Integration of network pharmacology and bone marrow mesenchymal stem cells experimental research to reveal the molecular mechanisms for Hai Honghua medicinal liquor against osteoporosis

Background:Hai Honghua medicinal liquor(HHML)formula has been used in clinical practice for a long time,mainly for the treatment of freshly closed fractures,to promote osteogenesis,to increase bone mass,and thus to promote fracture healing.However,the underlying mechanisms of HHML in the treatment of osteoporosis(OP)are still unclear.Methods:Firstly,Traditional Chinese Medicines Systems Pharmacology Database and Analysis Platform and The Encyclopedia of Traditional Chinese Medicine were used to screen the targets of the active compounds of HHML.At the same time,OP targets were identified through GeneCards,Online Mendelian Inheritance in Man,DisGeNET,Therapeutic Target Database,Comparative Toxicogenomics Database and Human Phenotype Ontology databases.Next,protein-protein interaction and pathway networks were constructed for compound-disease common targets,and core targets and compounds were screened for molecular docking.Furthermore,rat bone marrow mesenchymal stem cells were extracted as model cells,and the osteogenic effects of HHML were verified via in vitro experiments.Results:Total of 343 common targets of HHML-OP and the top 10 target proteins in the protein-protein interaction network are TP53,AKT1,STAT3,HSP90AA1,ESR1,TNF,IL6,MAPK1,MAPK3 and MAPK8.Enrichment analysis yielded that the key molecular pathway was the PI3K/Akt signaling pathway.Molecular docking analysis showed that baicalein,erysodienone and naringenin docked with the target proteins AKT1,STAT3 and TP53,respectively,with low binding energy and high affinity.In addition,In vitro experiments demonstrated that HHML promoted the proliferation of bone marrow mesenchymal stem cells;compared with the control group,HHML-treated cells showed enhanced alkaline phosphatase staining,promoted the expression of OCN,RUNX2,BSP,and COL1 mRNAs as well as the expression of PI3K and AKT phosphorylated proteins.Secondly,the expression of target proteins revealed that HHML promoted the phosphorylation of STAT3 protein and inhibited the expression of P53.Conclusions:Our study investigated that HHML treatment with OP promotes bone formation possibly through activation of the PI3K/Akt signaling pathway and may involve STAT3 and TP53 target interactions.

Zuo Gui Wan Promotes Osteogenesis via PI3K/AKT Signaling Pathway:Network Pharmacology Analysis and Experimental Validation

Objective Osteogenesis is vitally important for bone defect repair,and Zuo Gui Wan(ZGW)is a classic prescription in traditional Chinese medicine(TCM)for strengthening bones.However,the specific mechanism by which ZGW regulates osteogenesis is still unclear.The current study is based on a network pharmacology analysis to explore the potential mechanism of ZGW in promoting osteogenesis.Methods A network pharmacology analysis followed by experimental validation was applied to explore the potential mechanisms of ZGW in promoting the osteogenesis of bone marrow mesenchymal stem cells(BMSCs).Results In total,487 no-repeat targets corresponding to the bioactive components of ZGW were screened,and 175 target genes in the intersection of ZGW and osteogenesis were obtained.And 28 core target genes were then obtained from a PPI network analysis.A GO functional enrichment analysis showed that the relevant biological processes mainly involve the cellular response to chemical stress,metal ions,and lipopolysaccharide.Additionally,KEGG pathway enrichment analysis revealed that multiple signaling pathways,including the phosphatidylinositol-3-kinase/protein kinase B(PI3K/AKT)signaling pathway,were associated with ZGW-promoted osteogensis.Further experimental validation showed that ZGW could increase alkaline phosphatase(ALP)activity as well as the mRNA and protein levels of ALP,osteocalcin(OCN),and runt related transcription factor 2(Runx 2).What’s more,Western blot analysis results showed that ZGW significantly increased the protein levels of p-PI3K and p-AKT,and the increases of these protein levels significantly receded after the addition of the PI3K inhibitor LY294002.Finally,the upregulated osteogenic-related indicators were also suppressed by the addition of LY294002.Conclusion ZGW promotes the osteogenesis of BMSCs via PI3K/AKT signaling pathway.

