Platelet Microparticle-Derived MiR-320b Inhibits Hypertension with Atherosclerosis Development by Targeting ETFA.

Hypertension and atherosclerosis often occur simultaneously. This study aimed to explore the role and mechanism of platelet microparticle (PMP) -derived microRNA-320b (miR-320b) in patients with hypertension accompanied by atherosclerosis.We collected samples from 13 controls without hypertension and atherosclerosis and 20 patients who had hypertension accompanied by atherosclerosis. In vitro, platelets were activated by Thrombin receptor-activating peptide to produce PMPs. HUVECs were induced by CoCl2 to mimic a hypoxic environment in vitro. RT-qPCR was employed to detect the expression levels of CD61, miR-320b, and ETFA. The protein expression level of ETFA was evaluated via Western blotting. Furthermore, 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide, 5-ethynyl-2'-deoxyuridine, and wound healing assays were employed to assess the proliferation and migration of HUVECs. Enzyme-linked immunosorbent assay was used to measure the oxidative stress and inflammation-related factor expression.The expression of miR-320b was reduced in both platelets and PMPs but increased in plasma. MiR-320b promoted CoCl2-induced HUVEC viability, proliferation, and migration. The levels of the oxidative stress factors SOD and GSH as well as the inflammatory factor IL-10 were elevated in the CoCl2 + miR-320b mimics group compared with both the CoCl2 + mimics NC and CoCl2 groups. Conversely, the levels of the oxidative stress factors MDA and ROS as well as the inflammatory factors IL-6, TNF-α, and IL-1ß were decreased. These results were regulated by miR-320b targeting ETFA.PMP-derived miR-320b inhibits the development of hypertension accompanied by atherosclerosis by targeting ETFA.

A multidisciplinary study on Clinostomum infections in Nile tilapia: micro-morphology, oxidative stress, immunology, and histopathology.

Yellow grub disease, caused by Clinostomum metacercaria, is an endemic zoonotic infection in freshwater fish, responsible for Halzoun syndrome transmitted through the consumption of raw infected fish. This study aimed to conduct a multidisciplinary investigation integrating detailed morphology, oxidative stress, immunology, and histopathology alteration to advance our understanding of Clinostomum infection. In this annual study, 400 Nile tilapia (Oreochromis niloticus) were collected from the Nile River at Al Bahr Al Aazam, Giza Governorate to assess Clinostomum infection prevalence. Of the examined fish, 160 individuals (40.0%) harboured larval Clinostomum infections. Clinostomum metacercariae were observed in various anatomical locations, with 135 fish (33.8%) in buccal cavities, 21 fish (5.25%) in gill chambers, and 4 fish (1.0%) on the skin. Infection intensity ranged from 2 to 12 cysts per fish, averaging 5 cysts, notably with skin infections characterized by a single cyst in each fish. Macroscopic encysted metacercariae were collected from buccal cavities, gills, and skin. Micro-morphology revealed distinct features in C. complanatum, including an elliptical oral sucker with collar-like rings and large sensory papilla-like structures, contrasting with the absence of these features in C. phalacrocoracis. Oxidative stress, assessed through malondialdehyde (MDA) and nitric oxide levels, revealed an elevation in MDA to 35.13 ± 6 nmol/g and nitric oxide to 25.80 ± 3.12 µmol/g in infected fish. In infected fish, MHC-I gene expression increased approximately 13-fold, MHC-II peaked at 19-fold, and IL-1ß significantly upregulated by 17-fold, compared to control levels. Histopathology detailed associated lesions, such as cyst encapsulation and eosinophilic infiltration. Clinstomiasis and its impacts on fish hosts offer crucial insights to control this emerging fish-borne zoonotic disease, threatening wildlife, aquaculture, and human health.

Exploring the mechanism of Liuwei Dihuang formula for promoting melanin synthesis in juvenile zebrafish based on network pharmacology and molecular docking.

