【目的】探讨桃核承气汤对重症急性胰腺炎(SAP)大鼠肠道功能的影响及作用机制。【方法】将50只SD大鼠随机分为空白对照组、SAP模型组、桃核承气汤组(灌胃剂量为23.5 m L·kg-1·d-1)、奥曲肽组(腹腔注射剂量为10μg·kg-1&#...【目的】探讨桃核承气汤对重症急性胰腺炎(SAP)大鼠肠道功能的影响及作用机制。【方法】将50只SD大鼠随机分为空白对照组、SAP模型组、桃核承气汤组(灌胃剂量为23.5 m L·kg-1·d-1)、奥曲肽组(腹腔注射剂量为10μg·kg-1·d-1)、联合用药组(桃核承气汤灌胃剂量为23.5 m L·kg-1·d-1,奥曲肽腹腔注射剂量为10μg·kg-1·d-1)。除空白对照组外,其余各组采用L-精氨酸腹腔注射复制大鼠SAP模型,造模成功后24 h取材,采用菌群记数法检测各组大鼠粪便菌群变化,生化测定各组大鼠血清淀粉酶(AMY)水平,采用酶联免疫吸附(ELISA)法检测各组大鼠血清白细胞介素(IL)-6、IL-10、肿瘤坏死因子(TNF)-α水平,采用苏木素—伊红(HE)染色法观察胰腺和大肠组织病理变化。【结果】与空白对照组比较,SAP模型组大鼠血清AMY、TNF-α、IL-6水平显著降低,IL-10水平显著升高(P<0.05),肠球菌、肠杆菌增多,乳酸杆菌、双歧杆菌则大幅度骤减,胰腺、大肠组织结构破坏,大量炎症细胞浸润。与SAP模型组比较,3个治疗组大鼠血清AMY、TNF-α、IL-6水平显著升高(P<0.05),IL-10水平显著降低(P<0.05),肠球菌、肠杆菌减少,乳酸杆菌、双歧杆菌增加,胰腺、大肠组织病理变化均减轻,以联合用药组最明显。【结论】桃核承气汤对SAP大鼠的肠道功能有良好的改善作用,可减轻炎症反应,改善肠道菌群,抑制SAP病情的恶化。展开更多
Objective： To observe the anti-virus effects of andrographolide （AD） on the retinoic acid-inducible gene-I （RIG-I）-Iike receptors （RLRs） signaling pathway when immunological cells were infected with HIN1. Meth...Objective： To observe the anti-virus effects of andrographolide （AD） on the retinoic acid-inducible gene-I （RIG-I）-Iike receptors （RLRs） signaling pathway when immunological cells were infected with HIN1. Methods： Leukomonocyte was obtained from umbilical cord blood by Ficoll density gradient centrifugation, and immunological cells were harvested after cytokines stimulation. Virus infected cell model was established by H1N1 co-cultured with normal human bronchial epithelial cell line （16HBE）. The optimal concentration of AD was defined by methyl-thiazolyl-tetrazolium （MTT） assay. After the virus infected cell model was established, AD was added into the medium as a treatment intervention. After 24-h co-culture, cell supernatant was collected for interferon gamma （IFN- ~, ） and interleukin-4 （IL-4） enzyme-linked immunosorbent assay （ELISA） detection while immunological cells for real-time polymerase chain reaction （RT-PCR）. Results： The optimal concentration of AD for anti-virus effect was 250 μg/mL. IL-4 and IFN-γ in the supernatant and mRNA levels in RLRs pathway increased when cells was infected by virus, RIG-I, IFN-13 promoter stimulator-1 （IPS-1）, interferon regulatory factor （IRF）-7, IRF-3 and nuclear transcription factor K B （NF- K B） mRNA levels increased significantly （P〈0.05）. When AD was added into co-culture medium, the levels of IL-4 and IFN-γ were lower than those in the non-interference groups and the mRNA expression levels decreased, RIG-I, IPS-1, IRF-7, IRF-3 and NF- K B decreased significantly in each group with significant statistic differences （P〈0.05）. Conclusions： The RLRs mediated viral recognition provided a potential molecular target for acute viral infections and andrographolide could ameliorate H1N1 virus-induced cell mortality. And the antiviral effects might be related to its inhibition of viral-induced activation of the RLRs signaling pathway.展开更多