Fang-Xia-Dihuang decoction inhibits breast cancer progression induced by psychological stress via down-regulation of PI3K/AKT and JAK2/STAT3 pathways:An in vivo and a network pharmacology assessment

Background:The development and prognosis of breast cancer are intricately linked to psychological stress.In addition,depression is the most common psychological comorbidity among breast cancer survivors,and reportedly,Fang-Xia-Dihuang decoction(FXDH)can effectively manage depression in such patients.However,its pharmacological and molecular mechanisms remain obscure.Methods:Public databases were used for obtaining active components and related targets.Main active components were further verified by ultra-high-performance liquid chromatography-high-resolution mass spectrometry(UPLC-HRMS).Protein–protein interaction and enrichment analyses were taken to predict potential hub targets and related pathways.Molecule docking was used to understand the interactions between main compounds and hub targets.In addition,an animal model of breast cancer combined with depression was established to evaluate the intervention effect of FXDH and verify the pathways screened by network pharmacology.Results:174 active components of FXDH and 163 intersection targets of FXDH,breast cancer,and depression were identified.Quercetin,methyl ferulate,luteolin,ferulaldehyde,wogonin,and diincarvilone were identified as the principal active components of FXDH.Protein–protein interaction and KEGG enrichment analyses revealed that the phosphoinositide-3-kinase–protein kinase B(PI3K/AKT)and Janus kinase/signal transducer and activator of transcription(JAK2/STAT3)signaling pathways played a crucial role in mediating the efficacy of FXDH for inhibiting breast cancer progression induced by depression.In addition,in vivo experiments revealed that FXDH ameliorated depression-like behavior in mice and inhibited excessive tumor growth in mice with breast cancer and depression.FXDH treatment downregulated the expression of epinephrine,PI3K,AKT,STAT3,and JAK2 compared with the control treatment(p<0.05).Molecular docking verified the relationship between the six primary components of FXDH and the three most important targets,including phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha(PIK3CA),AKT,and STAT3.Conclusion:This study provides a scientific basis to support the clinical application of FXDH for improving depression-like behavior and inhibiting breast cancer progression promoted by chronic stress.The therapeutic effects FXDH may be closely related to the PI3K/AKT and JAK2/STAT3 pathways.This finding helps better understand the regulatory mechanisms underlying the efficacy of FXDH.

Research of the molecular mechanism of Hai Honghua medicinal liquor in the treatment of fracture based on network pharmacology combined with molecular docking

Background:Hai Honghua medicinal liquor(HHML),a famous hospital formula composed of 19 traditional Chinese medicines,has been successfully applied in treating soft tissue injury,fresh closed fracture,limb dysfunction after fracture healing,shoulder,neck and leg pain,knee joint pain and other clinical multiple diseases for 30 years in clinical.However,research on the material basis of HHML for the treatment of fracture healing-related disorders is still in a gap.Therefore,it is particularly important to explore the active ingredients,core targets and potential pharmacological mechanisms of HHML to promote fracture healing.Methods:We screened the core active components of each traditional Chinese medicine in formula and its action targets through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and Encyclopedia of Traditional Chinese Medicine database.The fracture related targets were retrieved from several different public databases,including GeneCards,Online Mendelian Inheritance in Man,DisGeNET and Therapeutic Target Database.Bioinformatics analysis to obtain key bioactive components,underlying targets and signaling pathways,containing the Venn diagram of the intersection with components and diseases gene targets,protein–protein interaction,as well as the Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes analysis,and finally molecular docking.Results:A total of 249 bioactive ingredients of HHML and 325 HHML-fracture-related targets were screened.The network analysis revealed that quercetin,luteolin,kaempferol,Licochalcone A,naringenin and 8-Isopentenyl-kaempferol may be potential candidate agents.Multiple targets are involved including TP53,MAPK3,STAT3,AKT1,MAPK1,HSP90AA1,ESR1 and PIK3CA may be closely linked targets.PI3K-AKT signaling pathway may play a significant role of HHML in treatment of fracture.What’s more,molecular docking suggested that 8-isopentenyl kaempferol,glycyrrhiza chalcone A,and naringenin bound to AKT1,PIK3CA,and ESR1,respectively,exhibiting lower energy and more stable characteristics.Conclusions:The findings indicate the potential active ingredients,target proteins and molecular mechanisms of HHML for the treatment of fractures to provide the exact idea for the next research on the mechanism of action of HHML formula for fracture treatment.

