Objective:This article uses the method of network pharmacology to study the related mechanism of Guilu Erxian Gum in the treatment of osteoporosis.Methods:Based on the TCMSP database,ETCM database,chemistry database,a...Objective:This article uses the method of network pharmacology to study the related mechanism of Guilu Erxian Gum in the treatment of osteoporosis.Methods:Based on the TCMSP database,ETCM database,chemistry database,and DrugBank database,the potential active ingredients and related targets of Guilu Erxian Gum were obtained.The known therapeutic targets of osteoporosis were obtained from OMIM and Genecards databases,and the STRING database was used.A protein interaction network(PPI)of active ingredients-disease targets was established,and the topological parameters of the network were analyzed using Cytoscape 3.8.0 software to obtain key active ingredients and their targets.In R4.0.2,the Bioconductor data package was used to analyze the GO biological function and KEGG pathway analysis of key targets,and obtain the effective ingredients and targets of Guilu Erxian Gum for treating osteoporosis.Results:The prediction results show that Guilu Erxian Gum has 87 active ingredients and 2305 targets,and there are 4141 known therapeutic targets for osteoporosis.The two act together to obtain a total of 71 PPI core genes.The GO biological process analysis yielded 95 entries,and the KEGG pathway analysis yielded 115 pathways.Conclusion:The analysis results show that Guilu Erxian Gum may play an anti-osteoporosis effect by regulating inflammatory factors,promoting osteoblast differentiation,inhibiting osteoclast formation,and improving microcirculation.The pathways involved include TNF signaling pathways,IL-17 signaling pathway,NF-κB signaling pathway,HIF-1 signaling pathway,MAPK signaling pathway,PI3K-Akt signaling pathway,etc.This study provides a theoretical basis for further elucidating the pharmacological mechanism of Guilu Erxian Gum in the treatment of osteoporosis.展开更多
Objective To investigate the intervention effect and mechanism of Guilu Erxian Glue(GEG)on fear memory extinction in a rat model of post-traumatic stress disorder(PTSD).Methods SD rats were randomly divided into contr...Objective To investigate the intervention effect and mechanism of Guilu Erxian Glue(GEG)on fear memory extinction in a rat model of post-traumatic stress disorder(PTSD).Methods SD rats were randomly divided into control group,model group,GEG group(3.6 g/kg),and paroxetine group(10 mg/kg),with 10 rats in each group.A 6-day continuous inescapable foot shock was used as the stressor to establish a PTSD rat model.展开更多
Background: Myelosuppression is a common chemotherapy-related toxicity and hematopoietic stem cells (HSCs)senescence is the main mechanism. Guilu Erxian Glue (GEG) is an effective Chinese herbal prescription for the t...Background: Myelosuppression is a common chemotherapy-related toxicity and hematopoietic stem cells (HSCs)senescence is the main mechanism. Guilu Erxian Glue (GEG) is an effective Chinese herbal prescription for the treatment of myelosuppression in clinical, the potential mechanism needs further investigation.Objective: To explore the protective effect of GEG on HSCs during chemotherapy-induced myelosuppression and its mechanism.Methods: Eighteen H22 hepatoma-bearing mice were divided equally into three groups: control, cisplatin (DDP),and DDP+GEG groups. Peripheral blood cell counts and bone marrow nucleated cell counts were performed.Bone marrow histopathology was assessed by hematoxylin and eosin (H&E) staining. DNA damage was assessed by γ-H2AX immunofluorescence staining. The serum containing GEG was prepared for in vitro experiments. Superoxide dismutase (SOD) activity, cell viability, cell cycle, and aging cells were detected in vivo and in vitro. In addition, western blot, quantitative reverse transcription polymerase chain reaction (qRT-PCR), and chromatin immunoprecipitation (CHIP) assay were applied to explore the molecular mechanism.Results: GEG administration resulted in a significant increase in peripheral blood cells and bone marrow nucleated cells (P < 0.01), and improved the pathological histomorphology of bone marrow. Treatment with GEG or silencing of the p16Ink4a gene enhanced the SOD activity and viability of HSCs, while reducing DDP-induced DNA damage, G2-M cell cycle arrest and senescent cell formation (P < 0.001). GEG decreased both mRNA and protein levels of p16, while increased the protein and mRNA expression levels of CDK4, CDK6, and pRb both in vivo and in vitro experiments (P < 0.05). ChIP-qPCR showed that the p16Ink4apromoter region was enriched with JMJD2and H3K36me2/me3 protein. GEG treatment up-regulated the protein and mRNA expression levels of JMJD2 and simultaneously down-regulated H3K36me2/me3 protein levels.Conclusion: Our findings suggest that GEG exerts regulatory effects on p16ink4a gene expression through modulating the levels of H3K36 methylation. This study provides important mechanistic insights into the effects of GEG on HSCs and suggests potential therapeutic applications for GEG in myelosuppression.展开更多
