scholarly journals RELATO DE CASO: PACIENTE COM PERMANÊNCIA DE IGM POSITIVO PARA COVID-19

2021 ◽  
Vol 16 (2) ◽  
pp. 22-24
Author(s):  
Julyana Alves Barreto Rodrigues ◽  
Bianca Viana de Araújo Henriques
Keyword(s):  
Diabetes Mellitus ◽  
Interferon Beta ◽  
De Novo ◽  
Rt Pcr

Introdução: A Covid-19 doença causada pelo coronavírus, teve seu primeiro caso relatado em dezembro de 2019, em Wuhan na China. Atualmente já vitimou cerca de 961.656 óbitos no mundo e ainda segue ameaçando a saúde global. O SARS-CoV-2 pertence ao gênero betaCovs, da subfamília Orthocoronavirinae, da família Coronaviridae. É um vírus envelopado (ou seja, há uma membrana lipídica de formato duplo e também proteínas inseridas) de fita única de RNA, em forma de coroa, devido as glicoproteínas em forma de espinhos, presentes em seu envelope. A sua entrada na célula hospedeira humana se dá a partir do auxílio da glicoproteína S se ligar a ECA II (enzima conversora de angiotensina) da célula hospedeira e a usa-la como aparato para replicação (CASCELLA et al., 2020; VELAVAN et al., 2020). Nos dias atuais, é possível descrever fatores de risco a partir de grupos de risco que obtiveram uma evolução desfavorável tais como indivíduos com idade superior a 60 anos, portadores de doenças crônicas, como exemplo diabetes mellitus, hipertensão arterial sistêmica e/ou doenças pulmonares crônicas, tabagista de longa data como também os que fazem uso de quimioterápicos, corticosteroides de uso prolongado ou imunobiológicos. Para a definição do diagnóstico de COVID-19 é preciso somar as informações clínico-epidemiológicas com exames RT-PCR e/ou sorologia quando possível e o exame de imagem com a tomografia computadorizada. RT-PCR é considerado o método padrão ouro apresenta uma sensibilidade em torno de 63% quanto aos testes sorológicos tem maior sensibilidade porém somente após 7-9 dias de sintomas. Atualmente, vários medicamentos veem sendo estudados para o tratamento clinico da Covid 19 dentre eles pode-se destacar: Remdesivir, Ritonavir, Pirfenidona, Cloroquina/Hidroxicloroquina, Interferon-beta, Sarilumab. Entretanto, sabe-se hoje que a dexametasona foi o único medicamento que reduziu a mortalidade em pacientes hospitalizados Objetivos: Entender a fisiopatologia viral e conhecer os principais fatores de risco da doença. Descrição do caso: D.R.A, 27 anos, sexo masculino, hipertenso controlado, iniciou no dia 19/07 com quadro de dor de garganta, espirros, obstrução nasal e coriza. No dia seguinte apresentou fraqueza e dor no corpo. No dia 22/07 os sintomas se agravaram e o paciente começou a apresentar tosse persistente e dor nas costas, na região torácica, irradiando para nuca, sendo potencializada com o esforço e atenuava com o repouso, então realizou o Teste PCR para COVID – 19, testando positivo, no mesmo dia. No dia 23/07, iniciou quadro de febre alta de 38°C, associada a calafrios, que permaneceu por 4 dias. Procurou assistência médica no mesmo dia que iniciou a febre e foi prescrito hidroxicloroquina e Azitromicina por 1 semana e Prednisona por 2 semanas. No dia 05/08 já não apresentava mais sintomas, com exceção da tosse e realizou novo teste, desta vez o teste rápido, dando positivo para IgM e IgG. O paciente continuou com tosse persistente, e no dia 28/08, 37 dias depois do início dos sintomas, realizou novo teste rápido, continuando a apresentar IgM e IgG positivos. Passou por avaliação médica e foi prescrito Moxifloxacino 400mg, Alegra e Acetilcisteína xarope por mais 14 dias. Por volta do 10º dia de medicação a tosse desapareceu definitivamente. Conclusão: Através deste relato é possível observar que não há um tempo preciso para a negativação do IgM em pacientes que tiveram a COVID-19, mesmo já apresentando IgG positivo há algum tempo. Além disso, ainda podemos perceber que o tratamento, por vezes, não é adequado para a condição do paciente, visto que o mesmo permanece com alguns sintomas mesmo após o tratamento e período ativo da doença. Casos em que o tratamento clínico não foi suficiente, devem ser reavaliados para verificar a necessidade de novo ciclo de medicação.