Background: Vitiligo stands as a challenging skin disorder with limited treatment options available. LiuWei DiHuang formula (LDF), a renowned Traditional Chinese medicine, has exhibited promising results in treating vitiligo over an extended period. However, the precise underlying mechanism of its action remains elusive. Methods: Employing a comprehensive network pharmacology approach, this study identified active compounds and their corresponding targets within LDF, while also pinpointing vitiligo-associated targets sourced from the TCMSP database, OMIM, DisGenNET, and Genecards. A network was established to illustrate the connections between active compounds and targets, alongside a protein-protein interaction network. Further analyses, encompassing Gene Ontology (GO) function and KEGG pathway enrichment, were conducted using the DAVID platform. Molecular docking simulations were performed utilizing AutoDockTools and AutoDockVina software. To validate the outcomes of the systematic pharmacological investigation, experiments were conducted using juvenile zebrafish. Results: The collective effort of the network pharmacology approach yielded a compilation of 41 compounds and 192 targets. Molecular docking simulations notably revealed the lowest binding energies for CAT-quercetin and CAT-Kaempferol interactions. The utilization of juvenile zebrafish experiments highlighted a significant increase in melanocyte count following methoxsalen and LDF treatment. Notably, LDF prominently augmented the expression levels of proteins related to melanogenesis. Additionally, LDF showcased the capacity to enhance CAT and SOD levels while concurrently reducing ROS and MDA activity. In contrast to the model group, substantial increases in protein and mRNA levels of Nrf2 and HO-1 were observed in response to LDF treatment (P < 0.05). Conclusion: Through a meticulous network pharmacology approach, this study successfully predicted active components and potential targets associated with LDF's application in vitiligo treatment. The therapeutic effectiveness of LDF against vitiligo is postulated to stem from its regulation of oxidative stress factors and the Nrf2/HO-1 pathway.

Serum oxidative stress factors predict myocardial ischemia reperfusion injury after percutaneous coronary intervention in patients with acute myocardial infarction and type 2 diabetes mellitus.

Introduction: Serum oxidative stress factors may be considered to be essential parameters for indicating cell oxidative damage. Aim: We designed this study to investigate the clinical diagnostic value of serum oxidative stress factors (superoxide dismutase (SOD), malondialdehyde (MDA), and myeloperoxidase (MPO)) combined with ischemia-modified albumin (IMA) and heat shock protein 70 (HSP70) for myocardial ischemia-reperfusion injury (MIRI) after percutaneous coronary intervention (PCI) in elderly patients with acute myocardial infarction (AMI) and type 2 diabetes mellitus (T2DM). Material and methods: From November 2020 to August 2021, 94 patients with AMI + T2DM and 86 patients with AMI were enrolled in the study; they were sub-grouped into the MIRI and non-MIRI groups following the occurrence of MIRI within 48 h after PCI. SOD, MDA, MPO, IMA, and HSP70 levels were determined. The clinical values of the combined serum oxidative stress factors, IMA, and HSP70 levels to predict MIRI events were analyzed. Results: There was a higher probability of MIRI events in the AMI + T2DM group than the AMI group (p < 0.05). The ROC curve for the combined prediction of SOD, MDA, MPO, IMA, and HSP70 for the occurrence of MIRI events was higher in both the AMI and the AMI + T2DM groups than for predictive factors alone (all p < 0.05). Conclusions: Combined prediction of SOD, MDA, MPO, IMA, and HSP70 has the highest diagnostic value for predicting MIRI events after PCI in AMI patients, especially in patients with AMI combined with T2DM.

Effect of “lumbar three needles” on expression of oxidative stress factors in rats with multifidus muscles injury / 中国组织工程研究

BACKGROUND: Posterior lumbar spine surgery is currently the main surgical approach for many surgical operations such as discectomy and spinal canal decompression. However, 10%-40% of patients with posterior lumbar spine surgery can have low back pain and related dysfunctions soon after surgery, which is related to excessive stretching or blunt injury to the paraspinal muscles such as the multifidus muscle during the operation. OBJECTIVE: To explore the effect of “Lumbar Three Needles” on the expression of oxidative stress factors, including malondialdehyde, reactive oxygen species, superoxide dismutase, glutathione peroxidase and phosphorylated protein kinase B, in rats with multifidus muscles injury. METHODS: Twenty-four male Sprague-Dawley rats were randomly divided into control group, model group and “Lumbar Three Needles” group, with 8 rats in each group. An animal model of multifidus muscles injury was made by intramuscular injection of 0.5% bupivacaine hydrochloride in the model and treatment groups in control and “Lumbar Three Needles” group, while normal saline was injected in the control group. No acupuncture intervention was given in the control group and model group, while Dachangshu, Shenshu and Weizhong acupoints were electroacupunctured in the “Lumbar Three Needles” group. Needles were then stimulated electrically using a Han's Acupoint Nerve Stimulator with density wave, frequency of 2 Hz/10 Hz and continuous current of 1 mA for 20 minutes, once daily for 7 days in total. The multifidus muscle was stained with hematoxylin-eosin staining to observe morphological changes, and kits were used to observe the expression of malondialdehyde, reactive oxygen species, superoxide dismutase, and glutathione peroxidase, while western blot was applied to observe the expression of phosphorylated protein kinase B. The study protocol was approved by the Animal Ethic Committee of Guangdong Second Traditional Chinese Medicine Hospital with an approval No. 048617. RESULTS AND CONCLUSION: Hematoxylin-eosin staining results indicated that the muscle fibers were partially fixed in the model group and there were still a lot of macrophages after 7 days of intervention. Compared with the model group, more newborn muscle fibers and fewer macrophages could be seen in the “Lumbar Three Needles” group. After intervention, the cross-sectional area of the multifidus muscles in the “Lumbar Three Needles” group was significantly bigger than that in the model group (P < 0.01). The expression levels of malondialdehyde and reactive oxygen species in the model group were significantly increased compared with the control group (P < 0.01) and “Lumbar Three Needles” group (P < 0.01). The expression of superoxide dismutase, glutathione peroxidase and phosphorylated protein kinase B in the “Lumbar Three Needles” group was significantly higher than that in the model group (P < 0.01 or P < 0.05). Therefore, “Lumbar Three Needles” can significantly decrease the oxidative stress level after multifidus muscle injury, which may be related to the improvement of phosphorylated protein kinase B.

