Uso de microvibración e inhibidores de la catepsina K en tratamientos de regeneración ósea dental / Use of micro-vibration and cathepsin K inhibitors in dental bone regeneration treatments

Introducción: el hueso está en remodelación constante para mantener la estructura del esqueleto, tener un ciclo de resorción por los osteoclastos y formación de hueso nuevo a cargo de los osteoblastos; el hueso también es susceptible a enfermedades sistémicas, traumas, edad y trastornos genéticos que afectarán el remodelado óseo, produciendo una pérdida masiva de masa ósea regulado por hormonas, citocinas, enzimas, etcétera. El objetivo es realizar una revisión sistemática de artículos que muestren cambio o alteración al utilizar tratamientos con microvibraciones y farmacológicos sobre la catepsina K en el hueso alveolar. Material y métodos: para realizar una comparación entre la efectividad del tratamiento a base de microvibraciones y con inhibidores de la catepsina K, se realizó una revisión sistemática en nueve bases de datos (Wiley Online Library, PubMed, Google Academic, Scopus, ScienceDirect, SciELO, Medline, EBSCO y Springer Link). La población de estudio fueron ratas y ratones. Resultados: en este estudio se incluyeron 20 artículos cuya investigación se realizó en estudios clínicos. En los resultados podemos observar cómo todos los tratamientos de alguna forma mejoran el proceso de remodelado óseo. Es difícil comparar cuál de los tratamientos dentro de cada grupo es mejor que otro, debido a que los resultados expresados son cualitativos. Conclusión: acorde a los resultados expresados se opta por realizar un tratamiento con microvibraciones debido a que el uso de inhibidores de la catepsina K aún no se encuentra completamente desarrollado y no se comprenden sus consecuencias debido a su manera sistémica de actuar (AU)

Introduction: the bone is in constant remodeling to maintain the skeletal structure, having a cycle of resorption by osteoclasts and formation of new bone by osteoblasts, the bone is also susceptible to systemic diseases, trauma, age and genetic disorders that affect bone remodeling, producing a massive loss of bone mass regulated by hormones, cytokines, enzymes, etcetera. The objective is to perform a systematic review of articles that show a change or alteration when using micro-vibration and pharmacological treatments on cathepsin K in the alveolar bone. Material and methods: in order to make a comparison between the effectiveness of micro-vibration and cathepsin K inhibitor treatments, a systemic review was carried out in nine databases (Wiley Online Library, PubMed, Google Academic, Scopus, ScienceDirect, SciELO, Medline, EBSCO and Springer Link). The study population was rats and mice. Results: this study included 20 articles whose research was carried out in clinical studies. In the results we can see how all the treatments in some way improve the bone remodeling process, it is difficult to compare which treatment within each group is better than the other, because the results expressed are qualitative. Conclusion: according to the results expressed, it is decided that it is better to perform a treatment with micro vibrations because the use of cathepsin K inhibitors are not yet fully developed and their consequences are not understood due to their systemic way of acting (AU)

Postoperative chronic pain in arthroplasty papillon-lefèvre syndrome: 17-year dental follow-up: case report / Síndrome de papillon-lefèvre: seguimiento dental de 17 años: reporte de caso

Introduction: The present report describes the case of a 12-year-old patient with 17-year follow-up who was previously diagnosed with Papillon-Lefèvre Syndrome (PLS), which is a rare autosomal recessive irregularity in the cathepsin C gene (CTSC) characterized by palmoplantar hyperkeratosis and premature loss of primary and permanent teeth. Case Report: A specific mutation in the c.203 T > G gene inducing loss of function leading to PLS was detected, as was a mutation in the HLA-DRB1*11 allele, which is associated with this syndrome. There is no consanguinity of the parents, and the siblings are entirely healthy. Early identification of the main characteristics of this syndrome is imperative. Accurate diagnosis by genetic analysis allows differential diagnoses and timely comprehensive dental treatment. Conclusions: Additionally, it allows consultation with a dermatologist to maintain or improve the quality of life of patients with this condition due to progressive worsening and severity of the main physical manifestations. Keywords: Papillon-Lefevre Disease; Keratoderma, Palmo-plantar; Cathepsin C; Periodontitis; Skin Diseases, Genetic; Case reports

Introducción: El presente reporte describe el caso de un paciente de 12 años de edad con 17 años de seguimiento a quien previamente se le diagnosticó Síndrome de Papillon-Lefèvre (PLS), el cual es una rara irregularidad autosómica recesiva en el gen de la catepsina C (CTSC) caracterizada por hiperqueratosis palmoplantar y pérdida prematura de dientes primarios y permanentes. Reporte de Caso: Se detectó una mutación específica en el gen c.203 T > G que induce pérdida de función que conduce a PLS, así como una mutación en el alelo HLA-DRB1*11, que se asocia a este síndrome. No presenta consanguinidad de los padres, padres y hermanos totalmente sanos. La identificación temprana de las principales características de este síndrome es imperativa. El diagnóstico certero por análisis genético permite diagnósticos diferenciales y tratamientos odontológicos integrales oportunos. Conclusiones: Adicionalmente, permite la consulta con un dermatólogo para mantener o mejorar la calidad de vida de los pacientes con esta condición debido al progresivo empeoramiento y severidad de las principales manifestaciones físicas.

