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1.
Arq. gastroenterol ; 59(2): 238-243, Apr.-June 2022. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1383855

ABSTRACT

ABSTRACT Background: Inflammatory bowel disease (IBD) comprises the spectrum between Crohn's disease (CD) and ulcerative colitis (UC), a condition whose prevalence in countries such as Brazil has increased significantly in recent years. Changes in the intestinal epithelial barrier function and, consequently, an increase in intestinal permeability, have been suggested as important factors in the pathogenesis of different autoimmune conditions, including IBD. Therefore, there is a need for a practical tool to assess gut barrier integrity in these patients. Objective: To study factors associated with serum zonulin levels, a marker of intestinal permeability, in patients with IBD. Methods: This was a cross-sectional observational study that included 117 patients with IBD and 32 healthy controls. Disease activity was assessed by the Simple Clinical Colitis Activity Index (SCCAI) in UC and by the Harvey-Bradshaw Index (HBI) in CD subjects. Zonulin levels were measured by ELISA and inflammatory cytokines by Cytometric Bead Array, using commercially available kits. Results: The mean age of IBD patients was 44.0±15.9 years, 66.7% were female, 57 subjects were diagnosed with CD and 60 with UC. At evaluation, clinical remission was observed in 56.7% of CD patients and in 59.2% of UC subjects. No differences were observed in zonulin levels when comparing IBD patients with the control group (95.28 ng/mL vs 96.61 ng/mL, P=0.573) and when comparing patients with CD to those with UC (79.68 ng/mL vs 106.10 ng/mL, P=0.887). Among IBD group, zonulin concentrations were higher among females, correlated positively with body mass index (BMI) and age; and negatively with hemoglobin and hematocrit. In patients with UC, zonulin correlated negatively with hemoglobin, hematocrit, and albumin; and positively with BMI and SCCAI. Among CD patients, zonulin was positively correlated with age and BMI, but not with HBI. No correlations were observed between zonulin and circulating cytokines in IBD patients. Conclusion: In this cohort mostly comprised of patients in clinical remission, serum zonulin levels were not higher in patients with IBD than healthy controls, and correlated with variables not linked to baseline disease, such as sex, age and BMI. However, zonulin correlated with clinical and laboratory parameters of disease severity and activity among subjects with UC, but not among patients with CD. These findings indicate a potential role for zonulin as a biomarker in IBD, particularly in UC.


RESUMO Contexto: A doença inflamatória intestinal (DII) compreende o espectro entre a doença de Crohn (DC) e a colite ulcerativa, condição esta cuja prevalência em países como o Brasil vem aumentando significativamente nos últimos anos. Alterações na função da barreira epitelial intestinal e, consequentemente, um aumento da permeabilidade intestinal, têm sido sugeridos como fatores importantes envolvidos na patogênese de diferentes condições autoimunes, dentre elas, a DII. Desta forma, existe a necessidade de uma ferramenta prática para avaliar a integridade da barreira epitelial intestinal nestes pacientes. Objetivo: Estudar os fatores associados com os níveis séricos de zonulina, um marcador da permeabilidade intestinal, em pacientes com DII. Métodos: Estudo observacional transversal que incluiu 117 pacientes com DII e 32 indivíduos que compuseram o grupo controle. A atividade da doença foi avaliada pelo Simple Cliniical Colitis Activity Index (SCCAI) na colite ulcerativa e pelo índice de Harvey-Bradshaw (IHB) em pacientes com DC. Os níveis de zonulina foram quantificados por ELISA e os níveis das citocinas inflamatórias pelo Cytometric Bead Array, utilizando kits comercialmente disponíveis. Resultados: A média de idade dos pacientes com DII foi de 44,0±15,9 anos, 66,7% eram do sexo feminino, 57 pacientes eram portadores de DC e 60 pacientes eram portadores de colite ulcerativa. No momento da avaliação clínico-laboratorial, 56,7% dos pacientes com DC encontravam-se em remissão clínica e, dentre os pacientes com colite ulcerativa, 59,2% deles assim se encontravam. Não foram observadas diferenças nos níveis séricos de zonulina entre pacientes com DII e grupo controle (95,28 ng/mL vs 96,61 ng/mL; P=0,573), assim como entre pacientes com DC e pacientes com colite ulcerativa (79,68 ng/mL vs 106,10 ng/mL, P=0,887). Dentre os pacientes com DII, as concentrações de zonulina foram mais elevadas no sexo feminino e correlacionaram-se positivamente com o índice de massa corporal (IMC) e com a idade, correlacionando-se negativamente com os níveis de hemoglobina e hematócrito. Nos pacientes com colite ulcerativa, as concentrações de zonulina correlacionaram-se negativamente com os parâmetros hemoglobina, hematócrito e albumina e, positivamente, com o IMC e com o SCCAI. Dentre os pacientes com DC, a zonulina sérica correlacionou-se positivamente com a idade e com o IMC, mas não com o IHB. Não foram observadas correlações entre os níveis de zonulina e as citocinas circulantes nos pacientes com DII. Conclusão: Nesta coorte constituída majoritariamente por pacientes em remissão clínica, os níveis séricos de zonulina não se mostraram aumentados em pacientes com DII em relação a indivíduos controles e correlacionaram-se com variáveis não relacionadas à doença de base, como com o sexo, com a idade e com o IMC. No entanto, os níveis séricos de zonulina correlacionaram-se com parâmetros clínicos e laboratoriais de gravidade e atividade da doença dentre os pacientes com colite ulcerativa, mas não dentre os pacientes com DC. Estes achados indicam um potencial papel da zonulina sérica como um biomarcador na DII, principalmente na colite ulcerativa.

