目的:基于网络药理学及分子对接技术,探究三棱-莪术药对在辅助治疗卵巢癌中的作用机制。方法:活性成分及其作用靶点收集自中药系统药理学分析平台,卵巢癌相关靶点收集自Genecard数据库。上述靶点取交集后,结合STRING数据库及Cytoscape...目的:基于网络药理学及分子对接技术,探究三棱-莪术药对在辅助治疗卵巢癌中的作用机制。方法:活性成分及其作用靶点收集自中药系统药理学分析平台,卵巢癌相关靶点收集自Genecard数据库。上述靶点取交集后,结合STRING数据库及Cytoscape软件构建PPI网络。应用R及Cytoscape软件整合构建中药-活性成分-疾病-靶点网络。同时,应用R软件进行基因本体论(gene ontology,GO)功能富集、京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集,并通过Cytoscape/CluGO进行KEGG通路聚类分析。综合分析各网络,获取关键靶点及其对应活性成分后应用Autodock vina及Pymol等软件进行分子对接及可视化研究。结果:本研究在三棱-莪术药对中,共获得8个活性成分,164个作用靶点,Genecards数据库获得1540个符合条件的卵巢癌相关靶点。取交集获得27个三棱-莪术药对治疗卵巢癌的潜在靶点。分析PPI网络并获取关键靶点为CASP3、ESR1、JUN、PTGS2、AR、MAPK14、PGR、CASP8、CASP9、CAS及PPARG。GO及KEGG分析表明相关作用机制可能涉及凋亡、白细胞介素17(interleukin-17,IL-17)信号通路、乙肝信号通路、雌激素信号通路、甲状腺激素信号通路,卵巢类固醇等多条信号通路。分子对接表明核心成分芒柄花黄素、β-谷甾醇、常春藤皂苷元与关键靶点结合性良好,其中各活性成分与PTGS2结合最佳,结合能均小于-9.0 kcal/mol。结论:该研究初步揭示了三棱-莪术药对治疗卵巢癌的药理机制,这为后期探究该药对及其所在复方治疗卵巢癌提供了较为系统的依据。展开更多
OBJECTIVE: To study the anti-tumor effects of the extracts from Huangqi(Radix Astragali Mongolici)and Ezhu(Rhizoma Curcumae Phaeocaulis) on the growth of Lewis lung carcinoma(LLC) in a xenograft mouse model and to inv...OBJECTIVE: To study the anti-tumor effects of the extracts from Huangqi(Radix Astragali Mongolici)and Ezhu(Rhizoma Curcumae Phaeocaulis) on the growth of Lewis lung carcinoma(LLC) in a xenograft mouse model and to investigate the possible underlying mechanism.METHODS: LLC tumor-bearing C57 BL/6 mice were treated with normal saline, cisplatin(2 mg/kg intraperitoneally every other day), or Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis)(1∶1, 2∶1, or 3∶1 ratio;5, 8, or 11 g/kg crude drug intragastrically every day) for 15 d.Body weights and tumor volumes were measured every other day. Tumors were excised on day 15 and analyzed. Tumor microvessel density(MVD)was assessed by immunohistochemical staining of CD34;and expression of vascular endothelial cell growth factor(VEGF), the mitogen-activated protein kinases p38 mitogen-activated protein kinase(MAPK), extracellular signal-regulated kinases 1 and 2(ERK1/2), and Jun N-terminal kinase(JNK)and their phosphorylated forms were assessed by Western blotting.RESULTS: Treatment with cisplatin caused a significant loss of body weight compared with controls,whereas Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) extract combinations had no effect. Extracts from Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) significantly decreased tumor weight and tumor MVD compared with controls,and at the 3∶1 treatment group had similar efficacy to cisplatin in reducing MVD. Tumors from Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) treatments also showed decreased p38 MAPK, p-p38 MAPK, ERK1/2, p-ERK1/2,JNK, and p-JNK expression compared with the control group(all P < 0.01). VEGF protein expression was significantly reduced in the 2∶1 and 3∶1 treatment groups compared with the control group(P < 0.01).CONCLUSION: Extracts from Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) hindered LLC growth in the xenograft mouse model, possibly via inhibition of the MAPK signaling pathway, VEGF production, and tumor angiogenesis.展开更多