Study on material basis of cathepsin K targeted inhibitor in Erzhi Wan / 药学实践杂志

Objective To investigate the active ingredients and components that inhibiting cathepsin K activity in Erzhi Wan, a classic kidney-tonifying formula. Methods Then-butanol, dichloromethane, ethyl acetate and petroleum ether parts and 30 active components in Erzhi Wan were screened by established high throughput fluorescence methods of inhibit the binding activity of CTSK with Z-FR-MCA substrate, the formation of CTSK and chondroitin sulfate A (CSA) complex activity, and the activity of substrate type I collagen degradation by CTSK. Molecular docking and insoluble collagen substrate binding assays were applied to verify the potential CTSK inhibitors. Results The n-butanol and petroleum ether parts of Erzhi Wan inhibited the formation of CTSK and CSA* complex by more than 90%, the petroleum ether part inhibited the binding of CTSK to substrate Z-FR-MCA by more than 90%, the collagen degradation inhibition rate of CTSK in n-butanol part was more than 95% and that in petroleum ether part was 58.6%. Among the 30 active components, 11 showed that the inhibition rate of CTSK and CSA* complex formation was more than 50%, and 5 components with the inhibition rate of Z-FR-MCA binding activity more than 50%. Finally, there were four components including eclalbasaponin Ⅸ, (-)-epicatechin gallate, nuezhenoside and wedelolactone. The inhibition rate of collagen degradation was more than 50%. Eclipta saponin IX inhibited the binding rate between collagen fibers and CTSK, up to 60%, but all of them failed to dock with CTSK active site. Conclusion There are active components that inhibiting cathepsin K in Erzhi Wan, which mainly exists in the n-butanol ingredients, but the active components is not an active-site inhibitor. It might inhibit the binding of CTSK with oligosaccharides by binding to other sites of CTSK, and then reduce the collagen degradation activity of CTSK.

Do matrix metalloproteinase and cathepsin K inhibitors work synergistically to reduce dentin erosion?

Abstract Objectives To evaluate the effects of matrix metalloproteinase (MMP) and cathepsin K (catK) inhibitors on resistance to dentin erosion. Methodology A total of 96 dentin specimens (3×3×2 mm) were prepared and randomly assigned into four groups (n=24): deionized water (DW); 1 µM odanacatib (ODN, catK inhibitor); 1 mM 1,10-phenanthroline (PHEN, MMP inhibitor); and 1 µM odanacatib + 1 mM 1,10-phenanthroline (COM). Each group was further divided into two subgroups for the application of treatment solutions before (PRE) and after erosive challenges (POST). All specimens were subjected to four daily erosive challenges for 5 d. For each erosive challenge, the specimens in subgroup PRE were immersed in the respective solutions before cola drinks, while the specimens in subgroup POST were immersed in the respective solutions after cola drinks (the immersion duration was 5 min in both cases). All specimens were stored in artificial saliva at 37°C between erosive challenges. The erosive dentin loss (EDL) was measured by profilometry. The residual demineralized organic matrix (DOM) of specimens was removed using type VII collagenase and evaluated by profilometry. Both the EDL and thickness of the residual DOM were statistically analyzed by two-way analysis of variance (ANOVA) and Bonferroni's test (α=0.05). The surface topography and transverse sections of the specimens were observed using SEM. MMPs and catK were immunolabeled in the eroded dentin and in situ zymography was performed to evaluate the enzyme activity. Results Significantly lower EDL was found in the groups ODN, PHEN, and COM than in the control group (all p<0.05), while no significant difference in EDL was found among the groups ODN, PHEN, and COM (all p>0.05). The application sequence showed no significant effect on the EDL of the tested groups (p=0.310). A significantly thicker DOM was observed in the group ODN than in the control group regardless of the application sequence (both p<0.05). The treatment with ODN, PHEN, and COM inhibited the gelatinolytic activity by approximately 46.32%, 58.6%, and 74.56%, respectively. Conclusions The inhibition of endogenous dentinal MMPs and catK increases the acid resistance of human dentin but without an apparent synergistic effect. The inhibition of MMPs and catK is equally effective either before or after the acid challenge.

New multienzymatic complex formed between human cathepsin D and snake venom phospholipase A2

Background Cathepsin D (CatD) is a lysosomal proteolytic enzyme expressed in almost all tissues and organs. This protease is a multifunctional enzyme responsible for essential biological processes such as cell cycle regulation, differentiation, migration, tissue remodeling, neuronal growth, ovulation, and apoptosis. The overexpression and hypersecretion of CatD have been correlated with cancer aggressiveness and tumor progression, stimulating cancer cell proliferation, fibroblast growth, and angiogenesis. In addition, some studies report its participation in neurodegenerative diseases and inflammatory processes. In this regard, the search for new inhibitors from natural products could be an alternative against the harmful effects of this enzyme. Methods An investigation was carried out to analyze CatD interaction with snake venom toxins in an attempt to find inhibitory molecules. Interestingly, human CatD shows the ability to bind strongly to snake venom phospholipases A2 (svPLA2), forming a stable muti-enzymatic complex that maintains the catalytic activity of both CatD and PLA2. In addition, this complex remains active even under exposure to the specific inhibitor pepstatin A. Furthermore, the complex formation between CatD and svPLA2 was evidenced by surface plasmon resonance (SPR), two-dimensional electrophoresis, enzymatic assays, and extensive molecular docking and dynamics techniques. Conclusion The present study suggests the versatility of human CatD and svPLA2, showing that these enzymes can form a fully functional new enzymatic complex.

