Tissue & cell最新文献

Background: Diabetic keratopathy is a prevalent but sometimes ignored visual condition in diabetic patients, which significantly affects patients with diabetes mellitus (DM) in terms of their visual acuity. Exosomes regulate diabetes-related conditions like diabetic keratopathy (DK) by secreting their components into the body.

Objective: Aim to investigate the effect and mechanism of mesenchymal stem cell (MSC)-derived exosome miR-125a-5p on DK.

Methods: Transmission electron microscopy, along with nanoparticle tracking analysis, was used to determine the morphology and size of exosomes. To evaluate cell viability, proliferation, and migration, Western blotting and RT-qPCR methods were used. CCK-8, cell cloning, and scratch assays were used to measure protein levels and mRNA expression.

Results: High glucose treatment of corneal epithelial cells weakened cell viability, proliferation and migration, and the level of miR-125a-5p was significantly reduced. It has been proposed that elevated levels of miR-125a-5p could enhance cell viability, proliferation, and migration, can inhibit endoplasmic reticulum stress induced by high glucose, which is the same as the effect of endoplasmic reticulum stress inhibitors.

Conclusion: Mouse bone marrow MSC-derived exosome miR-125a-5p repairs corneal epithelial cell viability and proliferation as well as migration ability to improve DK by inhibiting high glucose-induced endoplasmic reticulum stress.

Background: The objective of the present study was to assess potential immunotoxic effects of aspartame in BALB/c mice.

Methods: Aspartame was administered orally at 400 and 2000 mg/kg for two weeks (five days per week). Specific parameters of humoral and cellular immune responses including hemagglutinating antibody (HA) titer, cytokine production (IFN-γ and IL-4 levels), delayed type hypersensitivity (DTH) response to SRBCs, histopathological examination of spleen and bone marrow, and T-lymphocyte proliferation in response to phytohemagglutinin-A (PHA) were evaluated.

Results and conclusion: Aspartame at 400 and 2000 mg/kg did not significantly change hematological and histopathological parameters, HA titer, IFN-γ and IL-4 levels, DTH, and lymphoproliferation responses (p > 0.05). Aspartame at 400 and 2000 mg/kg did not induce any noticeable effects in immune system parameters of mice after a 14-day feeding. Aspartame was found to be safe to BALB/c mice immune system.

Podocyte injury is considered one of the main causes of kidney diseases occurrence and development. We have demonstrated that Ecto-5'-Nucleotidase (CD73) upregulated during podocyte injury, yet its function in podocyte is still unclear. Mouse podocytes cell line (MPC5) were exposed to the adriamycin (ADR, 0.25 μg/ml) to establish the model of podocytes injury, as well as low expression CD73 with lentivirus transfected shRNA. CD73 expression was verified by western blot and immunofluorescence assay. Cytokines (IL-1β, IL-18), apoptosis and apoposis-related protein (Bax, Caspase-3, Desmin) levels were measured using ELISA assay, Flow cytometry and Western blot, respectively. CD73, the cytokines of IL-1β and IL-18, apoptosis rate and the expression of Bax, Caspase-3 and Desmin were significantly increased in ADR group compared with the control group. Moreover, we also successfully constructed a CD73 down-expressed podocytes cell line. However, in comparsion with the ADR group, the cytokines of IL-1β and IL-18, apoptosis rate and the expression of Bax, Caspase-3 and Desmin protein were remarkably lowered in the ADR+CD73 shRNA group. These findings demonstrate that CD73 alleviates podocyte damage by reducing the inflammation and increasing apoptosis.

Objective: Investigate ADAM17 expression in colorectal cancer (CRC) at molecular and cellular levels and its potential mechanism in promoting tumorigenesis by regulating CRC cell migration and invasion.

Materials and methods: The study measured ADAM17 mRNA and protein levels in colorectal cancer cells and tissues using qPCR and immunohistochemical staining, and assessed the cells' proliferation, migration, and invasion abilities.

Results: ADAM17 expression was significantly higher in CRC tissues than in non-cancerous tissues and was linked to metastasis and poor prognosis in CRC patients. Silencing ADAM17 reduced cell migration and invasion. Mechanistically, knocking down ADAM17 decreased the expression of TGF-β/Smad pathway-related proteins, which inhibited proteins associated with migration and invasion, thus impairing these cellular processes.

Conclusion: ADAM17 likely promotes the migration and invasion of CRC cells by regulating the TGF-β/Smad signaling pathway. This study aids in understanding the molecular mechanisms of CRC metastasis and development, and supports the development of new therapeutic targets.

Male infertility is among one of the most challenging health concerns in the world. Traditional therapeutic interventions such as semen and testicular tissue cryopreservation aim to restore or preserve male fertility. However, these methods are subject to limitations that impact their efficacy and are infeasible in cases such as patients who cannot produce mature sperm due to genetic or pathological disorders. Moreover, with the number of cases of prepubertal boys who must undergo gonadotoxic treatments rising, alternatives have been sought for fertility preservation to enhance reproductive rates in vitro and in vivo. Tissue engineering is a promising area that can address aspects that current therapies may not fully encompass through the creation of bioartificial testicular structures or 3D culture systems that allow the establishment of the essential conditions for sperm production. This study aims to first give a brief overview of stem cell therapy in treating male infertility and then go more in-depth regarding the novel methods and procedures based on tissue engineering that have the potential to offer new treatments for infertility caused by testicular disorders and defects.