Investigating the effect of intracameral cefuroxime on oxidative stress and apoptosis in the rat cornea / Investigando o efeito de cefuroxima intracameral sobre o estresse oxidativo e a apoptose na córnea de ratos

ABSTRACT PURPOSE: We examined the effect of intracameral administration of cefuroxime on oxidative stress and endothelial apoptosis in rat corneal tissue. METHODS: In total, 30 rats were divided into 3 groups of 10 rats each (intracameral administration of cefuroxime 0.1 mg/0.01 mL (cefuroxime group); intracameral administration of balanced salt solution 0.01 mL (control group); or absence of intracameral injection (sham group). Corneal endothelial apoptosis was assessed by immunohistochemical analysis using caspase-3 and caspase-8. Total oxidant status, total antioxidant status, oxidative stress index, and paraoxonase and arylesterase levels were examined in corneal endothelial tissue and serum. RESULTS: Paraoxonase levels in the serum were significantly different between the sham and cefuroxime groups (p=0.027). A significant difference was also observed in total oxidant status levels between the cefuroxime and balanced salt solution groups (p=0.023). In addition, there were significant differences in total antioxidant status levels in corneal tissue between the cefuroxime and sham groups (p<0.001) and between the cefuroxime and balanced salt solution groups (p<0.001). Furthermore, significant differences were also observed in oxidative stress index levels between the cefuroxime and balanced salt solution groups (p=0.001) and between the cefuroxime and sham groups (p=0.026). According to the immunohistochemical staining results, a significant association with caspase-3 activity existed between the cefuroxime and balanced salt solution groups (p=0.007), while no significant difference was found with caspase-8 activity (p=0.541). Caspase-3 activity exhibited a significant relationship between the sham and balanced salt solution groups (p=0.018), but no relationship was found with caspase-8 activity (p=0.623). CONCLUSION: Immunohistochemical examination revealed that intracameral cefuroxime increased apoptosis when compared to the sham and balanced salt solution groups. Moreover, intracameral cefuroxime increased oxidative stress in the cornea and simultaneously induced apoptosis.