Dano fotoinduzido no sistema endolisossomal: modulação da autofagia e metabolismo de lipídios em células de câncer colorretal / Endo-lysosomal system photodamage: modulation of autophagy and lipid metabolism in colorectal cancer cells

O câncer colorretal (CCR) é o terceiro câncer mais diagnosticado em humanos. O CCR causou mais de 900.000 mortes em 2020 e foi estimado, para o período entre 2020 - 2025, um incremento de 13.5 % no número de casos novos de acordo com a plataforma Web Global Cancer Observatory. A Terapia Fotodinâmica (PDT) é uma alternativa terapêutica promissora. Conhecer as vias de sinalização de morte celular, assim como, as respostas associadas com a resistência ao dano foto-oxidativo, são relevantes para incrementar a eficiência da PDT. Neste trabalho, investigamos como as células de adenocarcinoma colorretal HT 29 respondem ao dano fotoinduzido gerado pelo fotossensibilizador (FS) meso-tetrafenilporfirina dissulfônado (TPPS2a), uma molécula que é ativada pela irradiação com luz em 522 nm. Como esperado, após irradiação (2.1 J cm-2) foi verificado que com o incremento do TPPS2a houve diminuição da viabilidade celular. A concentração do FS escolhida para darmos seguimento ao estudo foi a necessária para reduzir em 30 % a sobrevida celular (DL30; 148 nM). Abordagens moleculares nos permitiram identificar que nas células fotossensibilizadas a redução na maturação da catepsina D (CTSD, 55 %) e da catepsina B (CTSB, 52 %) contribuem com a disfunção endolisossomal. Além disso, comprovamos que as células fotossensibilizadas tiveram, pela menor quantidade de CTSD ativa, o processamento da prosaposina (PSAP) significativamente afetado. Células coletadas após 24 horas de irradiação expressaram 7 vezes mais PSAP do que as amostras dos grupos controle, sugerindo que as reações de oxidação causadas pelo TPPS2a podem ocasionar o acúmulo de glicoesfingolipídios nos endossomos e nos lisossomos, mimetizando o fenótipo observado em doenças de armazenamento lisossomal. Imagens de células HT 29 com expressão estável da proteína LGALS3 fusionada ao marcador EFGP mostraram que, após 24 horas de irradiação, as células não ativaram a lisofagia para remover os endossomos e os lisossomos danificados. A ausência do recrutamento da LGALS3 também apontou que as membranas dos endossomos e dos lisossomos não apresentam rupturas permanentes que permitam a passagem de uma molécula de 26 kDa. Experimentos complementares de análise da expressão proteica dos marcadores autofágicos LC3-II e p62/SQSTM1 (referida como p62) confirmaram o bloqueio do fluxo autofágico nas células fotosenssibilizadas. Pelo envolvimento do sistema endolisossomal no tráfego de membranas e no fluxo de lipídios, o aumento da transcrição da Hidroximetilglutaril-CoA reductase (HMGCR) (≈ 1.6 vezes) uma enzima envolvida na síntese de novo do colesterol - sugeriu que a disfunção dos endossomos e dos lisossomos altera a distribuição de colesterol. Não obstante, para manter a homeostase lipídica nas células fotossensibilizadas este não foi o único mecanismocompensatório acionado, uma vez que houve um incremento sutil; porém, significativo (1.2 vezes) na transcrição da ceramidase ácida (ASAH1). Em conjunto, nossos dados apontam que a fotossensibilização com TPPS2a constitui uma ferramenta promissora para causar dano no sistema endolisossomal, inibindo a autofagia e permitindo o estudo das respostas metabólicas em células expostas a estresse oxidativo

Colorectal cancer (CRC) is the third most commonly diagnosed cancer in humans. CRC caused more than 900,000 deaths in 2020 and it was estimated for the period 2020 - 2025, an increase of 13.5 % in the number of new cases according to the Global Cancer Observatory Web platform. Photodynamic Therapy (PDT) is a promising therapeutic alternative. Understandings of cell death signaling pathways as well as the adaptive responses associated with resistance to photo-oxidative damage are relevant to optimize the effectiveness of PDT. For this purpose, in this research, we investigated how HT-29 colorectal adenocarcinoma cells respond to photosensitization reactions generated by TPPS2a, a molecule activated by irradiation with light at 522 nm. PS concentrations displayed increased inhibitory effect on cell viability after irradiation (2.1 J cm-2). The lethal dose selected to photosensibilize cells was the TPPS2a concentration able to reduce 30 % of cell survival (LD30; 148 nM). By molecular methods, we observed a reduction in cathepsin D (CTSD, 55 %) and cathepsin B (CTSB, 52 %) maturation, depletion that may contribute to endo-lysosomal dysfunction in photosensitized cells. It is widely known that endo-lysosomal cathepsins are crucial in protein turnover and degradation. Thus, we focused on the consequence of CTSD reduction. Literature data indicate that CTSD plays a key role in prosaposin (PSAP) processing to the four saposins (SAPs) that are required in glycosphingolipids breakdown. In fact, our results in photosensitized cells showed that, due to the lower amount of active CTSD, PSAP processing was significantly affected. Cells collected after irradiation expressed 7 times more PSAP than cells from the control groups. This data suggest that oxidative photodamage induced by TPPS2a may result in glycosphingolipid-accumulating endosomes and lysosomes, phenotype which mimics lysosomal storage diseases. Furthermore, we monitored by fluorescence microscopy a form of selective autophagy which detects and removes damaged endosomes and lysosomes known as lysophagy. Images of HT-29 cells expressing Galectin 3/LGALS3 fused to EFGP showed that photosensitized cells did not activate lysophagy. The absence of LGALS3 recruitment also indicated that the membranes of endosomes and lysosomes do not present ruptures which allow the passage of proteins with a molecular weight up to at least 26 kDa. Protein expression analysis of the autophagic markers LC3-II and p62/SQSTM1 (referred as p62) confirmed autophagic flux blockade in cells challenged with photoactivated TPPS2a. The endo-lysosomal system plays a key role in membrane trafficking and lipid flux. At the transcriptional level, 1.6-fold increase in gene expression of Hydroxymethylglutaryl-CoA reductase (HMGCR) - an enzyme involved in the synthesis de novo of cholesterol - indicated that endosomes and lysosomes dysfunction alters the distribution of cholesterol in cellschallenged with photoactivated TPPS2a. However, to maintain lipid homeostasis in photosensitized cells, this was not the only compensatory mechanism triggered, since there was a slightly increase (1.2-fold) in the transcription of acid ceramidase (ASAH1). Taken together, our data showed that photosensitization with TPPS2a constitutes a promising tool to damage the endolysosomal system, to inhibit autophagy and to study metabolic responses in cells exposed to oxidative stress