Immunsuppression nach Nierentransplantation

2020 ◽  
Vol 24 (03) ◽  
pp. 103-103
Author(s):  
Volker Aßfalg
Keyword(s):  
Diabetes Mellitus ◽  
De Novo ◽  
Onset Diabetes ◽  
Ciclosporin A ◽  
Unerwünschte Nebenwirkungen ◽  
New Onset

Der Goldstandard der Immunsuppression nach Nierentransplantation gemäß aktuellen KDIGO-Empfehlungen 1 besteht nach wie vor aus einem Calcineurininhibitor (CNI), Mycophenolsäure und Steroiden – der sog. Tripel-Therapie. Der große Durchbruch in der Langzeitüberlebensrate von Nierentransplantaten gelang erst in den 1990er-Jahren mit dem Einsatz von Ciclosporin A. Mit Einführung des ähnlich wirkenden, aber potenteren Tacrolimus 2 wurde dieser CNI in die Empfehlungen der KDIGO als Erstlinienpräparat in der de novo Immunsuppression aufgenommen 1. Vonseiten des Nebenwirkungsprofils zeigen die CNI jedoch unerwünschte Nebenwirkungen wie z. B. Nephrotoxizität, die im Rahmen der sog. CNI-Toxizität die Transplantatlangzeitfunktion einschränken und limitieren kann. Darüber hinaus findet sich ein erhöhtes Risiko für Hypertonie, Fettstoffwechselstörungen und insbesondere für Tacrolimus die Auslösung eines Post-Transplantations-Diabetes (NODAT: New Onset Diabetes After Transplantation) oder Aggravierung eines bestehenden Diabetes mellitus.

Genexpressionsmuster von fibrinolytischen Faktoren des Urokinase-Plasmin-Systems bei Dermatoliposklerose

Phlebologie ◽  
1999 ◽  
Vol 28 (01) ◽  
pp. 1-6 ◽  
Author(s):  
Ch. Stetter ◽  
E. Schöpf ◽  
J. Norgauer ◽  
W. Vanscheidt ◽  
Y. Herouy
Keyword(s):  
Mrna Expression ◽  
De Novo ◽  
Ulcus Cruris ◽  
Ulcus Cruris Venosum ◽  
Rt Pcr ◽  
Fand Sich ◽  
Pai 1

ZusammenfassungDie Dermatoliposklerose (DLS) entwickelt sich als Folge einer progredienten primären Varikosis oder eines postthrombotischen Syndroms (PTS). Trotz bestehender Hinweise auf eine veränderte intravasale fibrinolytische Aktivität bei der chronisch-venösen Insuffizienz (CVI), wurden bisher fibrinolytische Faktoren im perivaskulären Gewebe nicht untersucht. Kürzlich zeigten wir, daß bei Dermatoliposklerose Matrix-Metalloproteinasen exprimiert und aktiviert werden. Da spezifische fibrinolytische Faktoren wichtige Haupteffektoren der Matrix-Metalloproteinasenaktivierung sind, untersuchten wir kürzlich die Genexpression der Plasminogenaktivatoren vom Urokinasetyp (uPA) und vom Gewebetyp (tPA), des Urokinase-Rezeptor (uPA-R) sowie der Plasminogenaktivator-Inhibitoren (PAI-1 und PAI-2) in Gewebsbiopsien von Patienten mit Dermatoliposklerose. Zum Nachweis verwandten wir dabei die Technik der reversen Transkription und Polymerase-Kettenreaktion (RT-PCR). Es fand sich in allen Hautproben (n = 21) eine signifikant erhöhte mRNA-Expression von uPA und uPA-R im Vergleich zu gesunder Haut (n = 12). Dagegen konnte kein signifikanter Unterschied für mRNA-Transkripte von tPA, PAI-1 und PAI-2 nachgewiesen werden. Die Dermatoliposklerose zeichnet sich somit durch erhöhte transkriptionelle Expression von uPA und uPA-R aus. Eine gesteigerte De-novo-Synthese von uPA und uPA-R könnte daher bei der Aktivierung von Matrix-Metalloproteinasen und entsprechend in der Pathogenese des Ulcus cruris venosum eine zentrale Rolle spielen.