2.
Braz. dent. j ; 33(2): 1-11, Mar.-Apr. 2022. tab, graf
Article in English | LILACS-Express | LILACS, BBO | ID: biblio-1374628

ABSTRACT

Abstract The aim was to assess the physicochemical properties and the penetration into dentinal tubules of calcium hypochlorite solution [Ca(OCl)2], with or without surfactants. The surfactants benzalkonium chloride, cetrimide, Tween 80 and Triton X-100 were mixed at different concentrations with sodium hypochlorite solution (NaOCl), Ca(OCl)2 and distilled water (control). Once the critical micellar concentration (CMC) of the surfactants in Ca(OCl)2 and NaOCl was determined, pH, free chlorine, surface tension and free calcium ions were evaluated. The penetration into dentinal tubules of NaOCl and Ca(OCl)2, with or without benzalkonium chloride and Triton X-100 [surfactants that promoted the lowest surface tension of Ca(OCl)2], was assessed using human premolars stained with crystal violet. The statistical tests were one-way ANOVA and Tukey's post-test, Kruskal-Wallis and Dunn's post-test, two-way ANOVA and Bonferroni's post-test, and t-test; depending on the assay. The addition of surfactants reduced the surface tension of NaOCl and Ca(OCl)2, and did not alter the pH or the free available chlorine of either solution. The addition of all surfactants increased the availability of free calcium ions in Ca(OCl)2, especially benzalkonium chloride. Ca(OCl)2 exhibited lower penetration into dentinal tubules than NaOCl, and the addition of surfactants did not improve the penetration of Ca(OCl)2, but did increase the penetration of NaOCl. It can be concluded that the addition of surfactants to Ca(OCl)2 did not increase the penetration into dentinal tubules, but it did promote lower surface tension, without changing the pH or free available chlorine values, and higher availability of free calcium ions in Ca(OCl)2.


Resumo O objetivo foi avaliar as propriedades físico-químicas e a penetrabilidade nos túbulos dentinários da solução de hipoclorito de cálcio [Ca(OCl)2], com ou sem surfactantes. Os surfactantes cloreto de benzalcônio, cetrimida, Tween 80 e Triton X-100 foram misturados em diferentes concentrações com a solução de hipoclorito de sódio (NaOCl), Ca(OCl)2 e água destilada (controle). Uma vez determinada a concentração micelar crítica (CMC) dos surfactantes em Ca(OCl)2 e NaOCl, foram avaliados o pH, cloro livre, tensão superficial e íons de cálcio livre. A penetrabilidade nos túbulos dentinários de NaOCl e Ca(OCl)2, com ou sem cloreto de benzalcônio e Triton X-100 [surfactantes que promoveram a menor tensão superficial de [Ca(OCl)2] foi avaliada utilizando pré-molares humanos corados com cristal violeta. Os testes estatísticos foram ANOVA de uma via e pós-teste de Tukey, Kruskal-Wallis e pós-teste de Dunn, ANOVA de duas vias e pós-teste de Bonferroni, e teste t; dependendo do ensaio. A adição de surfactantes reduziu a tensão superficial do NaOCl e Ca(OCl)2, e não alterou o pH ou cloro livre das soluções. A adição de todos os surfactantes aumentou a disponibilidade de íons de cálcio livre de Ca(OCl)2, principalmente o cloreto de benzalcônio. Ca(OCl)2 apresentou menor penetrabilidade nos túbulos dentinários do que NaOCl, e a adição de surfactantes não aumentou a penetrabilidade de Ca(OCl)2, mas aumentou a penetrabilidade de NaOCl. Pode-se concluir que a adição de surfactantes no Ca(OCl)2 não aumentou a penetrabilidade nos túbulos dentinários, mas promoveu menor tensão superficial, sem alterar os valores de pH ou cloro livre, e maior disponibilidade de íons de cálcio livre em Ca(OCl)2.

3.
Article in Chinese | WPRIM | ID: wpr-920798

ABSTRACT

Objective To establish bovine corneal opacity and permeability (BCOP) test, and determine its predictive ability for the eye irritation evaluation of cosmetics. Methods A total of ten reference chemicals were selected to establish the BCOP test. Then eye irritation of 16 routinely collected cosmetics in our laboratory was predicted. In vitro scores were calculated by the change in the opacity and sodium fluorescein permeability after exposure to the testing cosmetics, and subsequently compared with the historical data by Draize test. Results Reference chemicals with known irritation classification were correctly classified by the BCOP test, which was consistent with the classification of UN globally harmonized system of classification and labeling of chemicals. Moreover, the specificity of the BCOP test for the classification of non-irritating cosmetics samples was 80.0% (8/10), and the sensitivity for weak to mild irritating cosmetics samples was 83.3% (5/6). The BCOP test demonstrated an overall classification consistency of 81.3% (13/16) with in vivo test. Conclusion BCOP test may be independently used to identify chemicals with potential eye irritation and serious eye damage, suggesting it is significant for in vitro integrated test strategy for predicting eye irritation due to cosmetics.

4.
Article in Chinese | WPRIM | ID: wpr-933470

ABSTRACT

Objective:To investigate the effect of interleukin-33 (IL-33) on lipopolysaccharide (LPS)-induced permeability of rat cardiac microvascular endothelial cells (RCMECs).Methods:RCMECs were cultured in vitro to be divided into control group, LPS group, IL-33 group and LPS+IL-33 group. The effect of IL-33 on the proliferation of RCMECs was detected by cell counting reagent (CCK8). Fluorescein isothiocyanate (FITC)-dextran assay was used to evaluate the permeability of RCMECs. The expression of vascular endothelial calmodulin, ras homologous gene family (Rho) member A (RhoA) and phosphorylated Rho-associated coiled-coil-containing protein kinase (p-ROCK2) proteins were tested by western blot. High-throughput sequencing and gene ontology (GO) were performed for gene expression in LPS and LPS+IL-33 groups.Results:No significant effect of IL-33 at 10-50 ng/ml on the proliferation of RCMECs was observed ( P>0.05). Compared with the control group, the permeability of RCMECs (permeability coefficient ratio 1.404±0.029 vs. 1.000±0.200, P<0.05) was significantly increased in LPS group and the expression of vascular endothelial calmodulin (relative gray value 0.429 5±0.012 9 vs. 0.594 9±0.014 2, P<0.05) was down-regulated, while the permeability of monolayers (permeability coefficient ratio, 0.948±0.013, P<0.01) was decreased in LPS+IL-33 group and the expression of vascular endothelial calmodulin (relative grayscale value 0.549 1±0.012 0, P<0.005) was up-regulated compared with the LPS group. High-throughput sequencing data revealed that the differential genes downregulated in the LPS and LPS+IL-33 groups were associated with cytoskeleton and Rho signaling pathway. Compared with the control group, RhoA (relative gray value 0.211 4±0.009 9 vs. 0.135 0±0.007 6, P<0.000 1) and p-ROCK (relative gray value 0.656 3±0.013 2 vs. 0.503 6±0.036 2, P<0.000 1) protein expression was upregulated in the LPS group. When compared with LPS group, RhoA (relative gray value 0.157 7±0.010 7, P=0.000 2), p-ROCK (relative gray value 0.427 7±0.003 8, P<0.000 1) protein expression was decreased in LPS+IL-33 group. Conclusion:IL-33 may improve LPS-induced hyperpermeability of RCMECs by inhibiting RhoA and p-ROCK protein expression in Rho/Rho-associated coiled-coil-containing protein kinase signaling pathway.