Role of cathepsin B in mechanical ventilator-induced lung injury in rats: relationship with NLRP3 inflammasomes / 中华麻醉学杂志

Objective:To evaluate the role of cathepsin B (CTSB) in mechanical ventilator-induced lung injury (VILI) in rats and the relationship with NOD-like receptor pyrin domain containing 3 (NLRP3) inflammasome.Methods:Thirty-six SPF-grade healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 220-300 g, were divided into 3 groups ( n=12 each) by the random number table method: control group (group C), VILI group (group V) and VILI + CA074-me group (group Me). CA074-me 5 mg/kg was intraperitoneally injected in group Me, while the equal volume of normal saline was given instead in group C and group V. Group C kept spontaneous breathing for 4 h, and the animals were mechanically ventilated (tidal volume 20 ml/kg, respiratory rate 80 breaths/min, fraction of inspired oxygen 21%, PEEP 0 cmH 2O). Blood samples from femoral artery were collected for arterial blood gas analysis before tracheal intubation and after spontaneous breathing or ventilation, and PaO 2 was recorded.Rats were sacrificed, and bronchoalveolar lavage fluid (BALF) was collected and lung tissues were collected for determination of the wet/dry lung weight ratio (W/D ratio), serum interleukin-1beta (IL-1β) and IL-18 concentrations in BALF (by enzyme-linked immunosorbent assay), expression of CTSB, NLRP3, apoptosis-associated speck-like protein containing a caspase-1 recruitment domain (ASC) and caspase-1 mRNA in lung tissues (quantitative real-time polymerase chain reaction), and expression of CTSB, NLRP3, ASC and caspase-1 in lung tissues (by Western blot) and for microscopic examination of the pathological changes (using HE staining). Lung injury was assessed and scored. Results:Compared with group C, PaO 2 was significantly decreased after the end of ventilation, the lung injury score, W/D ratio and concentrations of IL-1β and IL-18 in serum and BALF were increased, and the expression of CTSB, NLRP3, ASC and caspase-1 protein and mRNA in lung tissues was up-regulated in group V and group Me ( P<0.01). Compared with group V, PaO 2 was significantly increased after the end of ventilation, the lung injury score, W/D ratio and concentrations of IL-1β and IL-18 in serum and BALF were decreased, and the expression of CTSB, NLRP3, ASC and caspase-1 protein and mRNA in lung tissues was down-regulated in group Me ( P<0.01). Conclusions:CTSB is involved in VILI in the rats, and the mechanism may be related to activation of NLRP3 inflammasomes.

Selección de blanco terapéutico en la catepsina B para el desarrollo de fármacos contra el cáncer de mama / Therapeutic targeting of Cathepsin B for the development of drugs for breast cancer

RESUMEN Introducción: el cáncer de mama se ha incrementado en un 50 % en las dos últimas décadas. La catepsina B es una proteasa que participa en el proceso de tumurogénesis. Uno de los problemas actuales es la aparición de resistencias a fármacos. La búsqueda de nuevas alternativas terapéuticas puede reducir su morbimortalidad. Objetivo: caracterizar in silico estructural y funcionalmente la región conservada en la catepsina B como blanco terapéutico potencial en el tratamiento del cáncer de mama Métodos: con el uso de la herramienta ENTREZ del NCBI se obtuvieron 2 485 secuencias de la catepsina B. Las secuencias son sometidas a un alineamiento múltiple empleando Clustall Omega 1.2.4. Se realiza la caracterización estructural y funcional de la proteasa en estudio a partir de las bases de datos InterPro, Prosite, Uniprot y UniprotKB. Con el empleo del visualizador Jalview se seleccionó la mayor zona conservada de las especies de catepsina B. Resultados: la proteasa participa en la regulación de la actividad catalítica, proteólisis, regulación negativa de la muerte celular, procesos catabólicos del colágeno y posee actividad hidrolasa. El análisis múltiple permitió la visualización de las características aminoacídicas del sitio activo de la catepsina B y la selección de la región proteica más conservada. Conclusiones: la zona conservada de la catepsina B constituye un blanco potencial en el desarrollo de inhibidores como fármacos contra el cáncer de mama. Los análisis in silico reducen costo de las investigaciones actuales y contribuyen a la bioseguridad farmacológica.