Simiao Wan alleviates obesity-associated insulin resistance via PKCε/IRS-1/PI3K/Akt signaling pathway based on network pharmacology analysis and experimental validation

Background:The purpose of the study was to investigatethe active ingredients and potential biochemicalmechanisms of Simiao Wan(SMW)in obesity-associated insulin resistance.Methods:An integrated network pharmacology method to screen the active compoundsand candidate targets,construct the protein-protein-interaction network,and ingredients-targets-pathways network was constructed for topological analysis to identify core targets and main ingredients.To find the possible signaling pathways,enrichment analysis was performed.Further,a model of insulin resistance in HL-7702 cells was established to verify the impact of SMW and the regulatory processes.Results:An overall of 63 active components and 151 candidate targets were obtained,in which flavonoids were the main ingredients.Enrichment analysis indicated that the PI3K-Akt signaling pathway was the potential pathway regulated by SMW in obesity-associated insulin resistance treatment.The result showed that SMW could significantly ameliorate insulin sensitivity,increase glucose synthesis and glucose utilization and reduce intracellular lipids accumulation in hepatocytes.Also,SMW inhibited diacylglycerols accumulation-induced PKCεactivity and decreased its translocation to the membrane.Conclusion:SMW ameliorated obesity-associated insulin resistance through PKCε/IRS-1/PI3K/Akt signaling axis in hepatocytes,providing a new strategy for metabolic disease treatment.

Elucidation of the hepatoprotective effect and mechanism of Melastoma dodecandrum Lour. based on network pharmacology and experimental validation

Objective:To systematically explore the effect and mechanism of melastomatis dodecandri herba(Melastoma dodecandrum Lour.)in the treatment of hepatitis based on network pharmacology.Method:We evaluated the hepatoprotective effects of M.dodecandrum in concanavalin A(Con A)-induced hepatitis in mice by assessing survival rate,histological analysis,serum transaminases,and related cytokines.Then the mechanism of action was predicted by a network pharmacology-based strategy.Based on the results,we measured the hepatic expression of related genes at mRNA level and proteins related to the phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)and nuclear factorkappa B(NF-кB)pathways.Results:Our study results clearly demonstrated that M.dodecandrum pretreatment significantly alleviated liver injury.This was demonstrated by an increase in survival rate,decreased severity of liver damage,and reduced serum transaminase levels compared with those in the Con A group.Moreover,M.dodecandrum significantly reduced the serum levels of tumor necrosis factor-a,interleukin-6,and interferon-g and increased the liver levels of superoxide dismutase,which indicated that M.dodecandrum exhibits anti-inflammatory and antioxidant activities.On the basis of network pharmacology,50 nodes were selected as major hubs based on their topological importance.Pathway enrichment analyses indicated that the putative targets of M.dodecandrum mostly participate in various pathways associated with the anti-inflammation response,which implies the underlying mechanism by which M.dodecandrum acts on hepatitis.Real-time fluorescent quantitative PCR analysis showed that M.dodecandrum downregulates the mRNA expression of interleukin-6,Toll-like receptor 7,interleukin-1 receptor-associated kinase-4,NF-кB and tumor necrosis factor-a in liver tissues.Western blotting showed that M.dodecandrum pretreatment protected against inflammation through activating the PI3K-Akt pathway by upregulating phosphorylated Akt(p-Akt)expression and suppressing NF-кB activation by inhibiting the phosphorylation of IKK,IkBa,and p65.Conclusion:The present work demonstrated the hepatoprotective effects of M.dodecandrum by regulating the PI3K/Akt and NF-кB pathways in Con A-induced mice,which provide insights into the treatment of hepatitis using M.dodecandrum.

Uncovering the mechanism of Sophora flavescens in the treatment of glioma based on network pharmacology

Glioma is a common primary intracranial tumor with high mortality and postoperative recurrence.Developing efficient therapies with lower toxicity is urgently needed.Sophora flavescens(SF)is a common Chinese medicine used to treat eczema,wet ulcers and itchy skin.Modern pharmacological studies have showed that SF has anti-glioma effects,but the mechanism of action remains unclear.This study aims to reveal the pharmacological mechanism of SF in treating glioma.The active components and related targets of SF were obtained from TCMSP.Genecard and Online Mendelian Inheritance in Man(OMIM)databases were used to explore the therapeutic targets for glioma.By making Venn diagram,we obtained 132 common targets of compounds and diseases.STRING databases and Cytoscape were used to construct diagrams of Protein-Protein Interaction(PPI)networks.Through the construction of PPI network,potential targets with degree value greater than the median were taken as core targets for further analysis.A total of 66 core targets were screened out.The degree values of TP53,HSP90AA1,MAPK1,and AKT1 were higher,indicating that these genes played important roles in this network.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were used to further discover the mechanism of active components in SF.Through enrichment analysis,it was found that the core targets were mainly enriched in PI3K/AKT signaling pathway,cell senescence related signaling pathway and IL-17 signaling pathway.In order to further explore the binding between active components and selected target,molecular docking was carried out.Finally,SwissADME was used to determine whether the compound could cross the blood-brain barrier.Based on network pharmacology,we speculated that matrine and formononetin in SF might inhibit PI3K/AKT signaling pathway and cellular senescence related signaling pathway by targeting AKT1,TP53,MAPK1 and other key targets.In summary,this study preliminarily explored the target and mode of action of SF in the treatment of glioma,laying the foundation for further research on its mechanism.

