.(3)According to Western blotting results, PB significant down-regulated the expression of BACE1, PS1, APP, CTFβ and p-STAT3 compared to the control group.(4) RT-PCR results indicated that PB can reduced the m RNA expression of BACE1 and PS1 effectively.(5) It turns out that PB can significantly inhibit BACE1 activity tested by BACE1 activity Fluorometric assay kit.(6) S3 I-201 had a similar manner to significantly inhibited STAT3 phosphorylation with PB through Western blotting. Moreover, co-treated with S3 I-201 and PB inhibited STAT3 phosphorylation much more than treatment with S3 I-201 alone. CONCLUSION These findings indicate that PB can effectively inhibit the expression of BACE1 and PS1 to reduce Aβ secretion by inhibiting the phosphorylation of STAT3.
基金supported by the National Natural Science Foundation(No.U1604108)Key Science and Technology Program of Henan Province(Nos.202102310020 and 182102311148)+1 种基金the Program for Innovative Research Team(in Science and Technology)in University of Henan Province(No.20IRTSTHN030)the Graduate Scientific Research Innovation Support Project of Xinxiang Medical University(No.YJSCX201866Y).
文摘Physalin B(PB),one of the major active steroidal constituents of Solanaceae Physalis plants,has a wide variety of biological activities.We found that PB significantly down-regulatedβ-amyloid(Aβ)secretion in N2a/APPsw cells.However,the underlying mechanisms are not well understood.In the current study,we investigated the changes in key enzymes involved inβ-amyloid precursor protein(APP)metabolism and other APP metabolites by treating N2a/APPsw cells with PB at different concentrations.The results indicated that PB reduced Aβ secretion,which was caused by down-regulation of β-secretase(BACE1)expression,as indicated at both the protein and mRNA levels.Further research revealed that PB regulated BACE1 expression by inducing the activation of forkhead box O1(FoxO1)and inhibiting the phosphorylation of signal transducer and activator of transcription 3(STAT3).In addition,the effect of PB on BACE1 expression and Aβsecretion was reversed by treatment with FoxO1 siRNA and STAT3 antagonist S3I-201.In conclusion,these data demonstrated that PB can effectively down-regulate the expression of BACE1 to reduce Aβsecretion by activating the expression of FoxO1 and inhibiting the phosphorylation of STAT3.