ABSTRACT Introduction: breast cancer has increased by 50% in the last two decades. Cathepsin B is a protease involved in the process of tumorigenesis. One of the current problems is the emergence of drug resistance. The search for new therapeutic alternatives can reduce its morbidity and mortality. Objective: in-silico structural and functional characterization of the conserved region in Cathepsin B as a potential therapeutic target in the treatment of breast cancer. Methods: using the NCBI ENTREZ tool 2,485 Cathepsin B sequences were obtained. The sequences were subjected to multiple alignments using Clustall Omega 1.2.4. Structural and functional characterization of the protease under study was performed using the InterPro, Prosite, Uniprot and UniprotKB databases. Using the Jalview viewer, the largest conserved area of Cathepsin B species was chosen. Results: the protease is involved in the regulation of catalytic activity, proteolysis, negative regulation of cell death, collagen catabolic processes and possesses hydrolase activity. The multiple analyses allowed the visualization of the aminoacid characteristics of the active site of Cathepsin B and the selection of the most conserved protein region. Conclusions: the conserved region of Cathepsin B constitutes a potential target in the development of inhibitors as drugs against breast cancer. In-silico analysis reduces the cost of current research and contributes to pharmacological biosafety.

Decreased levels of cathepsin Z mRNA expressed by immune blood cells: diagnostic and prognostic implications in prostate cancer

Cathepsin Z (CTSZ) is a cysteine protease responsible for the adhesion and migration of both immune and tumor cells. Due to its dual role, we hypothesized that the site of CTSZ expression could be determinant of the pro- or anti-tumorigenic effects of this enzyme. To test this hypothesis, we analyzed CTSZ expression data in healthy and tumor tissues by bioinformatics and evaluated the expression levels of CTSZ mRNA in the blood cells of prostate cancer (PCa) patients by qRT-PCR compared with healthy subjects, evaluating its diagnostic and prognostic implications for this type of cancer. Immune cells present in the blood of healthy patients overexpress CTSZ. In PCa, we found decreased CTSZ mRNA levels in blood cells, 75% lower than in healthy subjects, that diminished even more during biochemical relapse. CTSZ mRNA in the blood cells had an area under the curve for PCa diagnosis of 0.832, with a 93.3% specificity, and a positive likelihood ratio of 9.4. The site of CTSZ mRNA expression is fundamental to determine its final role as a protective determinant in PCa, such as CTSZ mRNA in the blood cells, or a malignant determinant, such as found for CTSZ expressed in high levels by different types of primary and metastatic tumors. Low CTSZ mRNA expression in the total blood is a possible PCa marker complementary to prostate-specific antigen (PSA) for biopsy decisions, with the potential to eliminate unnecessary biopsies.

Effects of thrombolytic therapy time on serum inflammatory factor, cathepsin S, connective tissue growth factor and left ventricular ejection fraction in patients with acute ST-elevation myocardial infarction / 中国基层医药

Objective:To investigate the effects of thrombolytic therapy time on serum inflammatory factors, cathepsin S, connective tissue growth factor (CTGF), left ventricular ejection fraction (LVEF) and left ventricular end diastolic diameter (LVEDD) in patients with acute ST-elevation myocardial infarction.Methods:The clinical data of 119 patients with acute ST-elevation myocardial infarction who received thrombolytic therapy in the People's Hospital of Taierzhuang District of Zaozhuang from January 2019 to May 2020 were retrospectively analyzed. These patients were assigned to three groups according to different time points at which thrombolytic therapy was performed: group A (the time from onset to thrombolytic therapy ≤ 3 hours, n = 27), group B (3 hours < the time from onset to thrombolytic therapy ≤ 6 hours, n = 39), group C (6 hours < the time from onset to thrombolytic therapy ≤ 12 hours, n = 53). Recanalization rate, recanalization time, ST segment resolution rate at 2 and 12 hours, serum levels of inflammatory factors [including interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) and high-sensitivity C-reactive protein (hs-CRP)], cathepsin S, CTGF, LVEF, and LVEDD, and incidence of cardiovascular adverse events. Results:Recanalization time in group C was (148.73 ± 15.37) minutes, which was significantly longer than that in groups A and B [(89.34 ± 8.95) minutes, (98.76 ± 9.33) minutes]. Recanalization rate and ST segment resolution rate at 2 and 12 hours in group C were 45.28%, (40.17 ± 4.77) %, (73.92 ± 8.24) %, respectively, which were significantly lower than those in the groups A and B [96.30%, 79.49%, (47.42 ± 5.12)%; (83.68 ± 9.33)%, (43.56 ± 4.87)%, (78.73 ± 8.44)%] ( t/ χ2 = 248.088, 4.244, 20.204, 11.146, 18.508, 19.861, 6.271, 4.789, 17.995, 10.932, 3.339, 4.111, 4.100, 3.828, 3.100, 2.244, all P < 0.05). At 2 and 12 hours after thrombolytic therapy, IL-6, TNF-α and hs-CRP levels in group C were (23.29 ± 2.12) ng/L, (27.03 ± 2.75) ng/L, (6.49 ± 2.37) mg/L, (22.73 ± 2.05) ng/L, (26.24 ± 2.37) ng/L and (6.01 ± 2.53) mg/L, respectively, which were significantly higher than those in groups A and B ( t = 54.578, 54.578, 10.638, 8.584, 8.735, 5.199, 7.909, 7.171, 3.597, 1.382, 1.584, 1.008, 7.237, 5.190, 4.364, 8.829, 11.114, 2.585, 3.172, 6.815, 2.196, all P < 0.05). At 2 and 12 hours after thrombolytic therapy, cathepsin S and CTGF levels in group C were (29.97 ± 3.98) μg/L, (30.03 ± 4.79) μg/L, (28.05 ± 2.13) μg/L, (28.29 ± 4.31) μg/L, respectively, which were significantly higher than those in groups A and B [(31.74 ± 3.56) μg/L, (29.87 ± 4.91) μg/L; (20.81 ± 2.35) μg/L, (16.94 ± 3.46) μg/L; (30.95 ± 3.79) μg/L, (29.93 ± 4.95) μg/L; (26.37 ± 2.44) μg/L, (21.46 ± 4.79) μg/L, t = 93.870, 68.555, 15.039, 12.562, 6.345, 7.679, 3.096, 1.966, 13.882, 3.514, 11.863, 7.164, 9.239, 4.199, all P < 0.05). At 2 and 12 hours after thrombolytic therapy, LVEF and LVEDD in group C were (42.81 ± 4.77)%, (52.64 ± 4.71) mm, (43.13 ± 5.11)%, (51.57 ± 4.01) mm, respectively, which were significantly lower than those in groups A and B [(42.61 ± 4.58)%, (52.31 ± 4.47) mm, (46.33 ± 4.35)%, (47.75 ± 3.41) mm, (42.73 ± 4.79)%, (52.79 ± 4.76) mm, (44.79 ± 4.44)%, (49.93 ± 3.73) mm, t = 4.285, 9.193, 3.060, 4.214, 1.970, 2.953, 0.333, 1.259, 2.779, 1.626, 4.229, 1.996, 1.404, 2.416, all P < 0.05). The total incidence of cardiovascular adverse events was 7.41%, 12.82% and 33.96% in groups A, B and C, respectively ( χ2 = 4.383, all P < 0.05). Conclusion:The earlier the thrombolytic therapy time after acute ST-elevation myocardial infarction, the higher the recanalization rate and ST segment resolution rate, the milder the inflammatory reaction, atherosclerosis, the better the cardiac remodeling, the better the recovery of cardiac function, and the lower the incidence of cardiovascular adverse events.