5.
Article in Chinese | WPRIM | ID: wpr-932579

ABSTRACT

Objective:To explore the mechanism of lysosomal membrane permeabilization(LMP)inuranyl acetate-induced death of human kidney proximal tubular epithelial HK-2 cells.Methods:HK-2 cells were exposed to uranyl acetate at concentrations of 100, 300 and 600 μmol/L for 24 h, then in tracellular reactive oxygen species (ROS)and mitochondrial superoxide were measured by DCFH-DA and MitoSOX probe, respectively. HK-2 cells were divided into four groups: blank control group, NAC or CA-074 Me group, uranyl acetate exposure group and uranyl acetate exposure plus NAC or CA-074 Me group. Two-color immune of luorescence staining was used to detect the co-localization of galectin-1 and lysosomal associated membrane protein-1 (LAMP-1) to measure the extent of LMP, and to detect the non- co-localization of cathepsin B and LAMP-1 to reflect the release of cathepsin B in lysosomes. Calcein-AM/PI double staining method was used to detect cell death. One-color immune of luorescence staining of cleaved-caspase-3 expression was used to detect apoptosis. Results:Intracellular ROS and mitochondrial superoxide levels were significantly increased in HK-2 cells after exposure with 100, 300 and 600 μmol/L uranyl acetate for 24 h, about 1.1-2.5 times or 4.0-28 times, respectively( tROS=17.98, 11.84, 11.75, P< 0.05; tmitochondrial superoxide=6.14, 16.02, 13.06, P< 0.05), and they also increased with uranyl acetate concentrations ( tROS=10.10, 10.37, 5.59, P< 0.05; tmitochondrial superoxide=21.50, 15.16, 5.93, P< 0.05). The percentage of co-localization of galectin-1 and LAMP-1 and the percentage of non- co-localization of cathepsin B and LAMP-1 were markedly increased in HK-2 cells after exposure with 600 μmol/L uranyl acetate for 24 h, 5.4-6.7 times or 1.5-2.1 times, respectively ( tGalectin-1=15.85, 12.70, P< 0.05; tCathepsin B=5.95, 6.69, P< 0.05), but these increases were inhibited by NAC ( tGalectin-1=4.74, P<0.05; tCathepsin B=4.51, P< 0.05). Moreover, the cell death rate and the cleaved-caspase-3 expression level were also significantly increased in HK-2 cells after exposure with 600 μmol/L uranyl acetate for 24 h, about 28-47 times or 2.4-6.0 times, respectively( tPI=30.40, 10.34, P<0.05; tCleaved-caspase-3=18.49, 9.52, P<0.05), and these increases were obviously diminished by CA-074 Me ( tPI= 6.76, P<0.05; tCleaved-caspase-3=13.47, P<0.05). Conclusions:Exposure to uranyl acetate induces a burst of intracellular ROSthat leads to LMP and consequently causes leakage of cathepsin B from lysosomes to cytoplasm, in turn triggering the lysosomal-dependent cell death and mitochondrial-regulated apoptosis of HK-2 cells.

6.
Acta Pharmaceutica Sinica B ; (6): 92-106, 2022.
Article in English | WPRIM | ID: wpr-929283

ABSTRACT

Nanoparticulate drug delivery systems (Nano-DDSs) have emerged as possible solution to the obstacles of anticancer drug delivery. However, the clinical outcomes and translation are restricted by several drawbacks, such as low drug loading, premature drug leakage and carrier-related toxicity. Recently, pure drug nano-assemblies (PDNAs), fabricated by the self-assembly or co-assembly of pure drug molecules, have attracted considerable attention. Their facile and reproducible preparation technique helps to remove the bottleneck of nanomedicines including quality control, scale-up production and clinical translation. Acting as both carriers and cargos, the carrier-free PDNAs have an ultra-high or even 100% drug loading. In addition, combination therapies based on PDNAs could possibly address the most intractable problems in cancer treatment, such as tumor metastasis and drug resistance. In the present review, the latest development of PDNAs for cancer treatment is overviewed. First, PDNAs are classified according to the composition of drug molecules, and the assembly mechanisms are discussed. Furthermore, the co-delivery of PDNAs for combination therapies is summarized, with special focus on the improvement of therapeutic outcomes. Finally, future prospects and challenges of PDNAs for efficient cancer therapy are spotlighted.

