Objective: To study the mechanism of Sijunzi decoction treating limb weakness in spleen Qi deficiency (SQD) based on the myonuclear domain (MND) theory. Methods: 40 male Sprague-Dawley rats were randomly divided into ...Objective: To study the mechanism of Sijunzi decoction treating limb weakness in spleen Qi deficiency (SQD) based on the myonuclear domain (MND) theory. Methods: 40 male Sprague-Dawley rats were randomly divided into the normal group, SQD model group (model group), SQD+ still water group (SW group) and SQD+ Sijunzi decoction group (CM group), 10 rats each group;Grip-Strength Meter was used to measure limb grip strength;transmission electron microscope was employed to observe the ultrastructural changes of the myofibers, Image Pro 6.0 was used to measure the myonuclear numbers, cross-section area (CSA) and then their ratios (the MND sizes) were calculated, immunofluorescence assay was chosen to test the expressions of paired box gene 7 (Pax7) and myogenic differentiation antigen (MyoD). Results: Compared with those in the normal group, limb grip strength was decreased, sarcomeres were abnormal, and all the myonuclear numbers, CSA and MND sizes were reduced, but the Pax7+ cell numbers were increased, significantly, in the model and SW groups;Compared with those in the model and SW groups, limb grip strength was increased, sarcomeres were basically normal, the myonuclear number and CSA were both greater, and the Pax7+ and MyoD+ cell numbers were both increased, significantly, in the CM group. Conclusion: Sijunzi decoction might increase the myonuclear number by activating the MSCs to treat limb weakness in SQD.展开更多
Sijunzi Decoction,as one of the classic Chinese traditional prescriptions,has been used clinically by major physicians since the Song Dynasty.This article reviewed and sorted out the literature on the effective chemis...Sijunzi Decoction,as one of the classic Chinese traditional prescriptions,has been used clinically by major physicians since the Song Dynasty.This article reviewed and sorted out the literature on the effective chemistry of Sijunzi Decoction and the mechanism of its prevention and treatment of digestive system diseases.At present,its effective chemical components are derived from the saponins and flavonoids in ginseng and licorice,and are effective for gastrointestinal mucosal injury diseases and malignant digestive system.Digestive system diseases such as tumors,functional gastrointestinal diseases,non-alcoholic fatty liver,acute liver injury,and liver failure show a multi-path,multi-target effect mechanism.This article reviews the effective chemical components and research of Sijunzi Decoction and the related mechanisms of prevention and treatment of digestive system diseases,and provides valuable clues for the follow-up research of Sijunzi Decoction.展开更多