Identification of α-enolase as a prognostic and diagnostic precancer biomarker in oral submucous fibrosis

2017 ◽  
Vol 71 (3) ◽  
pp. 228-238 ◽  
Author(s):  
Swarnendu Bag ◽  
Debabrata Dutta ◽  
Amrita Chaudhary ◽  
Bidhan Chandra Sing ◽  
Mousumi Pal ◽  
...  
Keyword(s):  
De Novo ◽  
Pain Treatment ◽  
Oral Submucous Fibrosis ◽  
Peptide Sequencing ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Sequence Coverage ◽  
Protein Marker ◽  
Peptide Mass ◽  
Submucous Fibrosis

AimsDiagnostic ambiguities regarding the malignant potentiality of oral submucous fibrosis (OSF), an oral precancerous condition having dysplastic and non-dysplastic isoforms are the major failure for early intervention of oral squamous cell carcinoma (OSCC) patients. Our goal is to identify proteomic signatures from biopsies that can be used as precancer diagnostic marker for patient suffering from OSF.MethodsThe high throughput techniques adopting de novo peptide sequencing (1D SDS-PAGE coupled nanoLC MALDI tandem mass spectrometry (MS/MS)-based peptide mass fingerprint), immunohistochemistry (IHC), Western blot (WB) and real-time PCR (RT-PCR) analysis are considered for such biomarker identification and multilevel validations.ResultsAlpha-enolase is identified as an overexpressed protein in biopsies of oral submucous fibrosis with dysplasia (OSFWD) compared with oral submucous fibrosis without dysplasia (OSFWT) and normal oral mucosa (NOM). Total proteome analysis of an overexpressed protein band around 47 kDa of OSFWD identifies 334 peptides corresponding to 61 human proteins. Among them α-enolase is identified as a prime protein with highest number of peptides (44 out of 334 peptides) and sequence coverage (66.4%). Furthermore, RT-PCR, WB and IHC analysis also show mRNA and tissue level upregulation of α-enolase in OSFWD validating α-enolase as precancer marker.ConclusionsThis study for the first time identifies and validates α-enolase as a novel biomarker for early diagnosis of malignant potentiality of OSF. Hence, the identified protein marker, α-enolase can help in early therapeutic intervention of OSF patients leading to the reduction of patient’s pain, treatment cost and enhancement of patient’s quality of life.

scholarly journals Effects of Momordica Charantia Silver Nanoparticles on the expressions of Genes Associated With Lipid Metabolism and Nephrotoxicity in Streptozotocin-Induced Rats

2021 ◽  
Vol 37 (2) ◽  
pp. 126-133
Author(s):  
O.O Elekofehinti ◽  
M.O Akinjiyan
Keyword(s):  
Diabetes Mellitus ◽  
Silver Nanoparticles ◽  
Lipid Metabolism ◽  
Silver Nitrate ◽  
Nitrate Solution ◽  
Diabetic Control ◽  
Untreated Group ◽  
Rt Pcr ◽  
Hmg Coa Reductase ◽  
Coa Reductase