Mechanism of Pingyang Jiangya Formula in treating hypertension based on network pharmacology and in vivo study

Objective This study aimed to analyze the mechanism of action of the Pingyang Jiangya Formula(平阳降压方,PYJYF)in treating hypertension,based on network pharmacology,and to verify the subsequent predictions through animal experiments.Methods The active components and related target genes of PYJYF were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine(BATMAN-TCM),Encyclopedia of Traditional Chinese Medicine(ETCM),and Drug Bank databases and available literature.The hypertension target genes were screened based on Therapeutic Target Database(TTD),GeneCards,Online Mendelian Inheritance in Man(OMIM),UniProt,and relevant literature.The component-disease-target network intersection target genes were inputted into the STRING database,and the key target genes were selected according to the degree algorithm.Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were performed to explore the multitarget mechanism of action and molecular regulatory network of PYJYF in the treatment of hypertension.To verify this prediction,we used PYJYF to intervene in spontaneously hypertensive rats(SHRs)and Wistar–Kyoto rats(WKY)as normal control,and the noninvasive tail artery manometry method was used to measure systolic blood pressure(SBP)in the rat tail before PYJYF intervention.After drug intervention,the SBP of each group rats were measured and compared every week.Enzyme-linked immunosorbent assay(ELISA)was used to test plasma renin,angiotensin II(Ang II),and aldosterone(Ald)levels,and hematoxylin-eosin(HE)staining was used to observe pathological damage to the renal vessels in each group of rats.Western blot and reverse transcription real-time quantitative PCR(RT-PCR)were used to detect the protein and mRNA expression levels of PI3 K,AKT1,BAX,and Bcl-2,respectively.Results A total of 4123 hypertension targets were obtained from related databases.From the TCMSP and chemical databases,78 active components of PYJYF and the corresponding 401 drug targets were retrieved.Data analysis revealed that 208 drug targets directly interacted with the hypertension targets in PYJYF.The 10 targets most closely related to hypertension target proteins in PYJYF were directly retrieved from relevant databases.GO analysis revealed that 10 direct target proteins were involved in all aspects of the antihypertensive effects of PYJYF,as well as molecular biological processes,such as the regulation of blood pressure,renin-angiotensin-aldosterone system(RAAS),angiotensin-mediated ligand reactions,and biological stimulation of cardiomyocyte apoptosis.KEGG pathway enrichment analysis revealed that PYJYF directly affected 20 signaling pathways associated with hypertension.In animal experiments,PYJYF reduced the protein and m RNA levels of PI3 K,Akt,and Bax and upregulated the expression of the protein and m RNA levels of Bcl-2,reduced plasma renin,Ang II,and Ald levels,improved the hyperactivity of RAAS,and significantly reduced SBP in SHRs.Conclusion PYJYF is effective for hypertension therapy that acts through multiple compounds and targets.The possible underlying molecular mechanism includes regulating the PI3 K/Akt signaling pathway to suppress RAAS,increasing the ratio of Bcl-2/Bax proteins,and inhibiting apoptosis,thereby mediating the repair of renal and renal vascular damage caused by hypertension.These findings warrant further research for use in clinical settings.