7.
Article in English | WPRIM | ID: wpr-929260

ABSTRACT

Wuzi-Yanzong-Wan (WZYZW) is a classic prescription for male infertility. Our previous investigation has demonstrated that it can inhibit sperm apoptosis via affecting mitochondria, but the underlying mechanisms are unclear. The purpose of the present study was to explore the actions of WZYZW on mitochondrial permeability transition pore (mPTP) in mouse spermatocyte cell line (GC-2 cells) opened by atractyloside (ATR). At first, WZYZW-medicated serum was prepared from rats following oral administration of WZYZW for 7 days. GC-2 cells were divided into control group, model group, positive group, as well as 5%, 10%, 15% WZYZW-medicated serum group. Cyclosporine A (CsA) was used as a positive control. 50 μmol·L-1 ATR was added after drugs incubation. Cell viability was assessed using CCK-8. Apoptosis was detected using flow cytometry and TUNEL method. The opening of mPTP and mitochondrial membrane potential (MMP) were detected by Calcein AM and JC-1 fluorescent probe respectively. The mRNA and protein levels of voltage-dependent anion channel 1 (VDAC1), cyclophilin D (CypD), adenine nucleotide translocator (ANT), cytochrome C (Cyt C), caspase 3, 9 were detected by RT-PCR (real time quantity PCR) and Western blotting respectively. The results demonstrated that mPTP of GC-2 cells was opened after 24 hours of ATR treatment, resulting in decreased MMP and increased apoptosis. Pre-protection with WZYZ-medicated serum and CsA inhibited the opening of mPTP of GC-2 cells induced by ATR associated with increased MMP and decreased apoptosis. Moreover, the results of RT-qPCR and WB suggested that WZYZW-medicated serum could significantly reduce the mRNA and protein levels of VDAC1 and CypD, Caspase-3, 9 and CytC, as well as a increased ratio of Bcl/Bax. However, ANT was not significantly affected. Therefore, these findings indicated that WZYZW inhibited mitochondrial mediated apoptosis by attenuating the opening of mPTP in GC-2 cells. WZYZW-medicated serum inhibited the expressions of VDAC1 and CypD and increased the expression of Bcl-2, which affected the opening of mPTP and exerted protective and anti-apoptotic effects on GC-2 cell induced by ATR.


Subject(s)
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine , Animals , Atractyloside/pharmacology , Cyclophilin D , Male , Matrix Metalloproteinases , Mice , Mitochondrial Membrane Transport Proteins/metabolism , Mitochondrial Permeability Transition Pore , RNA, Messenger , Rats
8.
Article in Chinese | WPRIM | ID: wpr-940186

ABSTRACT

ObjectiveTo compare the differences in resistance and structure of skin between acupoints and non-acupoints, and to study the difference in skin permeability characteristics of Corydalis Rhizoma total alkaloid patches (CTTP) after administration at Shenque acupoint and non-acupoint, so as to provide experimental support for its clinical acupoint application to prevent and treat chronic pain. MethodTaking corydaline (CD), tetrahydropalmatine (THP) and corydalis L (CDL) as evaluation indexes, and the quantitative analysis was carried out by high performance liquid chromatography (HPLC). The mobile phase was methanol-0.04 mol·L-1 phosphoric acid aqueous solution (70∶30, pH 6.0 adjusted with triethylamine), the detection wavelength was 281 nm. In vitro transdermal test in Franz diffusion cell and in vivo transdermal test were used to study the skin permeability characteristics of CTTP through Shenque acupoint and non-acupoint administration. At the same time, the skin resistance between Shenque acupoint and non-acupoint was measured before and after the administration, and the distribution of the drug in each layer of the skin was compared by freezing sectioning, and visual verification was performed with fluorescence inverted microscope. ResultAfter 24 h of administration, the results of in vivo and in vitro experiments showed that the cumulative permeation and retention of CD, THP and CDL at Shenque acupoint skin were higher than those at non-acupoint skin (P<0.05, P<0.01), the skin resistance of Shenque acupoint was lower than that of non-acupoint at all time points. The fluorescence microscopic observation results showed that the drug content of each layer of the skin was all Shenque acupoint>non-acupoint, indicating that the skin of Shenque acupoint had better effect on drug penetration and storage than non-acupoint. ConclusionThe 24 h cumulative permeation and retention of CTTP in Shenque acupoint skin are higher than those in non-acupoint skin, and the mechanism may be related to the thin skin, low electrical resistance and large number of hair follicle bodies at Shenque acupoint.

9.
Article in Chinese | WPRIM | ID: wpr-940154

ABSTRACT

ObjectiveTo compare the differences in resistance and structure of skin between acupoints and non-acupoints, and to study the difference in skin permeability characteristics of Corydalis Rhizoma total alkaloid patches (CTTP) after administration at Shenque acupoint and non-acupoint, so as to provide experimental support for its clinical acupoint application to prevent and treat chronic pain. MethodTaking corydaline (CD), tetrahydropalmatine (THP) and corydalis L (CDL) as evaluation indexes, and the quantitative analysis was carried out by high performance liquid chromatography (HPLC). The mobile phase was methanol-0.04 mol·L-1 phosphoric acid aqueous solution (70∶30, pH 6.0 adjusted with triethylamine), the detection wavelength was 281 nm. In vitro transdermal test in Franz diffusion cell and in vivo transdermal test were used to study the skin permeability characteristics of CTTP through Shenque acupoint and non-acupoint administration. At the same time, the skin resistance between Shenque acupoint and non-acupoint was measured before and after the administration, and the distribution of the drug in each layer of the skin was compared by freezing sectioning, and visual verification was performed with fluorescence inverted microscope. ResultAfter 24 h of administration, the results of in vivo and in vitro experiments showed that the cumulative permeation and retention of CD, THP and CDL at Shenque acupoint skin were higher than those at non-acupoint skin (P<0.05, P<0.01), the skin resistance of Shenque acupoint was lower than that of non-acupoint at all time points. The fluorescence microscopic observation results showed that the drug content of each layer of the skin was all Shenque acupoint>non-acupoint, indicating that the skin of Shenque acupoint had better effect on drug penetration and storage than non-acupoint. ConclusionThe 24 h cumulative permeation and retention of CTTP in Shenque acupoint skin are higher than those in non-acupoint skin, and the mechanism may be related to the thin skin, low electrical resistance and large number of hair follicle bodies at Shenque acupoint.

