Background:Diabetic kidney disease(DKD)is a microvascular complication of diabetes mellitus and is the main cause of end-stage renal failure.Suoquan pills(SQP)has a variety of pharmacological activities and multiple t...Background:Diabetic kidney disease(DKD)is a microvascular complication of diabetes mellitus and is the main cause of end-stage renal failure.Suoquan pills(SQP)has a variety of pharmacological activities and multiple therapeutic effects,and it is used clinically as a basic formula for the treatment of DKD.Methods:Public databases were used to identify SQP compounds and the potential targets of SQP and DKD.A drug-component-therapeutic target network was constructed.Protein-protein interaction network analysis,Gene Ontology functional analysis,and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were used to analyse the potential molecular mechanisms of SQP based on common targets of drugs and diseases.Molecular docking simulations were conducted to confirm the binding abity of the core compounds to key targets.The efficacy and predicted molecular mechanisms of SQP were validated using cell counting kit-8 assay,flow cytometry,and western blotting with HK-2 cells as a model.Results:Network pharmacology analysis showed that 26 compounds and 207 potential targets of SQP were involved in the treatment of DKD;boldine,denudatin B,pinocembrin,kaempferoid,and quercetin were considered core compounds,and epidermal growth factor receptor(EGFR)and proto-oncogene,non-receptor tyrosine kinase(SRC)were considered key targets.Gene Ontology enrichment analysis indicated that protein phosphorylation and negative regulation of apoptotic processes are important biological processes in the treatment of DKD by SQP.Molecular docking confirmed the excellent binding abilities of boldine,denudatin B,kaempferide,and quercetin to EGFR and SRC.The results of in vitro experiments showed that treatment with an ethanolic extract of SQP significantly protected HK-2 cells from high glucose-induced cell damage.In addition,the SQP ethanol extract inhibited the phosphorylation of EGFR and SRC,suppressed the apoptosis rate,and regulated apoptosis-related proteins in HK-2 cells under high glucose stress.Conclusion:This study systematically and intuitively illustrated the possible pharmacological mechanisms of SQP against DKD through multiple components,targets,and signalling pathways,especially the inhibition of EGFR and SRC phosphorylation and apoptosis.展开更多
【目的】观察自然衰老大鼠膀胱逼尿肌中腺苷酸环化酶(AC)、环磷酸腺苷(cAMP)及蛋白激酶A(PKA)的浓度变化及PKA蛋白表达,探讨缩泉丸"缩尿"的作用机理。【方法】选用老年大鼠(20月龄)50只,分为模型组,金匮肾气丸组,缩泉丸高、...【目的】观察自然衰老大鼠膀胱逼尿肌中腺苷酸环化酶(AC)、环磷酸腺苷(cAMP)及蛋白激酶A(PKA)的浓度变化及PKA蛋白表达,探讨缩泉丸"缩尿"的作用机理。【方法】选用老年大鼠(20月龄)50只,分为模型组,金匮肾气丸组,缩泉丸高、中、低剂量组,每组10只;青年大鼠(3月龄)10只作为空白组。采用酶联免疫吸附(ELISA)法检测逼尿肌中AC活性、cAMP及PKA的浓度变化,采用W estern b lotting检测逼尿肌PKA蛋白的表达。【结果】与模型组比较,缩泉丸组可显著提高自然衰老大鼠逼尿肌中AC活性、cAMP及PKA含量并上调PKA蛋白的表达。【结论】缩泉丸治疗肾虚多尿的作用与其能增加自然衰老大鼠逼尿肌中AC活性、cAMP及PKA含量并增加PKA蛋白的表达有关。展开更多
目的研究不同厂家和不同批次缩泉丸的HPLC指纹图谱,为完善该制剂的质量评价体系奠定基础。方法采用甲醇超声提取的方法制备供试品溶液,以乙腈-0.1%磷酸溶液梯度洗脱,流速:1 m L·min-1,柱温30℃,分段切换检测波长。结果建立了缩泉丸...目的研究不同厂家和不同批次缩泉丸的HPLC指纹图谱,为完善该制剂的质量评价体系奠定基础。方法采用甲醇超声提取的方法制备供试品溶液,以乙腈-0.1%磷酸溶液梯度洗脱,流速:1 m L·min-1,柱温30℃,分段切换检测波长。结果建立了缩泉丸的HPLC指纹图谱共有模式,确立了14个共有峰,15批样品指纹图谱中各共有峰相对保留时间的相对标准偏差均<1.5%。结论该方法准确可靠、重复性好,建立的指纹图谱为缩泉丸质量评价提供依据。展开更多
基金supported by the grants from National Natural Science Foundation of China(No.82174334)Hainan Province in 2022 postgraduate innovation research projects(No.Qhys2022-273).