基于关联规则与k均值聚类探讨黄褐斑外用方剂的药理规律

目的通过整理古籍中黄褐斑外用方剂,进而对现代临床中基于古籍的黄褐斑外用方药进行数据挖掘,获取其用药规律,探求其网络药理学依据,为临床治疗提供思路。方法检索中华医典数据库,获取古籍黄褐斑外用方药。搜索中国知网(CNKI)、万方数据库、维普中文期刊数据库(VIP)等,检索现代临床中黄褐斑的外用方药,筛选基于古方的外用方剂,建立黄褐斑外用方药数据库。使用中医传承计算平台(V3.0)分析药物关联规则,获取核心药物进行聚类分析。使用网络药理学探索核心药物潜在作用机制。结果整理得到古方37个,涉及中药78味;现代临床复方309个,涉及中药232味。现代临床中最常用的药物为白芷、茯苓、当归、白及、白附子等;最常用的药对为白芷-茯苓;高置信度药物关联规则有白及-白蔹->白芷、茯苓-白附子-僵蚕->白芷、茯苓-白及-僵蚕->白芷等;核心药物包括白芷、茯苓、僵蚕、白及、当归等12味中药。获取核心药物有效靶点347个,疾病靶点476个,药物与疾病交集靶点18个,京都基因和基因组数据库(KEGG)富集通路19条,并构建疾病-交集靶点-通路-核心药物-有效成分网络。结论核心药物对外用治疗黄褐斑在分子层面的作用机制存在依据,可为临床遣方用药提供依据,有助于提高中医外治黄褐斑的临床疗效。

原人参二醇型皂苷活性代谢物Compound K药理活性的研究进展

人参皂苷Compound K〔20-O-β-D-葡萄糖基-20(S)-原人参二醇,CK〕是原人参二醇型人参皂苷在人体内主要吸收形式和药效实体。近年来,CK的药理活性研究又有了新进展:①CK可通过活化胱天蛋白酶8直接或通过线粒体途径间接地激活胱天蛋白酶3,诱导肿瘤细胞凋亡;下调基质金属蛋白酶9基因的表达,抑制肿瘤细胞的转移;②可通过下调多种炎症因子如白细胞介素-4(IL-4),IL-6,肿瘤坏死因子α等及环氧合酶2与一氧化氮合酶的表达而发挥抗炎、抗过敏和神经保护作用;③可阻断ATP敏感性K+离子通道,促进胰岛素分泌,增强组织胰岛素敏感性,抑制糖异生,促进糖酵解,对胰岛素依赖性糖尿病表现出良好的疗效;④可上调核苷切除修复相关蛋白基因的表达,减少紫外线引起的DNA损伤,防止皮肤老化等。本文重点对近几年有关CK的药理活性及其作用机制研究新进展进行综述。

安胃汤含药血清通过调节PI3K/Akt信号通路促进MC细胞凋亡的研究

目的：探讨安胃汤含药血清对MC细胞（MNNG诱导人胃黏膜上皮细胞系（GES-1）恶性转化）PI3K/Akt信号通路相关因子及细胞凋亡影响。方法：根据中药复方血清药理学实验方法,以MC细胞为研究对象,设立空白组（A组）、安胃汤组（B组）、安胃汤＋阻断剂组（C组）、阻断剂组（D组）。含药血清处理12、24、48 h后的各组细胞采用流式细胞术检测细胞凋亡率;Western Blot和Real-time PCR检测PI3Kα、AKT、XIAP蛋白及mRNA的表达。结果：A组细胞凋亡率比较低,B、C、D组细胞凋亡率有所增加,C组细胞凋亡率增加最为明显;与A组相比,B、C、D 3组细胞内PI3Kα、AKT、XIAP基因及蛋白表达有所降低（P〈0.01或P〈0.05）。结论：安胃汤能促进MC细胞细胞凋亡,可能与其通过调控PI3K/Akt通路,下调XIAP基因及蛋白有关。

K_(ATP)通道介导的油茶皂甙对在体大鼠心肌产生的缺血预适应样保护作用

目的 :研究油茶皂甙 (SQS)对大鼠心肌产生的缺血预适应样保护作用及与KATP通道的关系。方法 :以ISO诱发大鼠心肌缺血损伤为模型 ,使用KATP通道特异性阻断剂gliberclamide (GLI 5mg·kg-1)。实验分为四组 ,分别为生理盐水对照组 (NS组 )、ISO诱发损伤组 (I R组 )、SQS组 (I R +SQS 0 .2mg·kg-1)和GLI组 (I R +GLI +SQS)。在注射ISO前 ,从阴茎静脉预适应注射各被试药物或NS ,每天 1次 ,连续 3d。末次给药后立即皮下多点注射ISO。NS组皮下注射等量NS。分别测定大鼠ECG及末次给药后血清CPK活性 ,FFA和腺苷含量。结果 :预适应ivSQS能有效地保护ISO所致大鼠心肌损伤 ;先ivGLI后再给予SQS ,则SQS对心肌损伤的保护作用明显减弱。结论 :SQS对ISO所致心肌缺血大鼠可产生药理性预适应保护作用 。