10.
Article in Chinese | WPRIM | ID: wpr-928181

ABSTRACT

This study aims to establish a method for determination of paeonol(Pae), eugenol(Eug), and piperine(Pip) content in receptor liquid and research on the permeability and pharmacokinetics of Huoxue Zhitong gel patch and microemulsion gel. The Franz diffusion experiment was conducted to assess the percutaneous permeability, and the microdialysis method was employed to assess pharmacokinetics of Huoxue Zhitong gel patch and microemulsion gel. The content of Pae, Eug, and Pip in receptor liquid in vitro and in vivo was determined by HPLC and UPLC-MS. The Q_n and J_(ss) of Pae, Eug, and Pip in the gel patch were significantly higher than those in the microemulsion gel, indicating that the drug release was faster in the gel patch. The C_(max), AUC_(0-760), and MRT of Pae, Eug, and Pip in the gel patch were higher than those in the microemulsion gel, indicating that the gel patch can promote the penetration and prolong the skin residence of the drug. The results of this study provide reference for improving the dosage form of Huoxue Zhitong patch.


Subject(s)
Administration, Cutaneous , Chromatography, Liquid , Emulsions , Permeability , Skin/metabolism , Skin Absorption , Tandem Mass Spectrometry
11.
Article in Chinese | WPRIM | ID: wpr-928020

ABSTRACT

The present study investigated the effect of active components of Descurainia sophia on allergic asthma and explored the underlying mechanism. SD male rats were randomly divided into a normal group(NC), a model group(M), a D. sophia decoction group(DS), a D. sophia fatty oil group(FO), a D. sophia flavonoid glycoside group(FG), a D. sophia oligosaccharide group(Oli), and a positive drug dexamethasone group(Y). The allergic asthma model was induced in rats by intraperitoneal injection of ovalbumin(OVA) and aluminum hydroxide gel adjuvant(sensitization) and atomization of OVA solution(excitation). After modeling, asthma-related indicators, tracheal phenol red excretion, inflammatory cell levels in the peripheral blood, lung permeability index(LPI), and oxygenation index(OI) of rats were detected. The pathological changes of lung tissues were observed by HE staining. Enzyme-linked immunosorbent assay(ELISA) was used to detect the content of inflammatory factors immunoglobulin E(IgE), interleukin-4(IL-4), and interferon-γ(IFN-γ) in the bronchoalveolar lavage fluid(BALF) and the content of endothelin-1(ET-1) and angiotensin-converting enzyme(ACE) in lung tissue homogenate. The serum content of nitric oxide(NO) was detected by colorimetry. Western blot was employed to determine the protein expression of Toll-like receptor 4(TLR4), nuclear factor κB-p65(NF-κB-p65), phosphorylated NF-κB-p65(p-NF-κB-p65), myosin light chain kinase(MLCK), vascular endothelial cadherin(VE cadherin), connexin 43, and claudin 5, and the mechanism of active components of D. sophia on allergic asthma was explored. As revealed by the results, the M group showed extensive infiltration of inflammatory cells around the bronchus of the lung tissues of the allergic asthma rats, thickened bronchial wall, severely deformed alveolar structure, increased number of wheezes, the content of IgE, IL-4, ET-1, and ACE, inflammatory cells, and LPI, and reduced latency of asthma, tracheal phenol red excretion, IFN-γ, NO content, and OI. After the intervention of the active components of D. sophia, the DS, FO, FG, Oli, and Y groups showed improved asthma-related indicators, tracheal phenol red excretion, and lung tissue lesions in allergic asthma rats, and the effects in the FO and Oli groups were superior. The content of inflammatory factors in BALF was recovered in the DS, FO, and Y groups and the FG and Oli groups. The number of inflammatory cells in rats was reduced in the DS and FO groups, and the FG, Oli, and Y groups to varying degrees, and the effect in the FO group was superior. DS, FO, Oli, and Y reduced ET-1, ACE, and LPI and increased NO and OI. FG recovered NO, ET-1, ACE, LPI, and OI to improve lung epithelial damage and permeability. Further investigation of inflammation-related TLR4/NF-κB pathways, MLCK, and related skeleton protein levels showed that TLR4, NF-κB-p65, p-NF-κB-p65, and MLCK levels were increased, and VE cadherin, connexin 43, and claudin 5 were reduced in the M group. DS, FO, FG, Oli, and Y could reduce the protein expression related to the TLR4 pathway to varying degrees, and regulate the protein expression of MLCK, VE cadherin, connexin 43, and claudin 5. It is inferred that the active components of D. sophia improve lung permeability in rats with allergic asthma presumedly by regulating the TLR4/NF-κB signaling pathway to improve airway inflammation, mediating MLCK and connexin, and regulating epithelial damage.


Subject(s)
Animals , Asthma/drug therapy , Bronchoalveolar Lavage Fluid , Inflammation/metabolism , Lung , Male , Permeability , Rats
12.
Acta Pharmaceutica Sinica ; (12): 409-418, 2022.
Article in Chinese | WPRIM | ID: wpr-922916

ABSTRACT

We investigated the ability of Dracocephalum moldavica (EPDM) flavonoids to protect human brain microvascular endothelial cells (HBMECs) from necroptosis induced by ischemia-reperfusion injury. To mimic the process of cerebral ischemia-reperfusion injury, a necroptosis model was established by treatment with the pan-cysteine aspartic acid protease (caspase) inhibitor Z-VAD-FMK combined with oxygen-glucose deprivation/re-oxygenation (OGD/R) injury using HBMECs. Cell proliferation and cytotoxicity (cell counting kit-8, CCK-8) was used to measure cell viability. A Hoechst33342/PI fluorescent double-staining method was exploited to determine the rate of cell necroptosis. A commercial kit was used to detect lactate dehydrogenase in the cell culture supernate. DCFH-DA probes, calcein AM and JC-1 probes were used to measure changes in ROS production, mitochondrial membrane permeability transformation pore (MPTP) opening and mitochondrial membrane potential (MMP), respectively. Enzyme-linked immunosorbent assay (ELISA) kits were chosen to detect the release of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6). Western blotting was used to detect necroptosis-related proteins. The results show that relative to control group, Z-VAD-FMK combined with OGD/R injury reduced cell viability, increased the necroptosis rate and the levels of LDH and ROS in HBMECs. The MPTP of the model group cells opened and the MMP reduced. TNF-α, IL-1β, and IL-6 levels were significantly elevated. Furthermore, the expression of receptor-interacting protein kinase 3 (RIP3) and mitochondrial phosphoglycerate mutase 5 (PGAM5) was significantly increased, accompanied by an increase of phosphorylated mixed-lineage kinase domain-like protein (p-MLKL)/MLKL. EPDM partially reversed the changes of the above-mentioned factors in HBMECs induced by Z-VAD-FMK plus OGD/R injury. These results indicate that EPDM may protect HBMECs from cerebral ischemia-reperfusion injury by inhibiting the RIP3/MLKL/PGAM5 pathway and MPTP opening to maintain mitochondrial function, thereby providing a scientific basis for the use of EPDM in the treatment of cerebral ischemia-related diseases.

