Molecular Medicine最新文献

The use of recombinant growth factors has led to improved hematopoietic function after chemotherapy, but hematologic and immune complications still occur after chemotherapy which can be life-threatening. Here we show that pan-SHIPi compounds can induce endogenous G-CSF and TPO production in vivo and can also increase host survival after a lethal fungal challenge. In addition, we show that consistent with their ability to induce G-CSF, pan-SHIPi compounds can promote increased granulopoiesis in normal mice and speed recovery of neutrophil counts after chemotherapy. We also report the identification of a novel SHIP1-selective inhibitor, A32, that also increases steady state production of both G-CSF and TPO. These findings indicate small molecule inhibitors of SHIP1 can promote hematologic recovery after myeloablative chemotherapy and are a unique means to induce endogenous production of multiple growth factors that promote hematologic recovery.

Background: Sevoflurane is known to induce cognitive dysfunction, but the underlying mechanisms remain unclear. Recent evidence suggests that disruptions in synaptic zinc homeostasis may contribute to neurotoxicity and cognitive impairment. This study investigates the role of synaptic zinc imbalance in sevoflurane-induced cognitive dysfunction and evaluates the neuroprotective effects of estrogen.

Methods: Aged female C57BL/6 mice were exposed to sevoflurane to induce neurotoxicity. Synaptic zinc levels, Tau phosphorylation, synaptic vesicle numbers, neuronal firing frequency, and neuronal damage were assessed. The effects of zinc chelation with CaEDTA and estrogen supplementation on these parameters, as well as cognitive performance in the Morris water maze and Y-maze tests, were evaluated.

Results: Sevoflurane exposure disrupts synaptic zinc homeostasis by upregulating Znt3 expression, leading to increased Tau phosphorylation, reduced synaptic vesicle numbers, decreased neuronal firing frequency, elevated neuronal death, and cognitive impairment. Chelation of zinc with CaEDTA attenuated Tau phosphorylation and neuronal death, enhanced neuronal firing, and improved cognitive function. Estrogen supplementation alleviates synaptic zinc imbalance by downregulating Znt3 expression, thereby reducing Tau phosphorylation and neuronal loss, increasing synaptic vesicle density and neuronal firing frequency, and improving cognitive function.

Conclusions: This study reveals that sevoflurane-induced cognitive dysfunction is closely associated with synaptic zinc imbalance. Estrogen exerts its neuroprotective effects by restoring synaptic zinc homeostasis. These findings provide insights into the pathophysiological mechanisms underlying anesthesia-related cognitive impairment and highlight the therapeutic potential of estrogen in perioperative neuroprotection.

The study reported here offers new insights into the metabolic changes associated with the Warburg effect (i.e. aerobic glycolysis) in cancer cells and into the possible role of IF₁, the endogenous inhibitor of ATP synthase that preserves cellular energy when it works in reverse, hydrolyzing ATP. We investigated biochemical and main bioenergetic parameters in cell lines derived from three human tumors: osteosarcoma (143B), colon carcinoma (HCT116), and cervix carcinoma (HeLa). The combination analysis of cellular glucose consumption, lactate production, ATP-linked respiration rate, ATP level, cell culture medium acidification rate, and ROS level demonstrates that aerobic glycolysis is differently expressed by the three different types of tumor cells, although all cell types exhibited a Warburg phenotype. The superoxide anion level was found to be lower in HCT116 cells, which showed the highest ratio between oxidative phosphorylation and glycolysis rates, while ROS level was similar in all cells examined, suggesting that mitochondria in HCT116 are very efficient in both energy production and limiting their oxidative stress. Additionally, IF₁ KD cells of all kinds of tumor showed higher level of ROS compared to their related IF₁-expressing cells. Most of the results reported here clearly demonstrate that aerobic glycolysis is completely independent on both the level of IF₁ and the IF₁/ATP synthase ratio, excluding the contribution of an IF₁-dependent mechanism in the metabolic shift of cancer cells towards glycolysis. Indeed, the study provides a detailed analysis of the bioenergetics of tumor cells exhibiting very different IF₁/ATP synthase ratios and shows that IF₁ KD cells of all tumor types had a higher level of ROS than their related IF₁-expressing cells. Overall, the comprehensive picture of tumor cell bioenergetics would facilitate the identification of appropriate drugs for targeted tumor treatments, such as ATP synthase-IF₁ immunotherapy that would strongly limit cellular resistance to severe hypoxia or anoxia, where IF₁ plays an effective critical role.

Background: Radiation enteritis (RE) is a common complication in patients undergoing abdominal and pelvic radiotherapy. Despite the advancements in radiotherapy, effective treatments remain limited. WGX50, a bioactive compound from Sichuan pepper, has shown anti-inflammatory and antioxidant properties. This study investigates the protective effects of WGX50 on RE, focusing on its potential to reduce radiation-induced damage in the intestine.

Methods: Network pharmacology and molecular docking were used to identify the molecular targets of WGX50. In vitro, human intestinal epithelial cells (HIEC6) and colon cells (NCM460) were exposed to radiation and treated with WGX50. In vivo, C57BL/6 mice were administered WGX50 prior to radiation exposure. Various assays, including CCK-8, colony formation, flow cytometry, histopathology, and 16S rRNA sequencing, were performed to evaluate cell proliferation, apoptosis, oxidative stress, intestinal damage, and gut microbiota composition. Tissue transcriptome sequencing was conducted to explore differentially expressed genes.