OBJECTIVE: To investigate the effect of optimal combination(E) of Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) on proliferation and apoptosis of A549 lung cancer cells and the possible mec...OBJECTIVE: To investigate the effect of optimal combination(E) of Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) on proliferation and apoptosis of A549 lung cancer cells and the possible mechanism underpinning the action.METHODS: A uniform design method was used to optimize the E of Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) in A549 lung cancer cells. MTS assay was applied to analyze the effect of the component formula ofHuangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) on A549 cells viability in various uniform design groups. A549 cells with exponential growth in routine culture were exposed to Co Cl_2(200 μmol/L) to mimic hypoxic conditions. Group 0 was treated with RPMI-1640, the group Co Cl_2 was treated with Co Cl_2(200 μmol/L),the group DDP + Co Cl_2 was treated with 4 mg/L Cisplatin injection(DDP) + Co Cl_2(200 μmol/L), and the drug group was treated with various dose of E(0.5 E, 1 E, 2 E) + Co Cl_2(200 μmol/L). All groups were cultured for 24 h. Cell apoptosis was measured by Annexin V-FITC/propidium iodide double staining and flow cytometry. Western blot assay and quantitative real-time polymerase chain reaction(q RT-PCR) were employed to detect the protein and m RNA expression of B-celllymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax) and cysteinyl aspartate specific proteinase-3(caspase-3).RESULTS: The E obtained by the uniform design was comprise of 200 mg/L Astragalus polysaccharide(X1) and 32 mg/L Curcumin(X3). Group DDP+Co Cl_2, group 1 E + Co Cl_2 and group 2 E + Co Cl_2 promoted the apoptosis of A549 cells(P < 0.05). Group1 E + Co Cl_2 and group 2 E + Co Cl_2 had no statistically significant differences compared with the group DDP + Co Cl_2(P > 0.05). Compared with group 0, various doses of E + Co Cl_2 could up-regulate the expression of Bax and caspase-3 and down-regulate the expression of Bcl-2 at protein and m RNA levels(P < 0.05).CONCLUSION: Astragalus polysaccharide and Curcumin was the optimal combination of Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Cur-cumae Phaeocaulis). E promoted the apoptosis of A549 cells. Combination of Astragalus polysaccharide and Curcumin increased the expression of Bax and caspase-3, and decreased the expression of Bcl-2 to initiate apoptosis in A549 cells under chemical-induced hypoxia.展开更多
OBJECTIVE:To investigate the efficacy of Biejia(Carapax Trionycis)and Ezhu(Rhizoma Curcumae Phaeocaulis)couplet medicine on epithelial-mesenchymal transition(EMT),invasion and migration of MDA-MB-231 triple negative b...OBJECTIVE:To investigate the efficacy of Biejia(Carapax Trionycis)and Ezhu(Rhizoma Curcumae Phaeocaulis)couplet medicine on epithelial-mesenchymal transition(EMT),invasion and migration of MDA-MB-231 triple negative breast cancer(TNBC)cells based on PI3K/Akt/mTOR signaling pathway.METHODS:MDA-MB-231 cells were treated with different medicated serum as Biejia-,Ezhu-,Biejia-Ezhu(BJ-,EZ-,BJ-EZ-)groups,intervened with no drug rat serum and paclitaxel with final concentration of 33 nM(IC50)as negative and positive control(NC and PC)groups.CCK-8 assay,scratch test,and Transwell assay were used to examine cell proliferation,invasion,and migration.The expression of E-cadherin,N-cadherin,Vimentin,MMP-2,MMP-9,PI3K,Akt,p-Akt,mTOR,and p-mTOR was determined by Western blot,and the m RNA expression of PI3K,Akt and mTOR was determined by real-time polymerase chain reaction.RESULTS:BJ-EZ group inhibited proliferation after 24,48,and 72 h compared with the NC group(P<0.05,<0.01 or<0.001)and reduced