Hyperlipidemia and hyperglycemia have been implicated in diabetes mellitus (DM) leading to complications such as nephropathy. Medicinal plants like Mormodica charantia (MC) have been used in the treatment of DM over the years but little is known about their mechanisms of action. This study used biotechnology tools to investigate and compare the effects of M. charantia silver nanoparticles (MCSNPs) with M. charantia extract on expressions of genes linked with nephrotoxicity, lipid and glucose metabolisms using reverse-transcriptase polymerase chain reaction (RT-PCR) in streptozotocin-induced diabetic rats. The genes investigated include kidney injury molecule-1 (KIM-1), 3-hydroxyl, 3-methyl glutaryl_coA reductase (HMG-CoA reductase), peroxisome proliferator-activated receptor alpha and gamma (PPARα and PPARγ). Synthesis of MCSNPs was done using 1 mM concentration of aqueous silver nitrate solution at ratio 1:9 (v/v). Experimental rats were induced intraperitoneally with streptozotocin (65 mg/kg) and divided into six groups viz: diabetic control; normal control; silver nitrate (10 mg/kg); MCSNPs (50 mg/kg); Metformin (100 mg/kg) and M. charantia fraction (100 mg/kg). Sacrifice was done after 12 days of treatment and RT-PCR was then used to investigate gene expressions in liver and kidney tissues of the rats. The expression of HMG-CoA reductase gene was significantly upregulated (p<0.05) upon treatment with 50 mg/kg MCSNPs relative to the diabetic untreated group. M. charantia extracts and MCSNPs significantly upregulate (p<0.05) the expressions of PPAR-α and PPAR-γ compared to the diabetic control. Also, a significant (p<0.05) down-regulation of KIM-1 mRNA expression was observed in MCSNPs- treated group, relative to the diabetes untreated group. M. charantia silver nanoparticles could be a potent antidiabetic agent due to its potential to modulate genes associated with lipid metabolism and nephrotoxicity. Keywords: Medicinal plant; Diabetes Mellitus; Silver Nanoparticles; nephrotoxicity; gene expression

scholarly journals Cucurbit chlorotic yellows virus infecting Rehmannia glutinosa was detected in China

Plant Disease ◽  
2021 ◽  
Author(s):  
Kun Zhang ◽  
Xinjian Zhuang ◽  
Xiao Guo ◽  
Hongmei Xu ◽  
Zhen He ◽  
...  
Keyword(s):  
De Novo ◽  
Viral Disease ◽  
Henan Province ◽  
Plant Sample ◽  
Herbaceous Plant ◽  
Rehmannia Glutinosa ◽  
Rt Pcr ◽  
Illumina Hiseq ◽  
Two Samples ◽  
Cucurbit Chlorotic Yellows Virus

Rehmannia glutinosa Libosch. is a perennial herbaceous plant of the family Scrophulariaceae. Its roots can be used as traditional Chinese medicine. The asexual reproduction by vegetative organ of R. glutinosa lead to an increased viral disease that seriously affects its yield and quality (Kwak et al. 2020; Kwak et al. 2018; Ling and Liu 2009). Leaves of R. glutinosa in Wenxian County, Henan Province, China showed symptoms of chlorosis, mosaic and irregular yellow in August 2019. In general, the older leaves at the base or middle of the plant (sample 2# and 5#) first became irregular yellowing, followed by a gradual extend to the leaves at the top (Supplementary Fig. S1A). Six plants (2#, 3#, 5#, 7#, 8#, and 9#) with these symptoms were collected. The total RNA was extracted and its siRNAs were obtained. High-throughput siRNA sequencing (Sangon, Shanghai, China) was performed on Illumina Hiseq 2000 platform with paired-end method after siRNA library construction (NEBNext Ultra II RNA Library Prep Kit, NEB, UK). Sequencing files were treated with Illumina’s CASAVA pipeline (version 1.8). The length of the resulting reads with adaptor removed were mostly distributed ranging from 21-24 nt (Supplementary Fig. S1B). The Velvet Software 0.7.31 (k=17) was taken to do de novo assembling, and the contigs (∼13,000, Contigs > 300 bp) were used to perform BLASTN against GenBank database. Two viruses, Rehmannia mosaic virus (ReMV) and cucurbit chlorotic yellows virus (CCYV), were frequently appeared in analyzed six symptomatic samples. To further identify the infection of CCYV to R. glutinosa, ten samples with virus-infected symptoms were randomly collected. Total protein and RNAs were extracted for RT-PCR and ELISA (HALING. Shanghai, China). A specific pair of primers (Supplementary Table S1) were designed to amplify the 753-bp length coat protein (CP) gene of CCYV. The result showed that two samples appeared a specific band of expected size on the agarose gel, which indicated that they were infected by CCYV (Supplementary Fig. S1C, Upper panel). The same result was obtained by ELISA assay (Supplementary Fig. S1D). The amplified CP fragment of CCYV was recycled and purified by TIANgel Midi Purification Kit (Tiangen, Beijing, China), followed by cloned into pMD19-T (TaKaRa, Dalian, China) and transformed into E. coli DH5a.Ten separate clones were selected and sequenced (Sangon, Shanghai, China) after PCR verification. The obtained sequences (GenBank accession No. MW521380 & MW521381) were analyzed by BLASTN and bioEdit software (version 7.2.3). The results showed 100% identity with the CCYV CP sequences that mainly derived from infected cucurbit. To confirm the occurrence and distribution of CCYV and ReMV in planting area, the other twenty-four samples (20 with chlorosis and stunt symptoms and 4 with invisible symptoms) were randomly collected for RT-PCR in different regions of Henan Province (Supplementary Table S1). The results showed that the CCYV and ReMV infection rate were 20.5% and 61.7%, respectively. Co-infection of the CCYV and ReMV was 5.8% in fields (Supplementary Table S2). In sum, these results indicated the CCYV can naturally infect R. glutinosa in China. CCYV is transmitted by white-fly in a semi-persistent manner and mainly damages cucurbits (Orfanidou et al. 2017). CCYV has been discovered in many places (Huang et al. 2010). To date, there is no report about CCYV infecting R. glutinosa in nature. This is the first report of CCYV naturally infect R. glutinosa in China.