Objective To evaluate the effect of Guilu Erxian Glue(龟鹿二仙胶,GEG)on cyclophosphamide(CTX)-induced bone marrow hematopoietic stem cells(HSCs)senescence in mice and explore the underlying mechanism.Methods The H22 l...Objective To evaluate the effect of Guilu Erxian Glue(龟鹿二仙胶,GEG)on cyclophosphamide(CTX)-induced bone marrow hematopoietic stem cells(HSCs)senescence in mice and explore the underlying mechanism.Methods The H22 liver cancer ascites lump model was established in male Kunming mice by injecting intraperitoneally(i.p.)with 5×10^6/mL H22 cells per mouse.Fifty tumor-bearing mice were divided into the control,model,pifithrin-α,GEG,and GEG+pifithrin-αgroups using a random number table,10 mice in each group.CTX(100 mg/kg i.p.)was administrated to mice from day 1 to day 3(d1–d3)continuously except for the control group.The mice in the pifithrin-α,GEG and GEG+pifithrin-αgroups were treated with pifithrin-α(2.2 mg/(kg·d)i.p.)for 6 consecutive days(d4–d9),GEG(9.5 g/(kg·d)i.p.)for 9 consecutive days(d1–d9),and GEG plus pifithrin-α,respectively.HSCs were collected after 9-d drug treatment.The anti-aging effect of GEG was studied by cell viability,cell cycle,andβ-galactosidase(β-gal)assays.The mRNA and protein expressions of cyclin-dependent kinase 2(CDK2),CDK4,inhibitor of cyclin-dependent kinase 4a encoding the tumor suppressor protein p16^(p16^INK4a),p21^Cip1/Waf1,p53,and phosphorylated retinoblastoma(pRb)were evaluated by quantitative real-time reverse transcription-polymerase chain reaction and semi-quantitative Western blot,respectively.Results Compared with the model group,GEG increased cell viability as well as proliferation(P<0.05 or P<0.01)and reducedβ-gal expression.Furthermore,GEG significantly decreased the expressions of p16^INK4a,p53 and p21^Cip1/Waf1 proteins,and increased the expressions of CDK2,CDK4 and pRb proteins compared with the model group(P<0.05 or P<0.01).Conclusion GEG can alleviate CTX-induced HSCs senescence in mice,and the p16^INK4a-Rb signaling pathway might be the underlying mechanism.展开更多
基金National Natural Science Foundation of China(No.81973719)Science and technology research project of Liaoning Provincial Department of Education(No.L201921)Liaoning Provincial Science And Technology Plan Project(No.2020JH2/10300068)。
文摘Objective:This article uses the method of network pharmacology to study the related mechanism of Guilu Erxian Gum in the treatment of osteoporosis.Methods:Based on the TCMSP database,ETCM database,chemistry database,and DrugBank database,the potential active ingredients and related targets of Guilu Erxian Gum were obtained.The known therapeutic targets of osteoporosis were obtained from OMIM and Genecards databases,and the STRING database was used.A protein interaction network(PPI)of active ingredients-disease targets was established,and the topological parameters of the network were analyzed using Cytoscape 3.8.0 software to obtain key active ingredients and their targets.In R4.0.2,the Bioconductor data package was used to analyze the GO biological function and KEGG pathway analysis of key targets,and obtain the effective ingredients and targets of Guilu Erxian Gum for treating osteoporosis.Results:The prediction results show that Guilu Erxian Gum has 87 active ingredients and 2305 targets,and there are 4141 known therapeutic targets for osteoporosis.The two act together to obtain a total of 71 PPI core genes.The GO biological process analysis yielded 95 entries,and the KEGG pathway analysis yielded 115 pathways.Conclusion:The analysis results show that Guilu Erxian Gum may play an anti-osteoporosis effect by regulating inflammatory factors,promoting osteoblast differentiation,inhibiting osteoclast formation,and improving microcirculation.The pathways involved include TNF signaling pathways,IL-17 signaling pathway,NF-κB signaling pathway,HIF-1 signaling pathway,MAPK signaling pathway,PI3K-Akt signaling pathway,etc.This study provides a theoretical basis for further elucidating the pharmacological mechanism of Guilu Erxian Gum in the treatment of osteoporosis.
文摘Objective To investigate the intervention effect and mechanism of Guilu Erxian Glue(GEG)on fear memory extinction in a rat model of post-traumatic stress disorder(PTSD).Methods SD rats were randomly divided into control group,model group,GEG group(3.6 g/kg),and paroxetine group(10 mg/kg),with 10 rats in each group.A 6-day continuous inescapable foot shock was used as the stressor to establish a PTSD rat model.