the invasion and migration of MDA-MB-231 cells(P<0.01 or<0.001).In addition,BJ-EZ group upregulated the expression of E-cadherin,downregulated the expression of N-cadherin,Vimentin,MMP-2,and MMP-9(P<0.05,P<0.01 or P<0.001),and inhibited the m RNA and protein expression of PI3K,Akt(p-Akt),mTOR(p-mTOR)(P<0.05,<0.01 or<0.001).CONCLUSION:Biejia(Carapax Trionycis)and Ezhu(Rhizoma Curcumae Phaeocaulis)couplet medicine can inhibit the proliferation,invasion,migration and EMT of MDA-MB-231 cells through PI3K/Akt/mTOR signaling pathway,and the effect is better than that of Biejia(Carapax Trionycis)or Ezhu(Rhizoma Curcumae Phaeocaulis)alone.展开更多
目的:观察子宫内膜异位症大鼠异位内膜组织基质细胞衍生因子-1(SDF-1)及其趋化因子受体(CXCR4)的表达情况,并探讨复方莪术散对其表达的影响。方法:自体移植法建立非动情期SD大鼠子宫内膜异位症模型,PCR法检测大鼠在位、异位内膜组织及...目的:观察子宫内膜异位症大鼠异位内膜组织基质细胞衍生因子-1(SDF-1)及其趋化因子受体(CXCR4)的表达情况,并探讨复方莪术散对其表达的影响。方法:自体移植法建立非动情期SD大鼠子宫内膜异位症模型,PCR法检测大鼠在位、异位内膜组织及正常在位子宫内膜组织SDF-1和CXCR4的m RNA表达,分析各组统计学差异;造模21 d后,分组连续给药28 d,对比观察中药复方莪术散(高、中、低剂量组各10只)与模型组(10只)异位症内膜组织中SDF-1 m RNA,CXCR4 m RNA的表达差异。结果:SDF-1 m RNA和CXCR4 m RNA在大鼠异位内膜组织高表达,与在位内膜组织及正常子宫内膜组织比较有统计学差异(P<0.05);各治疗组异位内膜组织SDF-1m RNA表达均下降,高剂量组(0.646±0.264)与中、低剂量组(1.001±0.160、1.714±0.343)比较降低明显,且各组间比较有统计学意义(P<0.05);各治疗组CXCR4 m RNA表达有下降趋势,复方莪术散高剂量(0.587±0.476)和复方莪术散中剂量(1.408±0.252)组CXCR4 m RNA表达分别与模型组(3.004±0.264)比较有统计学差异(P<0.05)。结论:SDF-1、CXCR4在大鼠在位内膜组织中的表达高于正常子宫内膜组织,其表达可能与大鼠子宫内膜异位症的发病机制存在一定关系。复方莪术散可明显降低异位内膜组织中SDF-1、CXCR4表达,对子宫内膜异位症有一定治疗作用。展开更多
文摘目的:基于网络药理学及分子对接技术,探究三棱-莪术药对在辅助治疗卵巢癌中的作用机制。方法:活性成分及其作用靶点收集自中药系统药理学分析平台,卵巢癌相关靶点收集自Genecard数据库。上述靶点取交集后,结合STRING数据库及Cytoscape软件构建PPI网络。应用R及Cytoscape软件整合构建中药-活性成分-疾病-靶点网络。同时,应用R软件进行基因本体论(gene ontology,GO)功能富集、京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集,并通过Cytoscape/CluGO进行KEGG通路聚类分析。综合分析各网络,获取关键靶点及其对应活性成分后应用Autodock vina及Pymol等软件进行分子对接及可视化研究。结果:本研究在三棱-莪术药对中,共获得8个活性成分,164个作用靶点,Genecards数据库获得1540个符合条件的卵巢癌相关靶点。取交集获得27个三棱-莪术药对治疗卵巢癌的潜在靶点。分析PPI网络并获取关键靶点为CASP3、ESR1、JUN、PTGS2、AR、MAPK14、PGR、CASP8、CASP9、CAS及PPARG。GO及KEGG分析表明相关作用机制可能涉及凋亡、白细胞介素17(interleukin-17,IL-17)信号通路、乙肝信号通路、雌激素信号通路、甲状腺激素信号通路,卵巢类固醇等多条信号通路。分子对接表明核心成分芒柄花黄素、β-谷甾醇、常春藤皂苷元与关键靶点结合性良好,其中各活性成分与PTGS2结合最佳,结合能均小于-9.0 kcal/mol。结论:该研究初步揭示了三棱-莪术药对治疗卵巢癌的药理机制,这为后期探究该药对及其所在复方治疗卵巢癌提供了较为系统的依据。
基金Supported by the National Natural Science Foundation of China(No.81673810)
文摘OBJECTIVE: To study the anti-tumor effects of the extracts from Huangqi(Radix Astragali Mongolici)and Ezhu(Rhizoma Curcumae Phaeocaulis) on the growth of Lewis lung carcinoma(LLC) in a xenograft mouse model and to investigate the possible underlying mechanism.METHODS: LLC tumor-bearing C57 BL/6 mice were treated with normal saline, cisplatin(2 mg/kg intraperitoneally every other day), or Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis)(1∶1, 2∶1, or 3∶1 ratio;5, 8, or 11 g/kg crude drug intragastrically every day) for 15 d.Body weights and tumor volumes were measured every other day. Tumors were excised on day 15 and analyzed. Tumor microvessel density(MVD)was assessed by immunohistochemical staining of CD34;and expression of vascular endothelial cell growth factor(VEGF), the mitogen-activated protein kinases p38 mitogen-activated protein kinase(MAPK), extracellular signal-regulated kinases 1 and 2(ERK1/2), and Jun N-terminal kinase(JNK)and their phosphorylated forms were assessed by Western blotting.RESULTS: Treatment with cisplatin caused a significant loss of body weight compared with controls,whereas Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) extract