Journal of Physiology and Pharmacology最新文献

Myopia is one of the most prevalent refractive errors and one of the leading causes of visual impairment and blindness worldwide. It results from a mismatch between the axial length and optical power of the eye, resulting in a focal plane that lies in front of the retina. In children and young adults, myopia is most commonly caused by excessive elongation of the eyeball during development - a hallmark of school-age and some early-onset genetic forms of myopia. However, myopic refractive error can also result from other mechanisms, such as increased lens power in age-related nuclear cataracts or corneal steepening in keratoconus, which are not associated with axial elongation. The prevalence of myopia in young Asian adults has increased from 20-30% to 80-85% over the last 50 years. In contrast, recent meta-analytic data for European young adults, emphasizing studies with cycloplegic refraction essential for accuracy, indicate myopia prevalence rates of approximately 19-24%. The prevalence of high myopia (greater than or equal to-6.0 diopters) has increased disproportionately to myopia in the last 50 years, from 1-5% to 10-20% and became a global problem. The reason for this state of affairs is believed to be lifestyle and prolonged near vision activities. Although refractive error can be corrected, sight-threatening pathologies such as retinal detachment, macular degeneration, glaucoma, and cataracts are more challenging to control. Owing to years of research, the biological mechanisms of eye growth and refractive development are increasingly elucidated. The signaling cascade mechanisms that link the retinal image processing and alterations in choroidal thickness and scleral development have also been studied. While the retina can detect defocus and changes in defocus, decades of research have led to a growing understanding of the fundamental pathways in visually guided eye growth, yet the precise initial mechanisms by which the retina senses and transduces these optical signals continue to be an active and important area of investigation. Animal studies have demonstrated that the retina can locally regulate visually guided eye growth through intrinsic mechanisms, even in the absence of direct input from the brain. The precise molecular mechanisms underlying common forms of myopia, particularly those involving axial elongation, are yet to be fully elucidated. This reflects the complexity and multifactorial influences inherent even in these prevalent forms, alongside the challenges posed by experimental models in completely recapitulating all aspects of the human condition.

Gastric cancer (GC) is one of the most prevalent malignant cancers, with currently unsuccessful treatment strategies for patients. Increased PI3K/AKT/STAT-3 pathway activity has been observed in patients with GC. Morin (MRN), which is a flavonoid, exhibits significant anticancer activity by inhibiting the PI3K/AKT signaling pathway. However, monotherapy with MRN has faced challenges due to poor bioavailability and rapid elimination. This study investigated the combined effects of MRN and paclitaxel (PTX) on apoptosis induction and their molecular mechanisms in GC cells (HGT-1). After 24 h of MRN and PTX exposure, various assays were performed to assess the suppression of HGT-1 cell proliferation. These included cytotoxicity assessments, reactive oxygen species (ROS) level measurements, apoptotic morphological features, mitochondrial membrane potential (ΔΨm), nuclear fragmentation, and cell cycle analysis. Further, the effect of MRN and PTX on STAT-3 expression and various proliferation and apoptotic proteins was investigated using western blotting. The results revealed that the MRN and PTX combination significantly induced cytotoxicity, increased ROS levels, and altered ΔΨm, resulting in HGT-1 cell apoptosis (P<0.05). Furthermore, MRN and PTX treatment decreased the expression of oncogenic proteins, such as C-Fos, KRAS, and p-ERK1, in HGT-1 cells (P<0.05). The combination treatment inhibited PI3K, AKT, and STAT3 expressions, thereby suppressing proliferation and inducing proapoptotic protein expression in HGT-1 cells. Therefore, the combination of MRN and PTX could serve as a therapeutic approach for malignant GC treatment.

Patients with Crohn's disease (CD) often exhibit low serum zinc levels. Experimentally, colitis models induced by intrarectal injection of 2,4,6-trinitrobenzenesulfonic acid (TNBS) have shown exacerbated pathogenesis with zinc deficiency. Zinc, which is regulated by transporters (zip and znt), plays a crucial role in maintaining intestinal integrity. Zip transports zinc into the cytoplasm from the extracellular spaces or organelles, while znt moves it out of the cytoplasm into the extracellular space or into organelles. Although CD is an intractable gastrointestinal inflammatory condition that is developed predominantly in the terminal ileum, few studies have focused on the ileitis. Therefore, we aimed to identify zinc transporters related to ileitis using the TNBS/ethanol-induced ileitis model. Ileitis was induced in mice by ileal luminal injection of TNBS (1.5 mg/0.8 mL/mouse) in 50% ethanol. The mice were euthanized 6, 24, or 48 h after the injection for histological and physiological analyses. Serum zinc levels and the expression of zinc transporters in duodenal and ileal tissues were also evaluated. Ileal luminal injection of TNBS/ethanol significantly increased tumor necrosis factor alpha (TNF-α) and interleukin 1beta (IL-1β) mRNA expression at 24 h, then caused ileal epithelium detachment at 48 h. Serum zinc levels were significantly decreased, accompanied by reduced expression of zip4 and znt1 in the ileum, 6 and 24 h after the injection. The expression of zip4 and znt1 in the duodenum was significantly increased at 24 and 48 h, respectively, and serum zinc levels recovered to around normal levels. Furthermore, zip7 expression in the ileum significantly decreased at 24 h but recovered slightly at 48 h, coinciding with re-epithelialization. Overall, the pathogenesis of TNBS/ethanol-induced ileitis involves dynamic changes in zinc transporters.