Aim An HPLC-UV method for the analysis of Traditional Chinese Medicine （TCM） preparation, Sijunzi decoction, has been developed. Four flavonoid marker compounds, liquiritigenin, liquiritin, isoliquiritigenin, and is...Aim An HPLC-UV method for the analysis of Traditional Chinese Medicine （TCM） preparation, Sijunzi decoction, has been developed. Four flavonoid marker compounds, liquiritigenin, liquiritin, isoliquiritigenin, and isoliquiritin, from Radix Glycyrrhizae were quantitatively analyzed in this preparation. Methods The separation was performed on a reversed-phase C18 column by using a gradient elution with mobile phase of （A） water-formic acid （100∶0.04, V/V） （pH 3） and （B） acetonitrile. The mobile phase gradient was set from 0-5 min at 10% of B and 45 min to 90% of B. The assay was carried out at a flow rate of 0.2 mL·min-1 at room temperature with the UV detection wavelengths at 280 nm and 368 nm. Results The extract （25 mg·mL^-1） of Sijunzi decoction contains 1.47 μg·mL^-1 liquiritigenin, 16.40 μg·mL^-1 liquiritin, 0.66 μg·mL^-1 isoliquiritigenin, and 2.12 μg·mL^-1 isoliquiritin. The recoveries for the sample preparation of the markers were 102.2%, 97.7%, 100.3%, and 99.9%, respectively. Conclusion The method is simple and accurate. This study reports a routine quantitative method for the analysis of multiple components in Sijunzi decoction by HPLC.展开更多
目的:运用网络药理学分析四君子汤辅助治疗胃癌的作用机制,并进行分子对接验证。方法:通过中药系统药理学数据库和分析平台提取四君子汤中各药物化学成分及相关靶点,并利用常用疾病数据库筛选出胃癌靶点,绘制药物-疾病共同靶点的韦恩图...目的:运用网络药理学分析四君子汤辅助治疗胃癌的作用机制,并进行分子对接验证。方法:通过中药系统药理学数据库和分析平台提取四君子汤中各药物化学成分及相关靶点,并利用常用疾病数据库筛选出胃癌靶点,绘制药物-疾病共同靶点的韦恩图。再用Cytoscape软件构建药物-疾病调控网络,在STRING数据库中构建药物-疾病共同靶点的蛋白质相互作用网络(protein-protein interaction,PPI),在Metascape数据库进行基因本体(gene ontology,GO)生物过程富集分析和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。通过分子对接验证前期所得的关键药物化学成分与核心靶点的结合可能性。结果:得到四君子汤135个药物化学成分,胃癌作用靶点1423个,药物-疾病共同作用靶点65个;通过药物-疾病调控网络和拓扑分析,发现药物化学成分中度值较高的为槲皮素、山柰酚、异鼠李素等;PPI网络核心靶点为MAPK8、ESR1、CCND1等;GO富集分析显示,四君子汤主要通过影响细胞RNA聚合酶Ⅱ启动子转录的正调控、凋亡过程的负调控、DNA模板的转录正调控等发挥生物学和药理学的多种效应;相应的KEGG通路分析显著富集于癌症途径、p53信号通路、NF-κB信号通路等多条与胃癌密切相关的通路;分子对接验证结合性好。结论:网络药理学初步明确了四君子汤辅助治疗胃癌潜在的靶点通路机制,分子对接角度进一步验证了研究思路的正确性。展开更多
目的探讨四君子汤对抗PD-1免疫治疗小鼠三阴性乳腺癌(triple negative breast cancer,TNBC)的疗效及作用机制。方法建立TNBC移植瘤小鼠模型,将小鼠随机分为4T1组(模型组)、四君子汤组、抗PD-1治疗组、四君子汤联合抗PD-1治疗组,每组6只...目的探讨四君子汤对抗PD-1免疫治疗小鼠三阴性乳腺癌(triple negative breast cancer,TNBC)的疗效及作用机制。方法建立TNBC移植瘤小鼠模型,将小鼠随机分为4T1组(模型组)、四君子汤组、抗PD-1治疗组、四君子汤联合抗PD-1治疗组,每组6只。四君子汤组小鼠按实验动物与人体表面积比换算给予4.85 g/(kg·d)四君子汤灌胃(每周5次),抗PD-1治疗组给予80μg/只抗PD-1抗体腹腔注射(每周2次),四君子汤联合抗PD-1治疗组则分别给予上述治疗,4T1组则灌胃等体积的0.9%氯化钠溶液(每周5次);连续干预3周。比较各组肿瘤体积大小,通过生存分析比较各组小鼠生存时间。采用C11-BODIPY染色法进行脂质过氧化评估各组小鼠肿瘤细胞脂质过氧化水平,并通过测量丙二醛(malondialdehyde,MDA)含量确认;采用7-氨基放线菌素D(7-aminoactinomycin D,7-AAD)染色法检测细胞死亡比例;采用实时定量PCR检测铁死亡相关基因表达。并进一步通过体外实验分析四君子汤有效成分(人参皂苷Rb1)对4T1癌细胞脂质过氧化和铁死亡的影响。结果与抗PD-1治疗组相比,四君子汤联合抗PD-1治疗组小鼠肿瘤体积明显缩小(P<0.01),四君子汤联合抗PD-1治疗组中位生存期明显延长(34 d vs.25 d,P<0.05)。与抗PD-1治疗组相比,四君子汤联合抗PD-1治疗组脂质过氧化平均荧光强度明显升高(P<0.05),肿瘤细胞中的MDA含量、癌细胞死亡率均明显升高(P均<0.01)。四君子汤组(与4T1组相比)和四君子汤联合抗PD-1治疗组(与抗PD-1治疗组相比)铁死亡抑制基因NRF2、GPX4、SLC7A11表达水平显著降低(P均<0.05),但铁死亡促进基因TFRC组间比较差异无统计学意义(P>0.05)。细胞实验结果显示:与4T1细胞空白对照组相比,人参皂苷Rb1单体对乳腺癌细胞脂质过氧化平均荧光强度的作用差异无统计学意义(P>0.05)。人参皂苷Rb1单体联合IFN-γ对乳腺癌细胞脂质过氧化平均荧光强度的作用明显高于单用IFN-γ(P<0.05)。人参皂苷Rb1干预下4T1癌细胞铁死亡抑制基因NRF2、GPX4、SLC7A11表达水平显著降低(P均<0.05)。结论四君子汤单独应用未见对小鼠TNBC有明显治疗效果,但可增强抗PD-1抗体免疫治疗小鼠TNBC的效果,其机制可能与四君子汤降低癌细胞铁死亡抑制基因表达有关。展开更多