Interrupção da gestação em paciente com infecção por SARS-CoV-2: relato de caso

2021 ◽  
Author(s):  
Sarah de Souza Almeida ◽  
Julianne Carvalho Dias Gaudio ◽  
Mônica Gomes de Almeida ◽  
Cristiane Alves de Oliveira
Keyword(s):  
Diabetes Mellitus ◽  
Rt Pcr ◽  
Terapia Intensiva

Introdução: Evidências da doença COVID-19 durante a gestação têm indicado maior risco de hospitalização, necessidade de ventilação mecânica e internação em unidade de terapia intensiva (UTI). Além disso, a COVID-19 parece estar associada a maior risco de pré-eclâmpsia, parto pré- termo e outros desfechos adversos. O melhor momento e via do parto na vigência da doença são questões ainda não completamente elucidadas com as evidências científicas disponíveis. Este artigo traz o relato de uma gestante com infecção por SARS-CoV-2 no terceiro trimestre. Relato de caso: Primigesta de 27 anos, com 37 semanas e 5 dias de idade gestacional, foi internada em enfermaria obstétrica para controle de diabetes mellitus gestacional (DMG). Na admissão, relatou história de quadro febril (37,8°C) há três dias e mialgia. O rastreio de SARS-CoV-2 com reação em cadeia da polimerase em tempo real (RT-PCR) em swab nasofaríngeo foi positivo. Exames admissionais demonstraram elevação de enzimas hepáticas e relação proteína/creatinina em amostra urinária alterada. Apesar da curva pressórica normal durante toda internação, foi aventada a possibilidade de pré-eclâmpsia sobreposta à infecção. A paciente manteve sintomas leves de COVID-19 até o oitavo dia de doença (febre baixa sem sintomas respiratórios). Em decisão conjunta com o serviço de Infectologia, foi indicada a interrupção da gestação por via vaginal, com 38 semanas e 3 dias, por DMG e piora dos parâmetros laboratoriais. Iniciou-se a indução medicamentosa do trabalho de parto com misoprostol. Após 18 horas de indução, a paciente evoluiu com taquipneia, leve redução da saturação de O2 e cardiotocografia com possível alteração do bem-estar fetal, e foi indicada a cesariana. O parto ocorreu sem intercorrências, e o recém-nascido apresentou boa vitalidade. No quarto dia pós-parto, a paciente evoluiu com pneumonia, deterioração clínica grave, necessidade de internação em UTI e ventilação mecânica por 15 dias. Após boa evolução, recebeu alta hospitalar em 29 dias da admissão. Conclusão: Evidências atuais demonstram que a COVID-19 na gravidez está associada ao aumento de morbidade materna grave e mortalidade, além de complicações neonatais. O momento ideal para a interrupção da gestação na COVID-19 ainda está em discussão. Alguns princípios podem ser usados para a tomada de decisão, como: indicação obstétrica, COVID-19 sem melhora apesar do tratamento instituído ou doença materna severa em que a antecipação do parto pareça possibilitar melhor manejo materno. A progressão da doença, a idade gestacional e o bem-estar fetal são os principais parâmetros a serem avaliados, contudo a segurança materna é a prioridade. A via de parto deve ser individualizada e considerar indicações obstétricas. O parto vaginal parece estar associado a melhor evolução clínica. A cesariana parece estar relacionada ao aumento de risco de deterioração clínica e de internação em UTI em grávidas com COVID-19. O estresse fisiológico induzido pela cirurgia parece ser o fator associado a essa piora.