文摘Background:Diabetic kidney disease(DKD)is a microvascular complication of diabetes mellitus and is the main cause of end-stage renal failure.Suoquan pills(SQP)has a variety of pharmacological activities and multiple therapeutic effects,and it is used clinically as a basic formula for the treatment of DKD.Methods:Public databases were used to identify SQP compounds and the potential targets of SQP and DKD.A drug-component-therapeutic target network was constructed.Protein-protein interaction network analysis,Gene Ontology functional analysis,and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were used to analyse the potential molecular mechanisms of SQP based on common targets of drugs and diseases.Molecular docking simulations were conducted to confirm the binding abity of the core compounds to key targets.The efficacy and predicted molecular mechanisms of SQP were validated using cell counting kit-8 assay,flow cytometry,and western blotting with HK-2 cells as a model.Results:Network pharmacology analysis showed that 26 compounds and 207 potential targets of SQP were involved in the treatment of DKD;boldine,denudatin B,pinocembrin,kaempferoid,and quercetin were considered core compounds,and epidermal growth factor receptor(EGFR)and proto-oncogene,non-receptor tyrosine kinase(SRC)were considered key targets.Gene Ontology enrichment analysis indicated that protein phosphorylation and negative regulation of apoptotic processes are important biological processes in the treatment of DKD by SQP.Molecular docking confirmed the excellent binding abilities of boldine,denudatin B,kaempferide,and quercetin to EGFR and SRC.The results of in vitro experiments showed that treatment with an ethanolic extract of SQP significantly protected HK-2 cells from high glucose-induced cell damage.In addition,the SQP ethanol extract inhibited the phosphorylation of EGFR and SRC,suppressed the apoptosis rate,and regulated apoptosis-related proteins in HK-2 cells under high glucose stress.Conclusion:This study systematically and intuitively illustrated the possible pharmacological mechanisms of SQP against DKD through multiple components,targets,and signalling pathways,especially the inhibition of EGFR and SRC phosphorylation and apoptosis.
文摘【目的】观察自然衰老大鼠膀胱逼尿肌中腺苷酸环化酶(AC)、环磷酸腺苷(cAMP)及蛋白激酶A(PKA)的浓度变化及PKA蛋白表达,探讨缩泉丸"缩尿"的作用机理。【方法】选用老年大鼠(20月龄)50只,分为模型组,金匮肾气丸组,缩泉丸高、中、低剂量组,每组10只;青年大鼠(3月龄)10只作为空白组。采用酶联免疫吸附(ELISA)法检测逼尿肌中AC活性、cAMP及PKA的浓度变化,采用W estern b lotting检测逼尿肌PKA蛋白的表达。【结果】与模型组比较,缩泉丸组可显著提高自然衰老大鼠逼尿肌中AC活性、cAMP及PKA含量并上调PKA蛋白的表达。【结论】缩泉丸治疗肾虚多尿的作用与其能增加自然衰老大鼠逼尿肌中AC活性、cAMP及PKA含量并增加PKA蛋白的表达有关。
文摘目的研究不同厂家和不同批次缩泉丸的HPLC指纹图谱,为完善该制剂的质量评价体系奠定基础。方法采用甲醇超声提取的方法制备供试品溶液,以乙腈-0.1%磷酸溶液梯度洗脱,流速:1 m L·min-1,柱温30℃,分段切换检测波长。结果建立了缩泉丸的HPLC指纹图谱共有模式,确立了14个共有峰,15批样品指纹图谱中各共有峰相对保留时间的相对标准偏差均<1.5%。结论该方法准确可靠、重复性好,建立的指纹图谱为缩泉丸质量评价提供依据。
