Revealing mechanism of Methazolamide for treatment of ankylosing spondylitis based on network pharmacology and GSEA

Abstract Methazolamide is a carbonic anhydrase (CA) inhibitor with satisfactory safety. Our previous studies have demonstrated the elevation of CA1 expression and the therapeutic effect of Methazolamide in Ankylosing spondylitis (AS). In this study, we explored the pathogenic role of CA1 and the pharmacological mechanism of Methazolamide in AS through Gene Set Enrichment Analysis (GSEA) and network pharmacology. Seven out of twelve CA1 related gene sets were enriched in AS group. CA1 was core enriched in above seven gene sets involving zinc ion binding, arylesterase activity and one carbon metabolic process. Functional analysis of the candidate target genes obtained from the intersection of AS associated genes and Methazolamide target genes indicated that Methazolamide exerts therapeutic effects on AS mainly through inflammatory pathways which regulate the production of tumor necrosis factor, IL-6 and nitric oxide. PTGS2, ESR1, GSK3β, JAK2, NOS2 and CA1 were selected as therapeutic targets of Methazolamide in AS. Molecular docking and molecular dynamics simulations were performed successfully. In addition, we innovatively obtained the intersection of Gene Ontology (GO)/Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses and GSEA results, and found that 18 GO terms and 5 KEGG terms were indicated in the pharmacological mechanism of Methazolamide in AS, involving bone mineralization, angiogenesis, inflammation, and chemokine signaling pathways. Nevertheless, validation for these mechanisms is needed in vivo/vitro experiments

Retrospective Evaluation of New Chinese Diagnostic Scoring System for Disseminated Intravascular Coagulation.

To retrospectively validate the new Chinese DIC scoring system (CDSS).This study retrospectively collected the information of 619 patients (371 cases with non-hematologic malignancies, 248 cases with hematologic malignancies) who suspected of DIC in Wuhan Union Hospital during 2013-4 to 2014-6. We validated CDSS by comparing it with three leading scoring systems, from International Society on Thrombosis and Haemostasis (ISTH), Japanese Association for Acute Medicine (JAAM) and Japanese Ministry of Health and Welfare (JMHW), and evaluated its prognostic value by 28 days mortality, APACHE II and SOFA score.In non-hematologic malignancies, CDSS was more specific than JAAM (72.55% vs. 50.49%, p<0.05) and more sensitive than ISTH (77.07% vs. 62.03%, p<0.05). In hematologic malignancies, the area under the ROC curve of CDSS was larger than ISTH and JMHW (0.933 vs. 0.889, p<0.01 with ISTH, 0.944 vs. 0.845, p<0.01 with JMHW). In addition, the 28-day mortality rate, SOFA scores, APACHE II scores of DIC patients diagnosed by CDSS were significantly greater than non-DIC (P <0.05).We are the first group to propose CDSS. It emphasized the values of the clinical manifestations, the rapidly declining platelet count, APTT in the diagnosis of DIC and used D-dimer as the fibrin-related maker. DIC with hematological malignancies was treated as a special part. In this study we can see that CDSS displayed an acceptable property for the diagnosis of DIC with appropriate sensitivity and specificity, and also had a good prognostic value for DIC patients

The flow diagram of the whole study process.

<p>The flow diagram of the whole study process.</p

Chinese DIC Scoring System (CDSS).

<p>Diagnosis: Seven points or more.</p><p>*If underlying disease is hematological malignancy:1.ignore item(1) from Clinical manifestations; 2.Platelet count: PLT<50×10⁹/l:1point,≥50% decrease within 24 hrs:1point; 3.Diagnosis: Six points or more. PT, prothrombin time. APTT, activated partial thromboplastin time.</p><p>Chinese DIC Scoring System (CDSS).</p

Characteristics of the diagnosis and mortality by each criterion in non- hematological malignancy.

<p>ISTH, International Society on Thrombosis and Haemostasis; JMHW, Japanese Ministry of Health and Welfare; JAAM, Japanese Association for Acute Medicine; CDSS, Chinese DIC scoring system; DIC, disseminated intravascular coagulation; AUC, area under receiver operating characteristic curve; OR, Odds ratio; CI, confidence interval.</p><p>a:p<0.05 for CDSS compared with other criteria;</p><p>b:p>0.05 for CDSS compared with other criteria;</p><p>c:P>0.05 between JAAM and ISTH;</p><p>*P>0.05 between any two;</p><p>#p<0.05 for each OR.</p><p>Characteristics of the diagnosis and mortality by each criterion in non- hematological malignancy.</p

Characteristics of DIC and non-DIC patients diagnosed by CDSS.

<p>*: sort order (infection, solid tumors, autoimmune diseases, trauma/post-surgery/poisoning, obstetric complications, vascular anomalies, shock/hypoxic-ischemic, severe liver disease, unknown reason, hematological malignancy); CDSS, Chinese DIC scoring system; DIC, disseminated intravascular coagulation; APACHE, Acute Physiology and Chronic Health Evaluation; SOFA, Sequential Organ Failure Assessment; PLT, Platelet counts; APTT, activated partial thromboplastin time; PT, Prothrombin time; FDP, fibrin/fibrinogen degradation products; ATA, Antithrombotic Agents.</p><p>Characteristics of DIC and non-DIC patients diagnosed by CDSS.</p

Acute Physiology and Chronic Health Evaluation II (APACHE II) score and 28-day mortality by the clinical manifestation.

<p>Bar graph shows 28-day mortality (*p = 0.000). Line graph shows the average APACHE II score (**p = 0.000).</p

Classification of underlying diseases and diagnostic rate by each criteria for DIC.

<p>DIC, disseminated intravascular coagulation; ISTH, International Society on Thrombosis and Haemostasis; JMHW, Japanese Ministry of Health and Welfare; JAAM, Japanese Association for Acute Medicine; CDSS, Chinese DIC scoring system.</p><p>Classification of underlying diseases and diagnostic rate by each criteria for DIC.</p

Acute Physiology and Chronic Health Evaluation II (APACHE II) score and 28-day mortality by prolongation of activated partial thromboplastin time (APTT).

<p>Bar graph shows 28-day mortality (*p = 0.000). Line graph shows the average APACHE II score (**p = 0.000).</p

Distribution of patients with hematological malignancy according to the three diagnostic criteria.

<p>Left, comparison between the International Society on Thrombosis and Haemostasis (ISTH) and Chinese DIC scoring system (CDSS); right, comparison between the Japanese Ministry of Health and Welfare (JMHW) and Chinese DIC scoring system (CDSS). Numbers in parentheses are of nonsurvivors.</p