scholarly journals Feeding Stimulates Sphingosine-1-Phosphate Mobilization in Mouse Hypothalamus

2019 ◽  
Vol 20 (16) ◽  
pp. 4008
Author(s):  
Valentina Vozella ◽  
Natalia Realini ◽  
Alessandra Misto ◽  
Daniele Piomelli
Keyword(s):  
De Novo ◽  
Sphingolipid Metabolism ◽  
Rt Pcr ◽  
Liquid Chromatography Tandem Mass ◽  
Sphingosine 1 Phosphate ◽  
Phosphate Mobilization ◽  
S1p Receptor ◽  
Free Feeding ◽  
G Protein Coupled ◽  
Sphingolipid Biosynthesis

Previous studies have shown that the sphingolipid-derived mediator sphingosine-1-phosphate (S1P) reduces food intake by activating G protein-coupled S1P receptor-1 (S1PR1) in the hypothalamus. Here, we examined whether feeding regulates hypothalamic mobilization of S1P and other sphingolipid-derived messengers. We prepared lipid extracts from the hypothalamus of C57Bl6/J male mice subjected to one of four conditions: free feeding, 12 h fasting, and 1 h or 6 h refeeding. Liquid chromatography/tandem mass spectrometry was used to quantify various sphingolipid species, including sphinganine (SA), sphingosine (SO), and their bioactive derivatives SA-1-phosphate (SA1P) and S1P. In parallel experiments, transcription of S1PR1 (encoded in mice by the S1pr1 gene) and of key genes of sphingolipid metabolism (Sptlc2, Lass1, Sphk1, Sphk2) was measured by RT-PCR. Feeding increased levels of S1P (in pmol-mg−1 of wet tissue) and SA1P. This response was accompanied by parallel changes in SA and dihydroceramide (d18:0/18:0), and was partially (SA1P) or completely (S1P) reversed by fasting. No such effects were observed with other sphingolipid species targeted by our analysis. Feeding also increased transcription of Sptlc2, Lass1, Sphk2, and S1pr1. Feeding stimulates mobilization of endogenous S1PR1 agonists S1P and SA1P in mouse hypothalamus, via a mechanism that involves transcriptional up-regulation of de novo sphingolipid biosynthesis. The results support a role for sphingolipid-mediated signaling in the central control of energy balance.

scholarly journals Regulation and Mechanism of miR-518d through the PPARα-Mediated NF-κB Pathway in the Development of Gestational Diabetes Mellitus

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Hui Qiu ◽  
Xuemin Liu ◽  
Shenshen Yao ◽  
Jiaren Zhou ◽  
Xue Zhang ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Gestational Diabetes ◽  
Gestational Diabetes Mellitus ◽  
Pregnant Women ◽  
Western Blotting ◽  
Transport Process ◽  
Inflammatory Factors ◽  
Rt Pcr ◽  
Expression Levels ◽  
Antagonist Group