人参皂苷CK的研究进展

人参皂苷coumpound K(CK)是人参中原人参二醇型皂苷在人体肠道内的主要代谢产物,属于稀有人参皂苷。人参皂苷CK独特的生物活性已经引起了人们广泛关注,针对它的科学研究也日益增多。因此,有必要介绍近年来人参皂苷CK药理活性和制备方法方面的研究进展。

苦参碱对K-ras基因突变型结肠癌细胞株增殖的抑制作用及机制研究

【目的】观察苦参碱对K-ras基因突变型结肠癌SW480细胞增殖的抑制作用及机制。【方法】以不同浓度苦参碱干预K-ras基因突变型人结肠癌SW480细胞,采用新型四唑氮盐（MTS）做比色分析测定细胞增殖,流式细胞技术（FCM）检测细胞凋亡,细胞划痕法观察细胞侵袭转移能力的变化,Western blotting方法检测细胞内丝裂原活化蛋白激酶（MAPK）通路下游关键激酶MEK1/2蛋白的表达。【结果】与空白对照组比较,0.125～1 mg/m L浓度的苦参碱能显著抑制SW480细胞增殖,促进细胞凋亡,有效抑制结肠癌细胞的移行,降低MEK1/2蛋白的表达（P〈0.05）,其作用强度随药物浓度增加有升高的趋势。【结论】苦参碱可能通过下调MAPK信号通路下游MEK1/2蛋白的表达,有效抑制SW480细胞的增殖和移行,并促进其凋亡。

补肾通络方对去卵巢大鼠破骨细胞组织蛋白酶K表达的影响

【目的】探讨组织蛋白酶K(cathepsin K,CK)在骨质疏松症发病中的意义,观察补肾通络方对去卵巢大鼠破骨细胞CK表达的影响。【方法】选用30只雌性SD大鼠随机分成3组:假手术组、模型组及补肾通络方组(中药组,剂量为12.46 g.kg-1.d-1);后2组切除双侧卵巢,4周后中药组给予补肾通络方灌胃。分别于第6、12周测定各组大鼠全身骨密度(BMD),并采用Western blot及逆转录—聚合酶链反应(RT-PCR)法测定体外分离培养破骨细胞中CK的蛋白及基因表达。【结果】体外分离培养椎骨来源的破骨细胞具有成熟破骨细胞的表型特征,去卵巢模型大鼠骨密度在时效上与CK的表达呈现负相关;补肾通络方以时效方式下调CK蛋白及CK mRNA的表达水平(均P<0.05)。【结论】CK的过度表达是导致骨密度下降的重要原因,是骨质疏松症发生的潜在危险因子;以时效方式下调CK蛋白及CK mRNA的表达进而治疗骨质疏松症可能是补肾通络方的作用机制之一。

The potential mechanism of Isodon suzhouensis against COVID-19 via EGFR/TLR4 pathways

Corona Virus Disease 2019(COVID-19)has brought the new challenges to scientific research.Isodon suzhouensis has good anti-inflammatory and antioxidant stress effects,which is considered as a potential treatment for COVID-19.The possibility for the treatment of COVID-19 with I.suzhouensis and its potential mechanism of action were explored by employing molecular docking and network pharmacology.Network pharmacology and molecular docking were used to screen drug targets,and lipopolysaccharide(LPS)induced RAW264.7 and NR8383 cells inflammation model was used for experimental verification.Collectively a total of 209 possible linkages against 18 chemical components from I.suzhouensis and 1194 COVID-19 related targets were selected.Among these,164 common targets were obtained from the intersection of I.suzhouensis and COVID-19.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enriched 582 function targets and 87 target proteins pathways,respectively.The results from molecular docking studies revealed that rutin,vitexin,isoquercitrin and quercetin had significant binding ability with 3 chymotrypsin like protease(3CLpro)and angiotensin converting enzyme 2(ACE2).In vitro studies showed that I.suzhouensis extract(ISE)may inhibit the activation of PI3K/Akt pathway and the expression level of downstream proinflammatory factors by inhibiting the activation of epidermal growth factor receptor(EGFR)in RAW264.7 cells induced by LPS.In addition,ISE was able to inhibit the activation of TLR4/NF-κB signaling pathway in NR8383 cells exposed to LPS.Overall,the network pharmacology and in vitro studies conclude that active components from I.suzhouensis have strong therapeutic potential against COVID-19 through multi-target,multi-pathway dimensions and can be a promising candidate against COVID-19.