Objective：To study the immunologic function of dendritic cells（DCs） cultured in two kinds of hepatoma cell line＇s supernatant and the enhancing effects of carboxymethylpachymaran（CMP） on DCs.Methods：DCs were ha...Objective：To study the immunologic function of dendritic cells（DCs） cultured in two kinds of hepatoma cell line＇s supernatant and the enhancing effects of carboxymethylpachymaran（CMP） on DCs.Methods：DCs were harvested after stimulation by granulocyte-macrophage colony-stimulating factor （GM-CSF） and interleukin（IL）-4 from umbilical cord blood using density-gradient centrifugation method. Cultured supernatant of two hepatoma cell lines（HepG2 and HepG2.2.15） were collected for condition medium（CM） according to a volume ratio of supernatant to incomplete RPMI-1640 medium,which was 3：1.CMP was dissolved in incomplete RPMI-1640 medium.Experimental groups were divided according to the culture medium,either CM or with CMP in it.DCs subsets CD83,CD86,CD1a,and d-related human leukocyte antigens（HLA-DR） were analyzed by flow cytometry.The proliferation ability of allogeneic T cells in mixed lymphocyte reaction（MLR） stimulated by DCs was examined by 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide（MTT） analysis.IL-12p70,interferon-γ（IFN-γ）,and nuclear factor k B（NF- k B） were detected by enzyme-linked immunosorbent assay analysis.Results：The proliferation of lymphocytes and secreting level of IL-12 and expression of phenotype of DCs cultured in two kinds of CM were lower than those of normal group（P0.01）.Compared with the normal group,groups treated with CMP showed a higher expression level of DCs subsets,lymphocyte reproductive activity,as well as IL-12 and IFN-γsecretion levels.Groups treated with CMP also demonstrated higher levels of DCs phenotype expression and IL-12 and IFN-γsecretion in supernatant of MLR and higher lymphocyte reproductive activity compared with CM group（P0.05）.Compared with the normal group,the expression level of NF-k B in DCs nuclear was higher in CMP groups but lower in two CM groups（P0.05）.After CMP was added,the NF-κB expression levels of two CM groups were increased compared with levels before CMP was added展开更多
文摘【目的】探讨桃核承气汤对重症急性胰腺炎(SAP)大鼠肠道功能的影响及作用机制。【方法】将50只SD大鼠随机分为空白对照组、SAP模型组、桃核承气汤组(灌胃剂量为23.5 m L·kg-1·d-1)、奥曲肽组(腹腔注射剂量为10μg·kg-1·d-1)、联合用药组(桃核承气汤灌胃剂量为23.5 m L·kg-1·d-1,奥曲肽腹腔注射剂量为10μg·kg-1·d-1)。除空白对照组外,其余各组采用L-精氨酸腹腔注射复制大鼠SAP模型,造模成功后24 h取材,采用菌群记数法检测各组大鼠粪便菌群变化,生化测定各组大鼠血清淀粉酶(AMY)水平,采用酶联免疫吸附(ELISA)法检测各组大鼠血清白细胞介素(IL)-6、IL-10、肿瘤坏死因子(TNF)-α水平,采用苏木素—伊红(HE)染色法观察胰腺和大肠组织病理变化。【结果】与空白对照组比较,SAP模型组大鼠血清AMY、TNF-α、IL-6水平显著降低,IL-10水平显著升高(P<0.05),肠球菌、肠杆菌增多,乳酸杆菌、双歧杆菌则大幅度骤减,胰腺、大肠组织结构破坏,大量炎症细胞浸润。与SAP模型组比较,3个治疗组大鼠血清AMY、TNF-α、IL-6水平显著升高(P<0.05),IL-10水平显著降低(P<0.05),肠球菌、肠杆菌减少,乳酸杆菌、双歧杆菌增加,胰腺、大肠组织病理变化均减轻,以联合用药组最明显。【结论】桃核承气汤对SAP大鼠的肠道功能有良好的改善作用,可减轻炎症反应,改善肠道菌群,抑制SAP病情的恶化。
基金Supported by the National Natural Science Foundation of China(No.81273616 and 30973693)the Doctoral Fund of the Ministryof Education 2010(No.20104401110003)the Natural Science Foundation of Guangdong Province(No.S2013010013434)
文摘Objective： To observe the anti-virus effects of andrographolide （AD） on the retinoic acid-inducible gene-I （RIG-I）-Iike receptors （RLRs） signaling pathway when immunological cells were infected with HIN1. Methods： Leukomonocyte was obtained from umbilical cord blood by Ficoll density gradient centrifugation, and immunological cells were harvested after cytokines stimulation. Virus infected cell model was established by H1N1 co-cultured with normal human bronchial epithelial cell line （16HBE）. The optimal concentration of AD was defined by methyl-thiazolyl-tetrazolium （MTT） assay. After the virus infected cell model was established, AD was added into the medium as a treatment intervention. After 24-h