Cathepsin D knockdown regulates biological behaviors of granulosa cells and affects litter size traits in goats / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Cathepsin D (CTSD), the major lysosomal aspartic protease that is widely expressed in different tissues, potentially regulates the biological behaviors of various cells. Follicular granulosa cells are responsive to the increase of ovulation number, hence indirectly influencing litter size. However, the mechanism underlying the effect of CTSD on the behaviors of goat granulosa cells has not been fully elucidated. This study used immunohistochemistry to analyze CTSD localization in goat ovarian tissues. Moreover, western blotting was applied to examine the differential expression of CTSD in the ovarian tissues of monotocous and polytocous goats. Subsequently, the effects of CTSD knockdown on cell proliferation, apoptosis, cell cycle, and the expression of candidate genes of the prolific traits, including bone morphogenetic protein receptor IB (

TRIB3 promotes B-ALL progression by suppressing CTSZ-mediated BCR-ABL degradation / 药学学报

Philadelphia chromosome (Ph) positive (Ph+) B cell acute lymphoblastic leukemia (B-ALL) is the most common genetic abnormality associated with B-ALL and has been shown to confer the worst prognosis to both children and adults. Increasing evidence has revealed that high tribbles homologue 3 (TRIB3) expression contributes to multi-cancer development and progression, but the underlying role and molecular mechanisms of TRIB3 in Ph+ B-ALL remain unclear. Here, we report that TRIB3 expression was enhanced in Ph+ B-ALL patient samples and positively associated with the expression of breakpoint cluster region-Abelson tyrosine kinase (BCR-ABL). We further demonstrated that deletion of TRIB3 attenuated the progression of Ph+ B-ALL by reducing BCR-ABL expression. Mechanistically, TRIB3 interacted with lysosomal cysteine proteinase cathepsin Z (CTSZ) to suppress CTSZ-mediated BCR-ABL degradation, which enhanced BCR-ABL activity, causing high proliferation of Ph+ B-ALL cells. Thus, our study indicated that inhibiting the expression of TRIB3 to regulate BCR-ABL kinase activity may be exploited as an additional target therapy for Ph+ ALL. Procedures for animal study were performed with approval of the Animal Care and Use Committee of the Chinese Academy of Medical Sciences and Peking Union Medical College. The procedure of human leukemia sample was approved by the Ethics Committee of Chinese Academy of Medical Sciences and Peking Union Medical College (KT2019055-EC-1).

Improvement of cognitive function in older adults based on near-infrared spectroscopy / 中国组织工程研究

BACKGROUND: Cognitive aging is an age-related cognitive degeneration that can develop into a cognitive dysfunction in the end stage. There is no clear therapeutic principle in clinical practice, but it is generally recognized that exercises can delay cognitive aging. OBJECTIVE: To summarize the current progress and shortcomings of exercise-delayed cognitive aging based on near-infrared spectroscopy. METHODS: In line with the PRISMA guidelines, the first author searched PubMed, WOS, CNKI, and WanFang using the keywords of “exercise, near-infrared spectroscopy, cognition, elderly or older adults, cathepsin B, brain-derived neurotrophic factor” in Chinese and English, respectively. Literature addressing senile cognitive function based on the near-infrared spectroscopy technique was retrieved, and 37 eligible articles were retained for further analysis. RESULTS AND CONCLUSION: We performed a comprehensive analysis of exercise-delayed cognitive aging based on near-infrared spectroscopy and confirmed that exercise improves the activation of cortex in different brain regions and thus improves cognitive function. Long-term aerobic exercise has better effects in the improvement of cognitive function than short-term exercise, which is more conducive to delaying cognitive aging. The underlying physiological mechanism may be that exercise improves blood flow in the brain, stimulates the secretion of neurotrophic factors from the skeletal muscle, and promotes the growth, survival and proliferation of neurons. However, there is no uniform standard for the interventional strategies. There are many links to be further improved, such as integration of individual differences and indexes in different brain regions, procedures for testing physical conditions (cardiovascular and lung diseases), to enhance the reliability of relevant parameters.