combinations had no effect. Extracts from Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) significantly decreased tumor weight and tumor MVD compared with controls,and at the 3∶1 treatment group had similar efficacy to cisplatin in reducing MVD. Tumors from Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) treatments also showed decreased p38 MAPK, p-p38 MAPK, ERK1/2, p-ERK1/2,JNK, and p-JNK expression compared with the control group(all P < 0.01). VEGF protein expression was significantly reduced in the 2∶1 and 3∶1 treatment groups compared with the control group(P < 0.01).CONCLUSION: Extracts from Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) hindered LLC growth in the xenograft mouse model, possibly via inhibition of the MAPK signaling pathway, VEGF production, and tumor angiogenesis.
基金Supported by the National Natural Science Foundation of China:the Study on Molecular Mechanism of Anti-lung Cancer Angiogenesis of Astragalus-zedoariae in Experienced Prescriptions Based on TGF-β1/MAPK/HIF-1α Signaling Pathway(No.81673810)
文摘OBJECTIVE: To investigate the effect of optimal combination(E) of Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) on proliferation and apoptosis of A549 lung cancer cells and the possible mechanism underpinning the action.METHODS: A uniform design method was used to optimize the E of Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) in A549 lung cancer cells. MTS assay was applied to analyze the effect of the component formula ofHuangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Curcumae Phaeocaulis) on A549 cells viability in various uniform design groups. A549 cells with exponential growth in routine culture were exposed to Co Cl_2(200 μmol/L) to mimic hypoxic conditions. Group 0 was treated with RPMI-1640, the group Co Cl_2 was treated with Co Cl_2(200 μmol/L),the group DDP + Co Cl_2 was treated with 4 mg/L Cisplatin injection(DDP) + Co Cl_2(200 μmol/L), and the drug group was treated with various dose of E(0.5 E, 1 E, 2 E) + Co Cl_2(200 μmol/L). All groups were cultured for 24 h. Cell apoptosis was measured by Annexin V-FITC/propidium iodide double staining and flow cytometry. Western blot assay and quantitative real-time polymerase chain reaction(q RT-PCR) were employed to detect the protein and m RNA expression of B-celllymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax) and cysteinyl aspartate specific proteinase-3(caspase-3).RESULTS: The E obtained by the uniform design was comprise of 200 mg/L Astragalus polysaccharide(X1) and 32 mg/L Curcumin(X3). Group DDP+Co Cl_2, group 1 E + Co Cl_2 and group 2 E + Co Cl_2 promoted the apoptosis of A549 cells(P < 0.05). Group1 E + Co Cl_2 and group 2 E + Co Cl_2 had no statistically significant differences compared with the group DDP + Co Cl_2(P > 0.05). Compared with group 0, various doses of E + Co Cl_2 could up-regulate the expression of Bax and caspase-3 and down-regulate the expression of Bcl-2 at protein and m RNA levels(P < 0.05).CONCLUSION: Astragalus polysaccharide and Curcumin was the optimal combination of Huangqi(Radix Astragali Mongolici) and Ezhu(Rhizoma Cur-cumae Phaeocaulis). E promoted the apoptosis of A549 cells. Combination of Astragalus polysaccharide and Curcumin increased the expression of Bax and caspase-3, and decreased the expression of Bcl-2 to initiate apoptosis in A549 cells under chemical-induced hypoxia.