基金supported by the Natural Science Foundation of Zhejiang Province of China (No.LY24H270010)the National Natural Science Foundation of China (No.81904197)。
文摘Background: Myelosuppression is a common chemotherapy-related toxicity and hematopoietic stem cells (HSCs)senescence is the main mechanism. Guilu Erxian Glue (GEG) is an effective Chinese herbal prescription for the treatment of myelosuppression in clinical, the potential mechanism needs further investigation.Objective: To explore the protective effect of GEG on HSCs during chemotherapy-induced myelosuppression and its mechanism.Methods: Eighteen H22 hepatoma-bearing mice were divided equally into three groups: control, cisplatin (DDP),and DDP+GEG groups. Peripheral blood cell counts and bone marrow nucleated cell counts were performed.Bone marrow histopathology was assessed by hematoxylin and eosin (H&E) staining. DNA damage was assessed by γ-H2AX immunofluorescence staining. The serum containing GEG was prepared for in vitro experiments. Superoxide dismutase (SOD) activity, cell viability, cell cycle, and aging cells were detected in vivo and in vitro. In addition, western blot, quantitative reverse transcription polymerase chain reaction (qRT-PCR), and chromatin immunoprecipitation (CHIP) assay were applied to explore the molecular mechanism.Results: GEG administration resulted in a significant increase in peripheral blood cells and bone marrow nucleated cells (P < 0.01), and improved the pathological histomorphology of bone marrow. Treatment with GEG or silencing of the p16Ink4a gene enhanced the SOD activity and viability of HSCs, while reducing DDP-induced DNA damage, G2-M cell cycle arrest and senescent cell formation (P < 0.001). GEG decreased both mRNA and protein levels of p16, while increased the protein and mRNA expression levels of CDK4, CDK6, and pRb both in vivo and in vitro experiments (P < 0.05). ChIP-qPCR showed that the p16Ink4apromoter region was enriched with JMJD2and H3K36me2/me3 protein. GEG treatment up-regulated the protein and mRNA expression levels of JMJD2 and simultaneously down-regulated H3K36me2/me3 protein levels.Conclusion: Our findings suggest that GEG exerts regulatory effects on p16ink4a gene expression through modulating the levels of H3K36 methylation. This study provides important mechanistic insights into the effects of GEG on HSCs and suggests potential therapeutic applications for GEG in myelosuppression.
基金Supported by the National Natural Science Foundation of 6hina(No.81904197)Natural Science Foundation of Zhejiang Province(No.LQ15H290002)and 2019 Research and Innovation Fund Project for Young and Middle-aged Researchers of Zhejiang Chinese Medical University(No.KC201944)。
文摘Objective To evaluate the effect of Guilu Erxian Glue(龟鹿二仙胶,GEG)on cyclophosphamide(CTX)-induced bone marrow hematopoietic stem cells(HSCs)senescence in mice and explore the underlying mechanism.Methods The H22 liver cancer ascites lump model was established in male Kunming mice by injecting intraperitoneally(i.p.)with 5×10^6/mL H22 cells per mouse.Fifty tumor-bearing mice were divided into the control,model,pifithrin-α,GEG,and GEG+pifithrin-αgroups using a random number table,10 mice in each group.CTX(100 mg/kg i.p.)was administrated to mice from day 1 to day 3(d1–d3)continuously except for the control group.The mice in the pifithrin-α,GEG and GEG+pifithrin-αgroups were treated with pifithrin-α(2.2 mg/(kg·d)i.p.)for 6 consecutive days(d4–d9),GEG(9.5 g/(kg·d)i.p.)for 9 consecutive days(d1–d9),and GEG plus pifithrin-α,respectively.HSCs were collected after 9-d drug treatment.The anti-aging effect of GEG was studied by cell viability,cell cycle,andβ-galactosidase(β-gal)assays.The mRNA and protein expressions of cyclin-dependent kinase 2(CDK2),CDK4,inhibitor of cyclin-dependent kinase 4a encoding the tumor suppressor protein p16^(p16^INK4a),p21^Cip1/Waf1,p53,and phosphorylated retinoblastoma(pRb)were evaluated by quantitative real-time reverse transcription-polymerase chain reaction and semi-quantitative Western blot,respectively.Results Compared with the model group,GEG increased cell viability as well as proliferation(P<0.05 or P<0.01)and reducedβ-gal expression.Furthermore,GEG significantly decreased the expressions of p16^INK4a,p53 and p21^Cip1/Waf1 proteins,and increased the expressions of CDK2,CDK4 and pRb proteins compared with the model group(P<0.05 or P<0.01).Conclusion GEG can alleviate CTX-induced HSCs senescence in mice,and the p16^INK4a-Rb signaling pathway might be the underlying mechanism.