Alcohol extract of eucommia ulmoides oliv. Promotes bone healing in rats with periapical periodontitis / 中国组织工程研究

BACKGROUND: Current research on Eucommia ulmoides Oliv. mainly focuses on its use in the treatment of osteoarthritis that Eucommia ulmoides Oliv. can enhance the healing ability of bone tissue. However, research on its bone repair ability in periapical periodontitis has not been reported. OBJECTIVE: To study the effect of alcohol extract of Eucommia ulmoides Oliv. on cathepsin K expression in periapical periodontitis rats. METHODS: Twenty-four Sprague-Dawley rats were divided into normal control group (n=4) and apical periodontitis group (n=20). In the periapical periodontitis group, a periapical periodontitis model was established after exposure of the dental pulp in the first molar of the right mandible. The normal control group did not deal with any treatment. After 4 weeks of feeding, four rats from each group were taken for micro-CT detection. Bone destruction was quantified to confirm whether the rat model of periapical periodontitis was successfully constructed. After 5 weeks of feeding, the remaining 16 rats with periapical periodontitis were equally randomized into alcohol extract group (given alcohol extract of Eucommia ulmoides Oliv. via intragastric administration, 5 mL/kg per day) and normal saline group (given the same dose of normal saline via intragastric administration every day). After 4 weeks of gavage, four mice from each group were selected to perform micro-CT examination. The ability of alcohol extract of Eucommia ulmoides Oliv. to repair periapical bone tissue was analyzed. First molars of the right mandible from the other four rats in each group were extracted to detect the expression of cathepsin K in the alveolar bone using immunohistochemical staining. RESULTS AND CONCLUSION: Micro-CT results showed that the rat model of periapical periodontitis was successfully constructed as there was a significant difference in the bone resorption volume between the normal control and apical periodontitis groups [(0.223±0.009) mm3 vs. (0.945±0.037) mm3, P=0.00]. After 4 weeks of gavage, the micro-CT results showed that the alcohol extract of Eucommia ulmoides Oliv. significantly reduced the bone resorption volume in the rat model of periapical periodontitis (P < 0.05). Immunohistochemistry results showed that the alcohol extract of Eucommia ulmoides Oliv. significantly inhibited the expression of cathepsin K, a marker of bone destruction, in the rat model of periapical periodontitis. Therefore, these findings indicate that the alcohol extract of Eucommia ulmoides Oliv. can inhibit the expression of cathepsin K and promote the healing of bone tissue in the rats with periapical periodontitis.

Lysosomal Hydrolase Cathepsin D Non-proteolytically Modulates Dendritic Morphology in Drosophila / 神经科学通报·英文版

The main lysosomal protease cathepsin D (cathD) is essential for maintaining tissue homeostasis via its degradative function, and its loss leads to ceroid accumulation in the mammalian nervous system, which results in progressive neurodegeneration. Increasing evidence implies non-proteolytic roles of cathD in regulating various biological processes such as apoptosis, cell proliferation, and migration. Along these lines, we here showed that cathD is required for modulating dendritic architecture in the nervous system independent of its traditional degradative function. Upon cathD depletion, class I and class III arborization (da) neurons in Drosophila larvae exhibited aberrant dendritic morphology, including over-branching, aberrant turning, and elongation defects. Re-introduction of wild-type cathD or its proteolytically-inactive mutant dramatically abolished these morphological defects. Moreover, cathD knockdown also led to dendritic defects in the adult mushroom bodies, suggesting that cathD-mediated processes are required in both the peripheral and central nervous systems. Taken together, our results demonstrate a critical role of cathD in shaping dendritic architecture independent of its proteolytic function.

Estimation of bone mineral density and its correlations with homocysteine, and various other biochemical bone markers in postmenopausal women

Introduction- Omocysteine (HCY) prevents collagen cross-linking and activates osteoclast function within the bones. Bone mineral density (BMD) may be affected by Hyperhomocysteinemia via Cathepsin K. Aim- To find the correlation of BMD with biochemical bone markers. Methods- BMD was investigated by the DXA scan with the help of the Hologic QDR1000 system. As per WHO guidelines, subjects were divided into three different subsets with; normal bone mass, osteopenia, and osteoporosis. Every subject underwent routine biochemical laboratory investigations, HCY, Vitamin B12, and folic acid levels. Results-Among 355 postmenopausal women, 69% (245) had osteoporosis while 11.27% (40) had normal BMD (mean age, 53 ± 8.35 years) and 19.72% (70) had osteopenia (mean age 52.86 ± 7.93 years). The mean age in the osteoporotic group was 56.49 ± 6.65 years. The mean levels of HCY in the three groups were 15.58± 7.92 μmol/L, 16.13± 7.34μmol/L and 17.05± 5.13μmol/L, respectively. Hip BMD showed a strong inverse correlation with age (r=-0.360, p=0.002), while no significant correlations were found between weight and BMI. PTH was consistently seen to be negatively correlated with BMD at Spine (r=-0.0339, p=0.004), Forearm (r=-0.267, p=0.027), and Hip (r=-0.224, p=0.064). Conclusion- Low BMD is an important problem in postmenopausal female patients. Age and duration of menopause are independent risk predictors for the development of osteoporosis. Vitamin D levels do not predict low BMD in postmenopausal females. Weight is protective for osteoporosis especially at spine and forearm BMD. Vitamin B12 and Hcy levels did not correlate with low BMD.