文摘Objective: To study the mechanism of Sijunzi decoction treating limb weakness in spleen Qi deficiency (SQD) based on the myonuclear domain (MND) theory. Methods: 40 male Sprague-Dawley rats were randomly divided into the normal group, SQD model group (model group), SQD+ still water group (SW group) and SQD+ Sijunzi decoction group (CM group), 10 rats each group;Grip-Strength Meter was used to measure limb grip strength;transmission electron microscope was employed to observe the ultrastructural changes of the myofibers, Image Pro 6.0 was used to measure the myonuclear numbers, cross-section area (CSA) and then their ratios (the MND sizes) were calculated, immunofluorescence assay was chosen to test the expressions of paired box gene 7 (Pax7) and myogenic differentiation antigen (MyoD). Results: Compared with those in the normal group, limb grip strength was decreased, sarcomeres were abnormal, and all the myonuclear numbers, CSA and MND sizes were reduced, but the Pax7+ cell numbers were increased, significantly, in the model and SW groups;Compared with those in the model and SW groups, limb grip strength was increased, sarcomeres were basically normal, the myonuclear number and CSA were both greater, and the Pax7+ and MyoD+ cell numbers were both increased, significantly, in the CM group. Conclusion: Sijunzi decoction might increase the myonuclear number by activating the MSCs to treat limb weakness in SQD.
文摘Sijunzi Decoction,as one of the classic Chinese traditional prescriptions,has been used clinically by major physicians since the Song Dynasty.This article reviewed and sorted out the literature on the effective chemistry of Sijunzi Decoction and the mechanism of its prevention and treatment of digestive system diseases.At present,its effective chemical components are derived from the saponins and flavonoids in ginseng and licorice,and are effective for gastrointestinal mucosal injury diseases and malignant digestive system.Digestive system diseases such as tumors,functional gastrointestinal diseases,non-alcoholic fatty liver,acute liver injury,and liver failure show a multi-path,multi-target effect mechanism.This article reviews the effective chemical components and research of Sijunzi Decoction and the related mechanisms of prevention and treatment of digestive system diseases,and provides valuable clues for the follow-up research of Sijunzi Decoction.
文摘Aim An HPLC-UV method for the analysis of Traditional Chinese Medicine （TCM） preparation, Sijunzi decoction, has been developed. Four flavonoid marker compounds, liquiritigenin, liquiritin, isoliquiritigenin, and isoliquiritin, from Radix Glycyrrhizae were quantitatively analyzed in this preparation. Methods The separation was performed on a reversed-phase C18 column by using a gradient elution with mobile phase of （A） water-formic acid （100∶0.04, V/V） （pH 3） and （B） acetonitrile. The mobile phase gradient was set from 0-5 min at 10% of B and 45 min to 90% of B. The assay was carried out at a flow rate of 0.2 mL·min-1 at room temperature with the UV detection wavelengths at 280 nm and 368 nm. Results The extract （25 mg·mL^-1） of Sijunzi decoction contains 1.47 μg·mL^-1 liquiritigenin, 16.40 μg·mL^-1 liquiritin, 0.66 μg·mL^-1 isoliquiritigenin, and 2.12 μg·mL^-1 isoliquiritin. The recoveries for the sample preparation of the markers were 102.2%, 97.7%, 100.3%, and 99.9%, respectively. Conclusion The method is simple and accurate. This study reports a routine quantitative method for the analysis of multiple components in Sijunzi decoction by HPLC.