13.
Acta Pharmaceutica Sinica ; (12): 802-808, 2022.
Article in Chinese | WPRIM | ID: wpr-922904

ABSTRACT

A blood-brain barrier microfluidic chip platform for studying the permeability of active components in traditional Chinese medicine was developed. This model used primary human brain microvascular endothelial cells on a microfluidic chip consisting of two perpendicularly-crossing channels and a single layer porous polycarbonate membrane. The physiological shear stress in the human vasculature was also modeled in this device. Cell viability on the chip was monitored by cell staining and immunofluorescence staining. The cells spread well and the structure of an intercellular adhesion protein was satisfactory. The permeability of fluorescent tracers and three model drugs and the functional expression of P-glycoprotein (P-gp)on the blood-brain barrier were investigated. The results show that the apparent permeability coefficients (Papp) of the fluorescent tracers and three model drugs were consistent with those reported in the literature, and P-gp on the chip showed normal function, indicating that there was a complete structure and a functional BBB. The permeability of six active components of traditional Chinese medicine was investigated through this microfluidic chip and the drug concentration was determined by HPLC-MS/MS to obtain the Papp of each component. The Papp of corydaline was (4.51 ± 1.90)×10-7 cm·s-1, the Papp of tetrahydropalmatine was (9.10 ± 6.59)×10-7 cm·s-1, and the Papp of imperatorin was (9.38 ± 2.53)×10-7 cm·s-1; the concentration of isoimperatorin, baicalin and chlorogenic acid was below the limit of quantification, which suggested that isoimperatorin, baicalin and chlorogenic acid have poor permeability in this BBB chip. This blood-brain barrier microfluidic platform possesses a complete barrier function and near-physiological conditions and could be a valuable in vitro tool for drug permeability evaluation.

14.
Braz. j. med. biol. res ; 55: e12268, 2022. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1403912

ABSTRACT

Disruption of pulmonary endothelial permeability and associated barrier integrity increase the severity of acute respiratory distress syndrome (ARDS). This study investigated the potential ability of the human immunodeficiency virus-1 (HIV-1) integrase inhibitor raltegravir to protect against acute lung injury (ALI) and the underlying mechanisms. Accordingly, the impact of raltegravir treatment on an in vitro lipopolysaccharide (LPS)-stimulated human pulmonary microvascular endothelial cell (HPMEC) model of ALI and an in vivo LPS-induced two-hit ALI rat model was examined. In the rat model system, raltegravir treatment alleviated ALI-associated histopathological changes, reduced microvascular permeability, decreased Evans blue dye extravasation, suppressed the expression of inflammatory proteins including HMGB1, TLR4, p-NF-κB, NLRP3, and MPO, and promoted the upregulation of protective proteins including claudin 18.1, VE-cadherin, and aquaporin 5 as measured via western blotting. Immunohistochemical staining further confirmed the ability of raltegravir treatment to reverse LPS-induced pulmonary changes in NLRP3, claudin 18.1, and aquaporin 5 expression. Furthermore, in vitro analyses of HPMECs reaffirmed the ability of raltegravir to attenuate LPS-induced declines in VE-cadherin and claudin 18.1 expression while simultaneously inhibiting NLRP3 activation and reducing the expression of HMGB1, TLR4, and NF-kB, thus decreasing overall vascular permeability. Overall, our findings suggested that raltegravir may represent a viable approach to treating experimental ALI that functions by maintaining pulmonary microvascular integrity.

15.
J. appl. oral sci ; 30: e20220082, 2022. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1386011

ABSTRACT

Abstract Objectives To analyze the effect of 5 toothpastes containing different percentages of S-PRG fillers compared to NaF toothpaste and NaF varnish on the dentin hydraulic conductance (Lp). Methodology Dentin disks (1.0±0.2 mm thickness) were cut from third molars, and their Lp values were evaluated using Flodec. The specimens were allocated into 7 groups (n=8). The minimum (smear layer) and the maximum (after acid etching) Lp values were recorded. Lp was also assessed after treatment with either a 0wt.%, 1wt.%, 5wt.%, 20wt.%, or 30wt.% S-PRG toothpaste, a NaF toothpaste, or a NaF varnish. Toothpastes were applied by brushing for 15 s, allowing it to settle for 1 min, and rinsing with deionized water. The NaF varnish was applied for 4 min and was removed with a probe. Specimens were exposed to citric acid (6%, pH 2.1, 1 min) and their final Lp was recorded. The pH of all products was recorded (n=3) and specimens from each group were analyzed by Laser Scanning Confocal Microscopy (LSCM). Data were subjected to 2-way repeated measures ANOVA and post-hoc Bonferroni (a=0.05). Results The highest Lp reduction was noticed for the 5wt.% S-PRG toothpaste, NaF toothpaste, and NaF varnish. However, the toothpastes containing 5wt.%, 20wt.%, and 30wt.% of S-PRG were similar to all toothpastes but differed from the NaF varnish. After erosion, all groups retrieved their maximum Lp values, except for the NaF varnish. The LSCM evidenced deposits on the surface of specimens treated with 5%, 20%, and 30% S-PRG-based toothpastes and NaF toothpaste. Even more deposits were observed for the NaF varnish. After the erosive challenge, the deposits were diminished in all groups. Conclusion Toothpastes containing 5wt.%, 20wt.%, and 30wt.% of S-PRG fillers behaved similarly to a conventional NaF toothpaste, even after an erosive challenge. The NaF varnish promoted better reduction of the Lp, but its effect was also diminished after erosion.