co-culture, cell supernatant was collected for interferon gamma （IFN- ~, ） and interleukin-4 （IL-4） enzyme-linked immunosorbent assay （ELISA） detection while immunological cells for real-time polymerase chain reaction （RT-PCR）. Results： The optimal concentration of AD for anti-virus effect was 250 μg/mL. IL-4 and IFN-γ in the supernatant and mRNA levels in RLRs pathway increased when cells was infected by virus, RIG-I, IFN-13 promoter stimulator-1 （IPS-1）, interferon regulatory factor （IRF）-7, IRF-3 and nuclear transcription factor K B （NF- K B） mRNA levels increased significantly （P〈0.05）. When AD was added into co-culture medium, the levels of IL-4 and IFN-γ were lower than those in the non-interference groups and the mRNA expression levels decreased, RIG-I, IPS-1, IRF-7, IRF-3 and NF- K B decreased significantly in each group with significant statistic differences （P〈0.05）. Conclusions： The RLRs mediated viral recognition provided a potential molecular target for acute viral infections and andrographolide could ameliorate H1N1 virus-induced cell mortality. And the antiviral effects might be related to its inhibition of viral-induced activation of the RLRs signaling pathway.
基金Supported by the National Natural Science Foundation of China (No.30973693)the Research Fund for the Doctoral Program of Higher Education of China(No.20104401110003)
文摘Objective：To study the immunologic function of dendritic cells（DCs） cultured in two kinds of hepatoma cell line＇s supernatant and the enhancing effects of carboxymethylpachymaran（CMP） on DCs.Methods：DCs were harvested after stimulation by granulocyte-macrophage colony-stimulating factor （GM-CSF） and interleukin（IL）-4 from umbilical cord blood using density-gradient centrifugation method. Cultured supernatant of two hepatoma cell lines（HepG2 and HepG2.2.15） were collected for condition medium（CM） according to a volume ratio of supernatant to incomplete RPMI-1640 medium,which was 3：1.CMP was dissolved in incomplete RPMI-1640 medium.Experimental groups were divided according to the culture medium,either CM or with CMP in it.DCs subsets CD83,CD86,CD1a,and d-related human leukocyte antigens（HLA-DR） were analyzed by flow cytometry.The proliferation ability of allogeneic T cells in mixed lymphocyte reaction（MLR） stimulated by DCs was examined by 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide（MTT） analysis.IL-12p70,interferon-γ（IFN-γ）,and nuclear factor k B（NF- k B） were detected by enzyme-linked immunosorbent assay analysis.Results：The proliferation of lymphocytes and secreting level of IL-12 and expression of phenotype of DCs cultured in two kinds of CM were lower than those of normal group（P0.01）.Compared with the normal group,groups treated with CMP showed a higher expression level of DCs subsets,lymphocyte reproductive activity,as well as IL-12 and IFN-γsecretion levels.Groups treated with CMP also demonstrated higher levels of DCs phenotype expression and IL-12 and IFN-γsecretion in supernatant of MLR and higher lymphocyte reproductive activity compared with CM group（P0.05）.Compared with the normal group,the expression level of NF-k B in DCs nuclear was higher in CMP groups but lower in two CM groups（P0.05）.After CMP was added,the NF-κB expression levels of two CM groups were increased compared with levels before CMP was added