Results: In vitro, WGX50 significantly mitigated radiation-induced cell damage, enhanced cell proliferation, and reduced apoptosis at non-toxic concentrations. In vivo, WGX50 treatment preserved intestinal morphology and reduced inflammatory infiltration in irradiated mice. WGX50 also protected goblet cells, maintaining mucin production and epithelial barrier function critical for intestinal homeostasis. Molecular docking, dynamics simulations and surface plasmon resonance (SPR) revealed stable binding of WGX50 to Epidermal Growth Factor Receptor (EGFR), key targets involved in oxidative stress regulation and ferroptosis inhibition. Mechanistically, WGX50 upregulated the EGFR-SLC7A11-GPX4 axis, suppressing ferroptosis and protecting intestinal cells. Additionally, 16S rRNA sequencing showed that WGX50 mitigated radiation-induced gut microbiota dysbiosis, preserving microbial diversity and promoting beneficial bacterial populations.

Conclusion: WGX50 demonstrates potent radioprotective effects by reducing oxidative stress, suppressing ferroptosis, and maintaining intestinal homeostasis, including goblet cell function and gut microbiota composition. These findings support WGX50's potential as a novel therapeutic agent for the prevention and treatment of radiation enteritis.

Background: Neutrophils are critical first responders of the innate immune system, rapidly recruited to sites of infection or sterile injury. Upon activation by pathogen- or damage-associated molecular patterns, neutrophils initiate antimicrobial responses, including cytokine release, phagocytosis, and the formation of neutrophil extracellular traps (NETs). While NETosis plays a protective role, excessive NET formation can exacerbate inflammation and tissue damage. Pulmonary exposure to ricin, a potent toxin derived from Ricinus communis, results in acute lung injury characterized by neutrophil infiltration, cytokine production, vascular leakage, and pulmonary edema. This study investigated the contribution of NETosis to ricin-induced lung pathology and explored the therapeutic potential of targeting NETosis with a long acting recombinant DNase I (PRX-119) to attenuate lung injury.

Methods: CD1 outbreed mice were pulmonary exposed to ricin, and bronchoalveolar lavage fluid (BALF) and lung tissues were collected at various time points post-exposure. NETosis was assessed by immunofluorescence and Western blot analysis of markers, including peptidyl arginine deiminase 4 (PAD4), citrullinated histone H3 (citH3) and myeloperoxidase (MPO). Therapeutic intervention included administration of a NET-degrading DNase agent in combination with an anti-ricin antibody. Cell-free DNA levels, NETosis markers, neutrophil infiltration, lung histopathology, vascular permeability and the expression of pro- and anti-inflammatory mediators were evaluated. Weight loss and survival were also monitored and compared between anti-ricin monotherapy and combined anti-ricin and plant-produced human recombinant long acting (LA) DNase I (PRX-119), a novel NET degradation therapy.

Results: Ricin exposure led to elevated pulmonary levels of PAD4, citH3 and MPO, accompanied by extensive NET formation in both BALF and lung tissue. Mice receiving combined therapy with a newly developed DNase I - based agent (PRX-119) and an anti-ricin antibody treatment exhibited significantly improved survival and reduced weight loss compared to antibody monotherapy. The combined treatment not only significantly reduced NETosis markers, but also improved lung histopathology, reduced vascular leakage and pulmonary edema and altered the levels of proteins involved with pro- or anti-inflammatory effects, Dkk-1, CD93 and Periostin.

Conclusions: These findings demonstrate for the first time that NETosis plays a significant pathological role in ricin-induced lung injury. Moreover, they underscore the therapeutic potential of combining advanced NET-degrading agents, specifically PRX-119, an advanced DNAse I under development, with toxin-neutralizing antibodies as a promising strategy to reduce acute lung damage and enhance clinical outcomes.

Recurrent pregnancy loss (RPL) profoundly impacts not only the physical health but also the psychological well-being of women. Despite its profound effects, the underlying pathophysiological mechanisms of RPL remain largely elusive, with few discernible warning signs. Lymphocyte activation gene-3 (LAG-3) is a crucial immune checkpoint that modulates immune responses during infection and tumor. In the present study, we examined the expression of LAG-3 on CD4⁺T cells during pregnancy via cytometry by time-of-flight and flow cytometry. Our findings revealed a higher frequency of LAG-3⁺ decidual CD4⁺T (dCD4⁺T) cells in response to trophoblasts during normal pregnancy. This specific LAG-3⁺ subset of dCD4⁺T cells was found to produce a greater number of anti-inflammatory cytokines. Notably, blocking LAG-3 was highly effective in inhibiting the production of anti-inflammatory cytokines, which is detrimental to the maintenance of pregnancy. A decrease in the number of LAG-3⁺dCD4⁺T cells was correlated with miscarriage. Interestingly, the RNA level of LAG-3 (data analyzed from the two published single-cell databases) remained stable in RPL. Palmitoylation might play a role in regulating LAG-3 expression during RPL, as the palmitoylation of LAG-3⁺dCD4⁺T cells was increased in RPL. Additionally, the general palmitoylation inhibitor 2-bromopalmitate was found to upregulate LAG-3 expression on dCD4⁺T cells both in vitro and in vivo. Collectively, these findings highlighted the significant roles of LAG-3 in regulating the function of dCD4⁺T cell and maintaining normal pregnancy. Furthermore, they suggested that lower LAG-3 expression on dCD4⁺T cells could serve as a potential biomarker for diagnosis of RPL.