The roles of circular RNAs (circRNAs) in the concurrent processes of mitochondrial damage and angiogenesis following cerebral ischemia-reperfusion (CI/R) injury remain largely uncharacterized. This study investigated the function of circUCK2 in CI/R and tested the hypothesis that it exerts a protective effect by modulating the miR-188-5p/phosphatase and tensin homolog (PTEN) signaling axis. In vivo middle cerebral artery occlusion/reperfusion (MCAO/R) and in vitro oxygen-glucose deprivation/reperfusion (OGD/R) models were employed. A combination of lentiviral delivery, neurological scoring, histological analysis (hematoxylin and eosin, TUNEL), RT-qPCR, Western blot, flow cytometry (apoptosis, reactive oxygen species), tube formation assays, and dual-luciferase reporter assays was used to investigate the underlying mechanisms. In CI/R models, circUCK2 and PTEN expression was significantly downregulated, while miR-188-5p was upregulated (P<0.05). Overexpression of circUCK2 in MCAO/R mice significantly ameliorated neurological deficits, attenuated neuronal apoptosis, mitigated mitochondrial oxidative stress, and promoted angiogenesis (P<0.05 for all outcomes). Mechanistically, circUCK2 was identified as a cytoplasmic sponge for miR-188-5p, which in turn was shown to directly target and suppress PTEN expression (P<0.05). Crucially, the neuroprotective and pro-angiogenic effects of circUCK2 were significantly counteracted by PTEN knockdown (P<0.05), positioning PTEN as a critical downstream mediator of circUCK2's function. We found that circUCK2 exerts a protective role against CI/R injury by sequestering miR-188-5p, thereby derepressing PTEN expression. This study identifies the circUCK2/miR-188-5p/PTEN axis as a key regulatory pathway and a promising therapeutic target for ischemic stroke.

Triple-negative breast cancer (TNBC) lacks effective therapeutic targets. This study investigated the mechanism by which circular RNA eukaryotic translation initiation factor 6 (circEIF6) regulates TNBC progression via the miR-296-3p/metallothionein 2A (MT2A) axis. Tumor and adjacent tissues from 32 TNBC patients were analyzed. TNBC (MD Anderson Metastatic Breast-231) and non-TNBC (MCF-7) cells were used to assess circEIF6, miR-296-3p, and MT2A expression. Functional assays measured glycolysis (including glucose uptake, lactate production, adenosine triphosphate levels, extracellular acidification rate/oxygen consumption rate), angiogenesis (anti-vascular endothelial growth factor A/fibroblast growth factor-2 expression, human umbilical vascular endothelial cell tube formation), and cell proliferation. Target interactions between circEIF6, miR-296-3p and MT2A were validated by luciferase/RNA immunoprecipitation assay. CircEIF6 was significantly upregulated in TNBC (P<0.05), correlating with advanced TNM (Tumor, Node, Metastasis) Classification of Malignant Tumors stage (P=0.01), tumor size (P=0.01) and poor prognosis. Silencing circEIF6 suppressed glycolysis, angiogenesis, and proliferation (P<0.05) by sponging miR-296-3p to downregulate MT2A. We conclude that circEIF6 promotes aerobic glycolysis and angiogenesis in TNBC via the miR-296-3p to target MT2A.

Caffeine is a well-known compound broadly consumed by humans as an ingredient in beverages. Its ingestion is associated with numerous beneficial effects on the organism. Caffeine may, among others, reduce adipose tissue accumulation. However, the mechanism underlying this action is poorly elucidated. Lipolysis is a relevant process directly related to adipocyte metabolism. The present study explored the short-term (2-h) effects of 1 mM caffeine on the lipolytic activity of isolated adipocytes. Lipolysis was measured as the quantity of glycerol released from cells. The expression of lipolysis-related genes was also determined at the mRNA level. It was shown that caffeine significantly increased lipolysis induced by epinephrine (an adrenergic receptor agonist), forskolin (a direct activator of adenylate cyclase (AC)), and dibutyryl-cAMP (a direct stimulator of protein kinase A (PKA)). However, the lipolytic response of fat cells to DPCPX (an adenosine A₁ receptor antagonist) was unchanged by caffeine. Moreover, caffeine failed to affect lipolysis stimulated by the combination of DPCPX with epinephrine, DPCPX with DB-cAMP and DPCPX with forskolin. Additionally, the stimulatory effect of caffeine on epinephrine-induced glycerol release was suppressed by H-89 (a direct inhibitor of PKA). Caffeine slightly increased the cAMP content (a signaling molecule increasing the lipolytic process) in adipocytes. However, the expression of analyzed genes was not significantly affected in the presence of the tested compound. The results show that caffeine promotes lipolysis in primary rat adipocytes by inhibiting the adenosine A1 receptor signaling pathway. The pharmacological suppression of PKA abolishes the stimulatory effect. Increased lipolysis in the presence of caffeine contributes to reduced lipid accumulation in adipocytes.