Objective: Bone resorption mediated by osteoclast differentiation induces the occurrence of bone-related diseases. Macrophages, an origin of osteoclasts, whose M2 type can reduce inflammation-induced bone damage. We aimed to investigate the effect of M2 macrophage-derived exosomes on osteoclast formation and elucidate its underlying mechanism.

Materials and methods: Exosomes were isolated from M2 macrophages (M2-exo) and were used to treat osteoclast-like cells. Osteoclast formation was evaluated using tartrate-resistant acid phosphatase, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blotting. The molecular mechanism of M2-exo function was analyzed by qRT-PCR, phosphor-kinase array analysis, and Western blotting.

Results: M2-exo was internalized by osteoclasts and inhibited osteoclast differentiation in vitro. Moreover, CYLD was highly expressed in M2 macrophages and M2-exo-treated osteoclasts, and knockdown of it abrogated the inhibition of osteoclast differentiation caused by M2-exo. Additionally, CYLD suppressed the phosphorylation of STAT3, and STAT3 activator colivelin reversed the inhibition of osteoclast differentiation induced by CYLD overexpression.

Conclusion: M2-exo inhibits osteoclast differentiation via delivering CYLD, which inactivates STAT3 signaling. These findings may provide a novel therapeutic option for bone diseases including periodontitis.

Dysregulation in placental trophoblast cells frequently results in oxidative stress, culminating in pregnancy-related complications. While iron is essential for fetal development, cellular ferroptosis due to elevated iron levels might mediate the emergence of preeclampsia (PE), presenting significant risks during gestation. We found abnormally activated oxidative stress and increased iron concentration in the placental tissues of PE patients. Subsequently, we treated placental trophoblasts with hydrogen peroxide and RSL3 to induce oxidative stress and ferroptosis models. The results revealed that SIRT6 overexpression activates the Nrf2/HO-1 pathway, restores the oxidative imbalance of the cells, and protects the cells from ferroptosis. Meanwhile, activation of the Nrf2/HO-1 pathway alone showed similar results. Thus, we posit that SIRT6, via the Nrf2/HO-1 pathway, alleviates cellular oxidative stress and diminishes ferroptosis, offering a novel therapeutic avenue for PE.

The ezrin/radixin/moesin proteins play a central role in cross-linking plasma membrane proteins with the actin cytoskeleton. Despite intensive ERM protein research in many tissues and pathologies, little is known about these proteins in healthy tissues of reproductive organs. Therefore, we examined ezrin, phosphorylated ezrin/radixin/moesin (pan-pERM), radixin, and moesin distribution at the cellular level by means of immunohistochemistry in ovaries of the following animal species: mouse, dog, cat, sheep, pig, horse, and cynomolgus monkey. Ezrin was expressed in oocytes, ovarian surface, granulosa cells and corpus luteum. A characteristic, predominantly membranous pan-pERM staining pattern was observed in ovarian surface epithelium, oocyte, granulosa cells and corpus luteum. Moesin immunoreactivity was present in all ovarian structures with a prominent signal in endothelial cells of blood vessels. Oocytes, granulosa cells and corpus luteum revealed mainly nuclear radixin staining. Staining pattern and subcellular localization (membranous, cytoplasmic, nuclear) varied between different animal species and between particular ERM proteins as well. This data may help gain new insights into the physiological function of ERM proteins in biological events in the female reproductive system.

Asthma is a prevalent chronic pediatric lung disease which is commonly perceived as a syndrome of airway inflammation characterized by cough and wheeze in clinic. Klotho is implicated in diverse cellular activities, including inflammation, oxidative stress and apoptosis. This paper aims to explore the role of klotho in asthma and investigate the relevant molecular reaction mechanisms. To this end, we used Der p1 to induce an in vitro asthma model in BEAS-2B cells. Klotho expression was manipulated in Der p1-induced BEAS-2B cells with overexpression and its effects on Der p1-induced pathologies including apoptosis and inflammatory cytokine levels and the expressions of oxidative stress-related markers and major mediators in endoplasmic reticulum stress (ER stress) were investigated. Mitochondrial membrane potential (MMP) and mitochondrial permeability transition pore (mPTP) opening were also detected. Our data demonstrated that Der p1 stimulation decreased klotho expression and klotho overexpression inhibited the Der p1-induced inflammation, oxidative stress and apoptosis. Overexpressing klotho inhibited ER stress to modulate mitochondrial function. The inhibitory effects of klotho overexpression were reversed by ER stress agonist tunicamycin. This paper validated the role of klotho in asthma pathogenies and developed prospective therapeutic targets for asthma treatment.