Objectives. To observe the role of miR-518d in pregnant women with gestational diabetes mellitus (GDM) and its adjusting effects on PPARα and to explore the regulatory mechanisms of the NF-κB pathway in the development and progression of GDM. Methods. Placenta tissues and peripheral plasma were obtained from pregnant women with normal pregnancy and GDM, respectively, followed by the detections of miR-518d contents by RT-PCR and the expression levels of inflammatory factors using ELISA. Human placenta trophoblast cells (HTR8/SVneo) were cultured under the conditions of physiological glucose (PG group) and high glucose level (HG group). Cells in the HG group were transfected with miR-518d control, mimics, and inhibitors and were separately administered with a PPARα-specific antagonist (GW6471) and PPARα siRNA, and cells were divided into the following groups: HG+miR-518d control group (HGNC group), HG+miR-518d mimic group (HGM group), HG+miR-518d inhibitor group (HGI group), HGI+PPARα antagonist group, and HGI+PPARα siRNA group. The relative expression levels of miR-518d, PPARα, and its downstream genes and NF-κB signalling pathway-related genes were detected by RT-PCR and Western blotting. The contents of inflammatory factors were examined by Western blotting. A dual-luciferase report assay was performed to validate the correlations between miR-518d and PPARα. In this study, mouse GDM models were established to further prove the previous hypothesis with an in vivo experiment. A total of 40 C57BL/6J mice were randomly divided into the following groups: normal diet group (ControlMs), GDM group (GDMMs group), GDM+miR-518d antagomir group, and GDM+miR-518d antagomir+PPARα antagonist group. The mouse model of GDM was established by feeding with combined high-sugar and high-saturated fat diet and injecting streptozotocin (STZ) after 15-day feeding. Female and male mice were cocaged in the number ratio of 2 : 1, and the evidence of vaginal suppository detected in female mice was marked as D0 of pregnancy. The contents of total cholesterol (CH), triglyceride (TG), fast glucose, and insulin (INS) were examined using ELISA, followed by the evaluation of insulin resistance (IR). The related expression levels were also detected with the above methods shown in the previous cell culture. Results. miR-518d has a high expression level in placentas with GDM. As the target gene of miR-518d, PPARα was downregulated with the increased levels of miR-518d. When GDM occurs, inflammatory responses were elevated, stimulating the nuclear transport process of NF-κB. Activated NF-κB triggered the phosphorylation of IKKβ and IκBα. Conclusions. High expression of miR-518d was observed in the development of GDM. In this study, we validated that miR-518d negatively regulates the expression of PPARα and triggers the nuclear transport process of NF-κB and phosphorylation of pathway-associated proteins leading to an inflammatory response and the development of GDM.

scholarly journals MicroRNA-133a impairs perfusion recovery after hindlimb ischemia in diabetic mice

2018 ◽  
Vol 38 (4) ◽  
Author(s):  
Lingdan Chen ◽  
Chunli Liu ◽  
Dejun Sun ◽  
Tao Wang ◽  
Li Zhao ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Endothelial Cells ◽  
Arterial Disease ◽  
Tube Formation ◽  
Cgmp Level ◽  
Diabetic Mice ◽  
Rt Pcr ◽  
Patients With Diabetes ◽  
Peripheral Arterial ◽  
Ischemic Muscle

Objective: Peripheral arterial disease (PAD) patients with diabetes mellitus suffer from impaired neovascularization after ischemia which results in poorer outcomes. MicroRNA (miR)-133a is excessively expressed in endothelial cells under diabetic conditions. Here, we test whether diabetes-induced miR-133a up-regulation is involved in the impaired capability of neovascularization in experimental PAD models. Methods and results: MiR-133a level was measured by quantitative RT-PCR and showed a higher expression level in the ischemic muscle from diabetic mice when compared with nondiabetic mice. Knockdown of miR-133a using antagomir improved perfusion recovery and angiogenesis in experimental PAD model with diabetes day 21 after HLI. On the other hand, overexpression of miR-133a impaired perfusion recovery. Ischemic muscle was harvested day 7 after experimental PAD for biochemical test, miR-133a antagonism resulted in reduced malondialdehyde, and it increased GTP cyclohydrolase 1 (GCH1), and cyclic guanine monophosphate (cGMP) levels. In cultured endothelial cells, miR-133a antagonism resulted in reduced reactive oxygen species level, and it increased tube formation, nitric oxide (NO), and cGMP level. Moreover, miR-133a antagonism-induced angiogenesis was abolished by GCH1 inhibitor. In contrary, miR-133a overexpression impairs angiogenesis and it reduces GCH1, NO, and cGMP levels in nondiabetic models. Conclusion: Diabetes mellitus-induced miR-133a up-regulation impairs angiogenesis in PAD by reducing NO synthesis in endothelial cells. MiR-133a antagonism improves postischemic angiogenesis.

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