16.
Braz. J. Pharm. Sci. (Online) ; 58: e19473, 2022. tab, graf
Article in English | LILACS | ID: biblio-1384023

ABSTRACT

Abstract RGX-365 is the main fraction of black ginseng conmprising protopanaxatriol (PPT)-type rare ginsenosides (ginsenosides Rg4, Rg6, Rh4, Rh1, and Rg2). No studies on the antiseptic activity of RGX-365 have been reported. High mobility group box 1 (HMGB1) is recognized as a late mediator of sepsis, and the inhibition of HMGB1 release and recovery of vascular barrier integrity have emerged as attractive therapeutic strategies for the management of sepsis. In this study, we examined the effects of RGX-365 on HMGB1-mediated septic responses and survival rate in a mouse sepsis model. RGX-365 was administered to the mice after HMGB1 challenge. The antiseptic activity of RGX-365 was assessed based on the production of HMGB1, measurement of permeability, and septic mouse mortality using a cecal ligation and puncture (CLP)-induced sepsis mouse model and HMGB1-activated human umbilical vein endothelial cells (HUVECs). We found that RGX-365 significantly reduced HMGB1 release from LPS- activated HUVECs and CLP-induced release of HMGB1 in mice. RGX-365 also restored HMGB1-mediated vascular disruption and inhibited hyperpermeability in the mice. In addition, treatment with RGX-365 reduced sepsis-related mortality in vivo. Our results suggest that RGX- 365 reduces HMGB1 release and septic mortality in vivo, indicating that it is useful in the treatment of sepsis.


Subject(s)
HMGB1 Protein/analysis , Panax/adverse effects , Permeability , Sepsis/pathology , Ginsenosides , Human Umbilical Vein Endothelial Cells/classification , Anti-Infective Agents, Local/adverse effects
17.
Braz. J. Pharm. Sci. (Online) ; 58: e20007, 2022. tab, graf
Article in English | LILACS | ID: biblio-1394052

ABSTRACT

Abstract The prolonged entry of large amounts of calcium into the mitochondria through the mitochondrial calcium uniporter complex (MCUC) may cause the permeability transition pore (mPTP) to open, which contributes to the pathogenesis of several diseases. Tissue-specific differences in mPTP opening due to variable expression of MCUC components may contribute to disease outcomes. We designed this study to determine differential mPTP opening in mitochondria isolated from different regions of mouse brain and kidney and to compare it with the expression of MCUC components. mPTP opening was measured using mitochondria isolated from the left/right brain hemispheres (LH/RH, respectively) and from kidney cortex/medulla, while the expression level of MCUC components was assessed from total cellular RNA. Interestingly, LH mitochondria showed less calcium-induced mPTP opening as compared to RH mitochondria at two different calcium concentrations. Conversely, mPTP opening was similar in the renal cortex and renal medulla mitochondria. However, the kidney mitochondria demonstrated bigger and faster mPTP opening as compared to the brain mitochondria. Furthermore, asymmetric mPTP opening in the LH and RH mitochondria was not associated with the expression of MCUC components. In brief, this study demonstrates thus far unreported asymmetric mPTP opening in mouse brain hemispheres that is not associated with the mRNA levels of MCUC components.


Subject(s)
Animals , Male , Female , Mice , Brain , Calcium/agonists , Cerebrum/abnormalities , Mitochondrial Permeability Transition Pore/analysis , Mice , Mitochondria , 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine/adverse effects , Kidney Cortex
18.
Rev. nefrol. diál. traspl ; 41(4): 21-30, dic. 2021. graf
Article in Spanish | LILACS-Express | LILACS | ID: biblio-1377152

ABSTRACT

RESUMEN Objetivo: Analizar la asociación entre el polimorfismo rs2291166 del gen TJP1 con los niveles de glucosaminoglucanos (GAGS) excretados en orina como marcador de las primeras etapas de la nefropatía. Material y métodos: Se analizaron 600 muestras de orina de sujetos recién diagnosticados con diabetes tipo 2, de las cuales se incluyeron 203. La detección de GAGS en orina directa se realizó mediante prueba de turbidez de albúmina ácida y precipitación con cetilpiridinio (CPC). Resultados: El 26,64% de los pacientes diabéticos se encuentran en estadios tempranos de nefropatía, lo que corresponde a pacientes con prueba GAG positiva, siendo los que tienen mayor excreción de GAGS, heterocigotos para el polimorfismo. Conclusión:Sugerimos que el polimorfismo de TJP1 rs2291166 influye en la mucopolisacariduria en pacientes diabéticos tipo 2 de la población mexicana; que podría usarse como un marcador genético/ bioquímico válido para las primeras etapas de la nefropatía diabética.


ABSTRACT Objective: To analyze the association between the polymorphism rs2291166 of TJP1 gene, with the urine-excreted levels of GAGS as a marker of early stages of nephropathy. Methods: A 600 urine samples from newly diagnosed subjects with type 2 diabetes were analyzed, of which 203 were included. The GAGS detection in direct urine (corresponding to the first urine of the morning), was performed by albumin turbidity test and precipitation with cetylpyridinium (CPC). Results: The present study shows that 26.64% of diabetic patients are in early stages of nephropathy, corresponding to patients with a positive GAG test, being those with the highest GAGS excretion, heterozygous for the polymorphism. Conclusion: We suggest that the TJP1 polymorphism rs2291166 influences mucopolysacchariduria in type 2 diabetic patients of the Mexican population, which could be used as a valid genetic/biochemical marker for the early stages of diabetic nephropathy.