加减补肺汤对COPD肺气虚证大鼠血浆中6-K-PGF1α TXB2含量的影响

采用烟熏复合木瓜蛋白酶雾化吸入法复制Wistav大鼠COPD"肺气虚证"模型。并给予加减补肺汤治疗,观察大鼠血浆中6-K-PGF1α、TXB2含量,探讨其作用机理。研究结果显示,加减补肺汤能显著降低TXB2、提高6-K-PGF1α含量,减轻呼吸道的损伤,对促进机体和肺组织功能状态的恢复有一定的作用。

维生素K_3对安定血药浓度及药理作用的影响

用动物实验研究了维生素Ｋ３对安定血药浓度及药理作用的影响。结果表明，维生素Ｋ３能明显缩短安定血药浓度分布半衰期和消除半衰期，提高安定体内清除率。增加家兔呼吸频率，但不影响麻醉后家兔的心电图、血压和心率。能缩短安定所致小鼠睡眠时间，具有催醒作用。

Anti-tumor activity of Sanguisorba officinalis L.in non-small cell lung cancer and induced apoptosis via PI3K/Akt/mTOR signaling pathway

OBJECTIVE To investigate the pharmacological effect and mechanism of Sanguisorba officinalis L.(SOL)in non-small cell lung cancer(NSCLC)in vitro and in vivo based on network pharmacology.METHODS Network pharmacology was used to analyze the improving effect of SOL on NSCLC and possible targets.Cell counting kit 8(CCK-8)and 5-ethynyl-2′-deoxyuridine(EdU)staining,Western blotting,flow cytometry of AnnexinⅤ/PI,Hoechst 33342/PI staining detection and immunofluorescence were utilized in vitro.H&E staining,immunohistochemistry staining and Western blotting were performed in vivo.RESULTS Based on network prediction,we analyzed the 208 common targets of SOL and NSCLC.36 core targets in 208 common targets were obtained through cytoscape analysis.And the top 10 core targets included Akt,mTOR,EGFR,etc..KEGG analysis showed that PI3K-Akt signaling pathway was the most likely pathway.Furthermore,the mechanism study found that SOL could activate the PI3K/Akt/mTOR signaling pathway in vitro and in vivo.The anti-proliferative effect of SOL in A549 and H1299 cells was measured and validated by CCK-8 and EdU assay.Immunohistochemical results of Ki67 showed that SOL effectively inhibited tumor growth in vivo.SOL also significantly inhibited the migration and invasion of A549 and H1299 cells.SOL significantly increased the percentage of cells with PI signal in A549 and H1299,and the process of cell death of A549 cells indicated that SOL induced apoptosis.The PARP-1 and caspase-3 in A549 and H1299 were found to be activated in a dose manner.The results in vivo were consistent with those in vitro.CONCLUSION SOL-induced,caspase-3-mediated apoptosis via the induction of the PI3K/Akt/mTOR signaling pathway in NSCLC,which further clarified the mechanism of SOL in the inhibition of NSCLC,and thereby provided a possibility for SOL to serve as a novel therapeutic agent for NSCLC in the future.

临床医学专业《药理学》试卷分析和反思

目的分析某地方高校临床医学专业本科五年制药理学期末考试试卷。方法2022年7月,收集临床医学2020级药理学期末试卷142份,有效试卷140份,其中,临床医学2020级1班66份,临床医学2020级2班74份。以教育测量学原理分析试卷难度、区分度、信度和效度,运用数据挖掘算法中主成分分析和K-均值聚类分析学生考试成绩。结果试卷总体难度0.58,区分度0.34,信度0.80。全部学生平均成绩(58.2±13.9)分,成绩符合正态分布,两班级学生成绩差异无统计学意义(P>0.05)。主成分分析发现影响试卷成绩的两个维度,不同班级又有所差异;K-均值聚类依据考核知识点划分不同层次的学生。结论试卷具有分析价值。不同班级学生对药理学各章节知识掌握情况存在差异,课程教学效果亟待改进。本次试卷分析为后续的改进课程教学和提高人才培养质量提供实证。