RESUMO OBJETIVO: Examinamos o efeito da administração intracameral da cefuroxima sobre o estresse oxidativo e a apoptose endotelial no tecido corneano de ratos. MÉTODOS: No total, 30 ratos foram divididos em 3 grupos de 10 ratos cada (administração intracameral de cefuroxima 0,1 mg/0,01 mL (grupo cefuroxima), administração intracameral de solução salina balanceada 0,01 mL (grupo controle) ou ausência de injeção intracameral (grupo sham)). A apoptose endotelial da córnea foi avaliada por análise imuno-histoquimica usando caspase-3 e -8. O status oxidante total, o status antioxidante total, o índice de estresse oxidativo e os níveis de a paraoxonase e arilesterase foram investigados no tecido endotelial da córnea e no soro. RESULTADOS: Os níveis de paraoxonase no soro foram significativamente diferentes entre os grupos sham e cefuroxima (p=0,027). Foi também observada uma diferença significativa nos níveis de estado oxidante total entre os grupos cefuroxima e solução salina balanceada (p=0,023). Além disso, houve diferenças significativas nos níveis de status antioxidante total no tecido da córnea entre os grupos cefuroxima e sham (p<0,001) e entre os grupos cefuroxima e solução salina balanceada (p<0,001). Diferenças significativas também foram observadas nos níveis do índice de estresse oxidativo entre os grupos cefuroxima e solução salina balanceada (p=0,001) e entre os grupos cefuroxima e sham (p=0,026). De acordo com os resultados de coloração imuno-histoquimica, houve associação significativa com a atividade da caspase-3 entre os grupos cefuroxima e solução salina balanceada (p=0,007), enquanto não houve diferença significativa com a atividade da caspase-8 (p=0,541). A atividade da caspase-3 exibiu uma relação significativa entre os grupos sham e solução salina balanceada (p=0,018), mas nenhuma relação foi encontrada com a atividade da caspase-8 (p=0,623). CONCLUSÃO: O exame imuno-histoquímico revelou que a cefuroxima intracameral aumentou a apoptose quando comparada com os grupos sham e solução salina balanceada. Além disso, a cefuroxima intracameral aumentou o estresse oxidativo na córnea e induziu simultaneamente a apoptose.

Impaired Skeletal Muscle Regeneration Induced by Macrophage Depletion Could Be Partly Ameliorated by MGF Injection.

Skeletal muscle injury is one of the most common injuries in sports medicine. Our previous study found that macrophage depletion impairs muscle regeneration and that mechano growth factor (MGF) may play an important role in this process. However, whether injection of MGF protects against impaired muscle regeneration after macrophage depletion has not been explored. Therefore, we generated a muscle contusion and macrophage depletion mouse model and injected MGF into the damaged muscle. Comprehensive morphological and genetic analyses were performed on the injured skeletal muscle after macrophage depletion and MGF injection. The results showed that injection of MGF did not exert a protective effect on muscle fiber regeneration; however, it did decrease fibrosis in the contused skeletal muscle after macrophage depletion. Moreover, MGF injection decreased the expression of muscle inflammatory cytokines (TNF-α, IFN-γ, IL-1ß, and TGF-ß), chemokines (CCL2, CCL5, and CXCR4), oxidative stress factors (gp91phox) and matrix metalloproteinases (MMP-1, MMP-2, MMP-9, MMP-10, and MMP-14). These results suggest that the impairment of skeletal muscle regeneration induced by macrophage depletion could be partly ameliorated by MGF injection and that inflammatory cytokines, oxidative stress factors, chemokines, and MMP may be involved in this process.

Changes in inflammatory and oxidative stress factors and the protein synthesis pathway in injured skeletal muscle after contusion.

Injury of skeletal muscle, and particularly mechanically-induced damage, including contusion injury, frequently occurs in contact sports as well as in sports with accidental contact. Although the mechanisms of skeletal muscle regeneration are well understood, those involved in muscle contusion are not. A total of 40 male mice were randomly divided into control (n=8) and muscle contusion (n=32) groups. A muscle contusion model was established by weight-drop injury. Subsequently, the gastrocnemius muscles in the two groups were harvested at different times (1, 3, 7 and 14 days) post-injury. The changes in skeletal muscle morphology were assessed by hematoxylin and eosin (H&E) stains. Furthermore, quantitative polymerase chain reaction and western blotting were used to analyze inflammatory cytokines, oxidative stress factors and the Akt/mechanistic target of rapamycin (mTOR) pathway. The results revealed that pro-inflammatory cytokines [tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interferon-γ (IFN-γ)] increased significantly at day 1 and 3 and still exhibited high levels of expression at days 7 and 14 (except IL-6) post-injury. Additionally, the anti-inflammatory cytokine IL-10 increased significantly at 1, 3 and 7 days and reached its peak levels at 7 days post-injury. It was revealed that gp91phox mRNA increased significantly at all time points and gp91phox protein increased significantly at day 3 post-injury. Furthermore, it was observed that p-Akt/Akt increased significantly at 1 day post-injury. P-mTOR/mTOR increased significantly at day 1 and 7, and p-p70s6k/p70s6k and P-4EBP1/4EBP1 increased significantly at 1, 3, 7 and 14 days post-injury. These results indicate that inflammatory and oxidative stress factors and the Akt/mTOR pathway may serve important roles in the regeneration of muscle contusion. In addition, certain inflammatory factors and oxidative stress factors maintained high levels of expression at 14 days after injury, indicating that the healing process of muscle was still not fully achieved at this time.