基金Supported by the National Natural Science Foundation of China:Based on the"Hu-Chang"Theory,the Mechanism of Shuyu Pill on the Effect of Epithelial-mesenchymal Transition to Inhibit the Metastasis of Triple-negative Breast Cancer by P13K/Akt/mTOR Pathway(No.81960834)。
文摘OBJECTIVE:To investigate the efficacy of Biejia(Carapax Trionycis)and Ezhu(Rhizoma Curcumae Phaeocaulis)couplet medicine on epithelial-mesenchymal transition(EMT),invasion and migration of MDA-MB-231 triple negative breast cancer(TNBC)cells based on PI3K/Akt/mTOR signaling pathway.METHODS:MDA-MB-231 cells were treated with different medicated serum as Biejia-,Ezhu-,Biejia-Ezhu(BJ-,EZ-,BJ-EZ-)groups,intervened with no drug rat serum and paclitaxel with final concentration of 33 nM(IC50)as negative and positive control(NC and PC)groups.CCK-8 assay,scratch test,and Transwell assay were used to examine cell proliferation,invasion,and migration.The expression of E-cadherin,N-cadherin,Vimentin,MMP-2,MMP-9,PI3K,Akt,p-Akt,mTOR,and p-mTOR was determined by Western blot,and the m RNA expression of PI3K,Akt and mTOR was determined by real-time polymerase chain reaction.RESULTS:BJ-EZ group inhibited proliferation after 24,48,and 72 h compared with the NC group(P<0.05,<0.01 or<0.001)and reduced the invasion and migration of MDA-MB-231 cells(P<0.01 or<0.001).In addition,BJ-EZ group upregulated the expression of E-cadherin,downregulated the expression of N-cadherin,Vimentin,MMP-2,and MMP-9(P<0.05,P<0.01 or P<0.001),and inhibited the m RNA and protein expression of PI3K,Akt(p-Akt),mTOR(p-mTOR)(P<0.05,<0.01 or<0.001).CONCLUSION:Biejia(Carapax Trionycis)and Ezhu(Rhizoma Curcumae Phaeocaulis)couplet medicine can inhibit the proliferation,invasion,migration and EMT of MDA-MB-231 cells through PI3K/Akt/mTOR signaling pathway,and the effect is better than that of Biejia(Carapax Trionycis)or Ezhu(Rhizoma Curcumae Phaeocaulis)alone.
文摘目的:观察子宫内膜异位症大鼠异位内膜组织基质细胞衍生因子-1(SDF-1)及其趋化因子受体(CXCR4)的表达情况,并探讨复方莪术散对其表达的影响。方法:自体移植法建立非动情期SD大鼠子宫内膜异位症模型,PCR法检测大鼠在位、异位内膜组织及正常在位子宫内膜组织SDF-1和CXCR4的m RNA表达,分析各组统计学差异;造模21 d后,分组连续给药28 d,对比观察中药复方莪术散(高、中、低剂量组各10只)与模型组(10只)异位症内膜组织中SDF-1 m RNA,CXCR4 m RNA的表达差异。结果:SDF-1 m RNA和CXCR4 m RNA在大鼠异位内膜组织高表达,与在位内膜组织及正常子宫内膜组织比较有统计学差异(P<0.05);各治疗组异位内膜组织SDF-1m RNA表达均下降,高剂量组(0.646±0.264)与中、低剂量组(1.001±0.160、1.714±0.343)比较降低明显,且各组间比较有统计学意义(P<0.05);各治疗组CXCR4 m RNA表达有下降趋势,复方莪术散高剂量(0.587±0.476)和复方莪术散中剂量(1.408±0.252)组CXCR4 m RNA表达分别与模型组(3.004±0.264)比较有统计学差异(P<0.05)。结论:SDF-1、CXCR4在大鼠在位内膜组织中的表达高于正常子宫内膜组织,其表达可能与大鼠子宫内膜异位症的发病机制存在一定关系。复方莪术散可明显降低异位内膜组织中SDF-1、CXCR4表达,对子宫内膜异位症有一定治疗作用。