New cathepsin V inhibitor from stems of Bowdichia virgilioides

Abstract Bioassay-guided fractionation of Bowdichia virgilioides Kunth, Fabaceae, extracts has led to the isolation of cathepsin V inhibitors. The investigation of the hexane and ethyl acetate extracts allowed the characterization of eleven compounds: lupeol, lupenone, β-sitosterol and stigmasterol in mixture, trans p-coumaric acid ester derivative, syringaresinol, bowdenol, 8-methoxycoumestrol, 3,4-hydroxy-7-methoxyisoflavone, 7,3′-dihydroxy-4′-methoxyisoflavone, and 5,4′-dihydroxy-7′-methoxyisoflavone. Structures of compounds were stablished by 1D and 2D NMR, and MS experiments. Among the isolated compounds, trans p-coumaric acid ester derivative and 8-methoxycoumestrol showed significant inhibition on cathepsin V, which is up to now unexplored.

The role of lysosomes in manganese-induced toxicity in SK-N-SH cells / 中华劳动卫生职业病杂志

Objective@#To investigate the role of lysosomes in manganese-induced toxicity in human neuroblastoma SK-N-SH cells.@*Methods@#SK-N-SH cells were treated with MnCl2 at doses of 0.062 5, 0.125, 0.25, 0.5, 1.0, 2.0 and 4.0 mmol/L for 24 h, and the cell viability was detected by MTT assay. Cells were treated with MnCl2 at doses of 0.125, 0.25, 0.5 and 1.0mmol/L for 24 h, and lysosomes labeled with lysotracker red were observed by laser confocal microscopy, the expression levels of LAMP1 and CTSD were detected by western blot, and CTSD activity was detected by Cathepsin D Activity Fluorometric Assay Kit.@*Results@#Compared with the control group, the survival rates of SK-N-SH cells were decreased significantly in the 0.5-4.0 mmol/L MnCl2 treatment groups (P<0.01) , the relative fluorescence intensities of 0.5 and 1.0 mmol/L MnCl2 treatment groups were increased (P<0.01) . Compared with the control group, the 0.125-0.5 mmol/L MnCl2 treatment groups had significant increase in the the expression of LAMP1 (P<0.01) . Compared with the control group, the expression of m-CTSD was significantly increased at the does of 0.125-0.25 mmol/L MnCl2, while it was decreased at the does of 1.0 mmol/L (P<0.01) . Otherwise, it wasn’t observed significant difference of the activity of CTSD between different MnCl2 treatment groups.@*Conclusion@#MnCl2 could cause cytotoxicity in SK-N-SH cells. Lysosomes may play a normal function at low doses of manganese, but they may be damaged at high doses of manganese. As an organelle that can degradate substrates in autophagy, lysosomes participate in the neurotoxic mechanism of manganese.

Progresses of cathepsin S in kidney and rheumatic diseases / 国际儿科学杂志

Cathepsin S(Cat S) is an important member of the cysteine cathepsin protease family.It is a lysosomal protease, which has been shown to be expressed in inflammatory conditions and autoimmune diseases.It has more specific roles such as antigen presentation, tissue repair, cell proliferation and apoptosis.It contributes to a variety of clinical diseases.In this review, we summarize the structure, characteristics and functions of cathepsin S in kidney and rheumatic diseases.

Pepstatin Pr show anti-fibrosis effect related to YAP-TGFβ-Smad pathway / 药学学报

Liver fibrosis is a tissue repair compensatory response to liver injury caused by various chronic factors, ultimately leading to liver cirrhosis, liver failure and even hepatocellular carcinoma. Abnormal activation of hepatic stellate cells is the cellular basis of liver fibrosis development. Pepstatin Pr, the derivative of pepstatin A, was isolated from Streptomyces sp. CPCC 202950. Our purpose was to investigate the anti-fibrotic activity of pepstatin Pr and explore its molecular mechanism. Hepatic stellate cell LX-2 was stimulated by TGFβ1 and sub- sequently treated with pepstatin Pr. Its cytotoxicity was detected by sulforhodamine B (SRB) assay. The expression of COL1A1, α-SMA and cathepsin D, signaling proteins TGFβ, Smad and YAP/TAZ were detected by Western blot or real-time PCR. The results showed that pepstatin Pr was not cytotoxic to LX-2 cells. And pepstatin Pr significantly reduced the mRNA and protein expression of COL1A1 and α-SMA, which are important liver fibrosis markers. Pepstatin Pr also repressed the protein expression level of cathepsin D, TGFβ1, YAP/TAZ, the phospholation level of Smad2, and YAP nuclear translocation. In conclusion, pepstatin Pr exhibits anti-fibrotic effects in TGFβ1-stimulaed LX-2 cells by mediating YAP-TGFβ-Smad pathway.