Bcl-2 Recombinant Rabbit Monoclonal Antibody

The sympathetic adrenergic nerves (SAN) play a significant role in the malignant transformation of breast cancer cells through the activity of norepinephrine (NE). However, the role of tyrosine hydroxylase (TH), a key enzyme of the NE synthesis, in the interaction between the SAN and triple-negative breast cancer (TNBC) cells has not been sufficiently explored, and whether TH can be a therapeutic target for TNBC has not been reported. TH expression in TNBC was examined by analyzing data from an online database and immunohistochemical analysis of our clinical samples. Cell proliferation and drug sensitivity were assessed upon coculture with fluorescently labeled cells using IncuCyte. Changes in the expression of DNA damage and apoptosis-associated proteins were assessed by western blotting. TH expression and NE synthesis in PC12 cells after their coculture with TNBC cells were measured by RT–qPCR, immunofluorescence, and ELISA. RNA sequencing was conducted on the cells before and after coculture. TH expression was relatively high in TNBC tumor tissues and closely associated with prognosis. SAN promoted TNBC cell proliferation through NE and reduced TNBC cell sensitivity to chemotherapeutic agents. Additionally, tumor cells induced TH expression in PC12 cells through nerve growth factor (NGF) secretion. Knocking down TH and using TH inhibitors effectively reversed the proliferation-promoting and drug sensitivity-reducing effects of SAN in TNBC cells. TH may be a central molecule in the positive feedback loop between TNBC cells and SAN. TH is a potential prognostic biomarker and can also serve as a therapeutic target for TNBC. Key messages The overexpression of the sympathetic nerve biomarker tyrosine hydroxylase (TH) in triple-negative breast cancer (TNBC) is an indicator for clinical staging, prognosis, and targeted therapy. There is a feedback loop where TH in nerves produces NE, worsening TNBC and reducing chemotherapy effectiveness. TNBC cell then increases TH expression, further boosting NE production. Furthermore, blocking the activity of TH could effectively inhibit this phenotype. Graphical

Endometriosis (EMS) is a chronic gynecological disease. RND3 is recognized as a potential autophagy-related biomarker in EMS. The aim of this study was to investigate the regulatory role of RND3 on autophagy and oxidative stress in EMS. Immunohistochemistry (IHC), RT-qPCR, and western blot (WB) analyses were used to determine the expression levels of RND3 and PLEKHG5. The study assessed oxidative stress by examining NRF2/NQO-1/HO-1 expression, as well as GSH, SOD, MDA levels, and lipid ROS production. Autophagy was evaluated by analyzing the expression of autophagy-related markers and phosphorylation of PI3K, AKT, ERK1/2, and mTOR. Cell proliferation, migration, and invasion were evaluated using CCK-8 and Transwell assays. Apoptosis was assessed through flow cytometry, expression of apoptosis-related markers, and TUNEL assay. The study also used ELISA to measure inflammatory factor levels and Co-IP assay to investigate the interaction between RND3 and PLEKHG5. Low expression of RND3 was observed in both the eutopic and ectopic endometrial tissues and ectopic endometrial stromal cells (EESCs) from patients with EMS. Increasing RND3 levels reduced oxidative stress in EESCs, enhanced cellular autophagy, inhibited cell proliferation, migration, and invasion, and promoted apoptosis. Conversely, the knockdown of RND3 expression had the opposite effect. The impact of RND3 overexpression on oxidative stress, autophagy, and apoptosis in EESCs was reversed by si-NRF2 and the autophagy inhibitor CQ. RND3 overexpression also upregulated the expression of PLEKHG5 in EESCs. Co-IP results revealed an interaction between RND3 and PLEKHG5. In in vivo experiments, low PLEKHG5 expression was observed in endometrial tissues of EMS mice, while RND3 overexpression alleviated EMS symptoms by decreasing oxidative stress and promoting cellular autophagy and apoptosis. RND3 inhibits EMS progression by enhancing autophagy and suppressing oxidative stress through PLEKHG5. Graphical RND3 alleviates endometriosis progression by suppressing oxidative stress and promoting autophagy, with its effects mediated through interaction with PLEKHG5.

Background Microglia-mediated neuroinflammation is closely related to the development of Alzheimer's disease (AD). This study further elucidated the regulatory mechanism of microglia polarization in AD. Method Microglia polarization was assessed using RT-qPCR, ELISA, and immunofluorescence (IF). Western blot (WB) analyzed inflammation-related, p-tau, and apoptosis-related proteins. Neuronal damage was evaluated by immunofluorescence, and neuronal apoptosis by flow cytometry and TUNEL assay. METTL3 and IκBα expression were detected using RT-qPCR and WB. N 6 -methyladenosine (m 6 A) levels were quantified with a colorimetric assay. RNA pull-down assay examined METTL3, IGF2BP2, and IκBα mRNA binding. IGF2BP expression was assessed by RT-qPCR. Learning and memory abilities were evaluated using morris water maze (MWM) test and novel object recognition (NOR) test. Inflammation-related proteins were detected using IF. Results Stimulation with Aβ 1-42 led to microglia M1 polarization, upregulation of inflammation-related proteins, and exacerbation of neuronal injury and apoptosis, along with increased p-tau expression in neurons. METTL3/IGF2BP2 modulated IκBα m 6 A modification through binding to IκBα mRNA, enhancing its expression. Enhanced METTL3 or IGF2BP2 expression suppressed M1 polarization, inflammation, and neuronal apoptosis in microglia, reversed by knockdown of IκBα. AD model mice exhibited cognitive impairments, neuroinflammation, and elevated M1 polarization. METTL3 or IGF2BP2 overexpression improved cognitive function, reduced neuroinflammation, and inhibited M1 polarization, and this effect was similarly reversed by knockdown of IκBα. Conclusion Our study demonstrates that the METTL3/IGF2BP2/IκBα axis is involved in neuroinflammation in AD by modulating microglia M1/M2 polarization, which sheds light on the treatment of AD.

Background Ferroptosis plays a key role in the development of chronic obstructive pulmonary disease (COPD). Whether ginsenoside Rg1 improves cigarette smoke-induced COPD or whether ginsenoside Rg1 improves COPD by inhibiting ferroptosis remains unknown. Methods BEAS-2B cells were exposed to cigarette solution (CSE) for 24 hours and treated with ginsenoside Rg1, the ferroptosis inhibitor Fer-1, and the PERK inhibitor GSK. Cell viability, endoplasmic reticulum stress, mitochondrial morphology, membrane potential, reactive oxygen species (ROS), iron levels, and the expression of related proteins were detected using corresponding methods. A COPD mouse model was constructed using cigarette smoke (CS). Ginsenoside Rg1 and GSK were administered via tube feeding 15 days after successful modeling. Mouse lung tissues were evaluated by HE staining. The expression of inflammatory markers, ROS, iron content, and related proteins was detected using corresponding methods. Results The results demonstrated that in the CSE-exposed BEAS-2B cell model and CS-induced mouse COPD model, the expression levels of endoplasmic reticulum stress (ERS)-related factors such as GRP78 were increased, while those of the antioxidant markers GPX4 and GSH were significantly decreased. Ginsenoside Rg1 improved emphysema and inflammation by inhibiting ferroptosis in vivo and in vitro. Using a PERK inhibitor, we found that ginsenoside Rg1 inhibited ferroptosis in vivo and in vitro by regulating ERS. Conclusion This study showed that ginsenoside Rg1 alleviates cigarette smoke-induced COPD by regulating the PERK/ATF4 axis to inhibit ERS and ferroptosis.