文摘目的:运用网络药理学分析四君子汤辅助治疗胃癌的作用机制,并进行分子对接验证。方法:通过中药系统药理学数据库和分析平台提取四君子汤中各药物化学成分及相关靶点,并利用常用疾病数据库筛选出胃癌靶点,绘制药物-疾病共同靶点的韦恩图。再用Cytoscape软件构建药物-疾病调控网络,在STRING数据库中构建药物-疾病共同靶点的蛋白质相互作用网络(protein-protein interaction,PPI),在Metascape数据库进行基因本体(gene ontology,GO)生物过程富集分析和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。通过分子对接验证前期所得的关键药物化学成分与核心靶点的结合可能性。结果:得到四君子汤135个药物化学成分,胃癌作用靶点1423个,药物-疾病共同作用靶点65个;通过药物-疾病调控网络和拓扑分析,发现药物化学成分中度值较高的为槲皮素、山柰酚、异鼠李素等;PPI网络核心靶点为MAPK8、ESR1、CCND1等;GO富集分析显示,四君子汤主要通过影响细胞RNA聚合酶Ⅱ启动子转录的正调控、凋亡过程的负调控、DNA模板的转录正调控等发挥生物学和药理学的多种效应;相应的KEGG通路分析显著富集于癌症途径、p53信号通路、NF-κB信号通路等多条与胃癌密切相关的通路;分子对接验证结合性好。结论:网络药理学初步明确了四君子汤辅助治疗胃癌潜在的靶点通路机制,分子对接角度进一步验证了研究思路的正确性。
文摘目的探讨四君子汤对抗PD-1免疫治疗小鼠三阴性乳腺癌(triple negative breast cancer,TNBC)的疗效及作用机制。方法建立TNBC移植瘤小鼠模型,将小鼠随机分为4T1组(模型组)、四君子汤组、抗PD-1治疗组、四君子汤联合抗PD-1治疗组,每组6只。四君子汤组小鼠按实验动物与人体表面积比换算给予4.85 g/(kg·d)四君子汤灌胃(每周5次),抗PD-1治疗组给予80μg/只抗PD-1抗体腹腔注射(每周2次),四君子汤联合抗PD-1治疗组则分别给予上述治疗,4T1组则灌胃等体积的0.9%氯化钠溶液(每周5次);连续干预3周。比较各组肿瘤体积大小,通过生存分析比较各组小鼠生存时间。采用C11-BODIPY染色法进行脂质过氧化评估各组小鼠肿瘤细胞脂质过氧化水平,并通过测量丙二醛(malondialdehyde,MDA)含量确认;采用7-氨基放线菌素D(7-aminoactinomycin D,7-AAD)染色法检测细胞死亡比例;采用实时定量PCR检测铁死亡相关基因表达。并进一步通过体外实验分析四君子汤有效成分(人参皂苷Rb1)对4T1癌细胞脂质过氧化和铁死亡的影响。结果与抗PD-1治疗组相比,四君子汤联合抗PD-1治疗组小鼠肿瘤体积明显缩小(P<0.01),四君子汤联合抗PD-1治疗组中位生存期明显延长(34 d vs.25 d,P<0.05)。与抗PD-1治疗组相比,四君子汤联合抗PD-1治疗组脂质过氧化平均荧光强度明显升高(P<0.05),肿瘤细胞中的MDA含量、癌细胞死亡率均明显升高(P均<0.01)。四君子汤组(与4T1组相比)和四君子汤联合抗PD-1治疗组(与抗PD-1治疗组相比)铁死亡抑制基因NRF2、GPX4、SLC7A11表达水平显著降低(P均<0.05),但铁死亡促进基因TFRC组间比较差异无统计学意义(P>0.05)。细胞实验结果显示:与4T1细胞空白对照组相比,人参皂苷Rb1单体对乳腺癌细胞脂质过氧化平均荧光强度的作用差异无统计学意义(P>0.05)。人参皂苷Rb1单体联合IFN-γ对乳腺癌细胞脂质过氧化平均荧光强度的作用明显高于单用IFN-γ(P<0.05)。人参皂苷Rb1干预下4T1癌细胞铁死亡抑制基因NRF2、GPX4、SLC7A11表达水平显著降低(P均<0.05)。结论四君子汤单独应用未见对小鼠TNBC有明显治疗效果,但可增强抗PD-1抗体免疫治疗小鼠TNBC的效果,其机制可能与四君子汤降低癌细胞铁死亡抑制基因表达有关。