19.
Acta odontol. latinoam ; 34(1): 10-17, Apr. 2021. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1284929

ABSTRACT

ABSTRACT Dentin hypersensitivity is caused by increased dentinal permeability due to total or partial exposure of dentinal tubules, which in turn can be produced by alterations of dental structures or failure of restorative procedures. The purpose of this in vitro study was to evaluate the efficacy of the application of different kinds of adhesive systems to prevent dentin permeability before and after an erosive challenge. Fifty bovine dentin discs (6x1 mm) were prepared and the specimens were divided into 5 groups (n=10): (SB2) Single Bond 2, (SBU) Universal Single Bond, (CSB) Clearfil SE Bond, (SM) Scotchbond Multipurpose and (C) Control. Hydraulic conductance of dentin was recorded after adhesive application (HC-1) and after erosive challenge (HC-2). Dentin surface images of post-treatment and post-erosive challenge were obtained by scanning electron microscopy (SEM). Data were analyzed using Kruskal Wallis, Mann-Whitney with Bonferroni correction and Wilcoxon tests (p<0.05). Reduction in dentin permeability was observed with the application of adhesive systems (p<0.05). After the erosive challenge, dentin permeability increased for SBU and CSB (p<0.05), while SB2 and SM did not differ in HC-1 or HC-2 (p>0.05). The conventional, self-etching and universal adhesive systems reduce dentinal permeability by more than 80%, and dentin demineralization may contribute to the increased permeability of universal and self-etching systems.


RESUMO Diversos pacientes apresentam hipersensibilidade dentaria ou falha nos procedimentos restauradores, devido á exposigao total ou parcial de túbulos dentinários. O objetivo deste estudo in vitro foi o de avaliar a influencia da aplicagao de diferentes tipos de sistemas adesivos na permeabilidade da dentina e após o desafio erosivo. Cinquenta discos de dentina bovina (6x1 mm) foram confeccionados e os espécimes foram divididos em 5 grupos (n = 10): (SB2) Single Bond 2, (SBU) Universal Single Bond, (CSB) Clearfil SE Bond, (SM) ScotchbondMultiuso e (C) Controle. A condutáncia hidráulica da dentina foi registrada após a aplicagao do adesivo (HC-1) e após o desafio erosivo (HC-2). Imagens da superficie da dentina de pós-tratamento e pós-desafio erosivo foram obtidas por microscopia eletronica de varredura (MEV). Os dados foram analisados pelos testes de Kruskal Wallis, Mann-Whitney com correçâo de Bonferroni e Wilcoxon (p <0,05). Reduçâo da permeabilidade dentinária foi observada com a aplicaçâo dos sistemas adesivos (p <0,05). Após o desafio erosivo, a permeabilidade dentinária aumentou para SBU e CSB (p <0,05), enquanto SB2 e SM nao diferiram em HC-1 e HC-2 (p> 0,05). Os sistemas adesivos convencionais, autocondicionantes e universais reduzem a permeabilidade dentinária em mais de 80%, e a dentina desmineralizada pode gerar um aumento da permeabilidade dos sistemas adesivos universais e autocondicionantes.

20.
Article in Chinese | WPRIM | ID: wpr-909567

ABSTRACT

The fraction of horse chestnut seeds was named escins, which mainly consists of A, B, C, and D escin. Accumulating evidence suggests that escin exerts potent anti-inflammatory and anti-edematous effects. The effects of escin on inflammation and edema have been confirmed in various models. In a study in 1961, intravenous administration of escin was found to reduce acute edema in a rat paw. In the same study, escin was found to inhibit the increase in vascular permeability induced by egg white injection. Escin dose-dependently reduced the capillary permeability in chlo?roform-induced local inflammation in the abdominal skin surface of rabbits. The anti-inflammatory and anti-edematous effects of external use of escin were studied in carrageenan-induced paw edema and histamine-induced capillary perme?ability in rats. Escin gel decreased the contents of PGE2, TNF-α, and IL-1β, and reduced the raw edema and capillary permeability. The carrageenan-induced paw edema and pleuritis in bilaterally adrenalectomized rats were used to inves?tigate the anti-inflammatory effects of escin and glucocorticoid alone or combined. Co-administration of escin with corti?sone significantly reduced the volume of exudates and the number of white blood cells of exudates. The findings sug?gest escin can synergize with glucocorticoids to enhance their anti-inflammatory effect. The anti-inflammatory effect of escin was investigated in carrageenan-induced paw edema and acetic acid-induced capillary permeability in mice. Escin showed an anti-inflammatory effect, which is similar to that seen with dexamethasone treatment. However, escin showed a longer duration of the anti-inflammatory response than that of dexamethasone. Furthermore, escin had no signif?icant effects on spleen index, thymus index , proliferative capacity of splenocytes, lymphocyte count, and phagocytic rate. The findings suggest that escin is a potent anti-inflammatory drug with long-lasting anti-inflammatory effects without any immunosuppressive effects. Traditionally the mechanism of anti-inflammatory effect of escin is supposed to be rela?tive to release of PGF2α and corticosterone. The early studies showed that escin might promote the release of PGF2αand affect the pituitary adrenal system, stimulate the release of adrenocorticotropic hormone (ACTH) and glucocorticoid, which may explain its anti-inflammatory and anti-edema effects. Escin has glucocorticoid-like anti-inflammatory effect. However, escin did not exhibit an anti-inflammatory effect in low dose. Combination of suboptimal concentrations of escin with corticosterone inhibited the release of inflammatory factors including NO, TNF-αand IL-1βin the LPS-stimulated macrophage cells. Previous studies demonstrate that escin combined with glucocorticoid produced synergistic anti-inflammatory effects. The potential synergistic mechanisms may be associated with the property which escin regulates the glucocorticoid receptor (GR) signaling pathway. Escin can upregulate the expression of GR, promote the combina?tion of glucocorticoid and GR, then promote the activated GR transfer into the nucleus. Activated GR will inhibit the acti?vation of NF-κB directly, thus further inhibiting the expression of TNF-αand IL-1βand other inflammatory factors. Escin could inhibit 11β-HSD2 but not 11β-HSD1, thus decrease the metabolism of glucocorticoid. Escin and glucocorticoids have similar chemical structures. This indicate that one of the anti-inflammatory mechanisms of escin may be due to its stimulating GR by binding to it. Eacin might be a partial agonist of GR. A good many of researches have demonstrated the anti-inflammatory properties of escin, and shed light on the underlying mechanisms by which escin exerts these effects. Escin, as an oral or intravenous formulation, or a topical gel, inhibits inflammation, producing measurable improve?ments in edema and acute lung injury. Further clinical studies of escin are needed to demonstrate these properties in larger patient populations.

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