Ulcerative colitis (UC) belongs to inflammatory bowel disease (IBD), a group of chronic disorders of the gastrointestinal (GI) tract with unknown etiopathogenesis. The prevalence of IBD is increasing, and available therapeutic options do not lead to complete remission; therefore, there is a need to identify novel pharmaceutical strategies for UC. The aim of our study was to evaluate the suitability of the PSB-KK-1415, the G protein coupled receptor 18 (GPR18) receptor agonist, in the treatment of intestinal inflammation in mice. The impact of PSB-KK-1415 on the viability of human epithelial cell line CCD 841 CON was assessed using MTT assay. Dextran sodium sulfate (DSS) was used to induce colitis in mice. Colonic samples were collected at different stages of disease and Gpr18 expression was determined using real time RT-PCR. In the DSS model, the mice were treated with PSB-KK-1415 (5 mg/kg once daily) by different routes of administration. The anti-inflammatory potential of PSB-KK-1415 in vivo was evaluated based on macroscopic and microscopic scoring systems. As a result PSB-KK-1415 did not affect the viability of CCD 841 CON cells after 24 and 48 hours. The mRNA expression of GPR18 was significantly increased in the early and moderate stages of colitis as compared to the control group and decreased in severe colitis and during recovery. In the acute mouse model of UC, 5 mg/kg PSB-KK-1415 injected intraperitoneally tended to reduce the area of inflammation and improved the intestinal wall thickness as compared to mice with colitis. PSB-KK-1415 decreased MPO activity. No significant differences in anti-inflammatory effect were found with regard to the route of administration. In the chronic model of DSS-induced colitis, PSB-KK-1415 demonstrated anti-inflammatory properties, evidenced by a decreased area of inflammation in the colon and improved intestinal thickness as compared to inflamed animals. Our findings indicate that activation of the GPR18 receptor may be a potential target for modulation the course of chronic colitis in mice.

Pentobarbital (PB), a barbiturate anesthetic, is widely used in many clinical treatments, including management of seizures and preoperative sedation. However, its potential role in cancer therapy remains underexplored. The inhibitory effects of PB on breast cancer proliferation were investigated in this study. Breast cancer cell lines MDA-MB-231 and MCF-7 were used to evaluate the effects of PB on cell proliferation. Proliferating cell nuclear antigen (PCNA) overexpression and knockdown models were established to assess its role in PB-mediated proliferation inhibition. Cell proliferation was measured using methyl thiazolyl tetrazolium (MTT), colony formation, and bromodeoxyuridine (BrdU) incorporation assays. Protein and mRNA expression levels of PCNA and cell cycle-related genes were analyzed by Western blot and real-time RT-PCR, respectively. Statistical analysis was performed using Student's t-test or one-way ANOVA. PB was found to inhibit cell proliferation and regulate the expression of cell cycle-related genes compared to the control group. Further analysis revealed that PB downregulated the expression of PCNA. Overexpression of PCNA increased the proliferation of MDA-MB-231 cells, while PCNA knockdown suppressed it. Notably, overexpression of PCNA could partially restore the proliferative capacity of MDA-MB-231 cells that had been inhibited by PB. The findings indicate that PB, in addition to its established roles as a sedative and anesthetic agent, suppresses breast cancer proliferation through the downregulation of PCNA expression. These results provides new theoretical evidence supporting the potential application of PB in cancer treatment.

Vaspin, a visceral-adipose-tissue-derived serine protease inhibitor, is involved in the development of obesity, insulin resistance, energy metabolism, and reproduction. Its expression and regulation were studied in the human and rat placenta; however, the role of this adipokine in placental endocrine function has never been studied. The present study aimed to investigate the in vitro effects of vaspin on the endocrine function of the human placental syncytiotrophoblasts BeWo cell line and villous explants collected during the third trimester of pregnancy. BeWo cells (n=4) or villous explants (n=3) were cultured with vaspin at doses of 0.1, 1, and 10 ng/ml for 24, 48, and 72 h. The levels of progesterone (P4), estradiol (E2), human chorionic gonadotropin (hCG), and human placental lactogen (hPL) were determined in the culture medium via enzyme-linked immunosorbent assay (ELISA). In addition, the mRNA and protein expression of 3β-hydroxysteroid dehydrogenase (HSD3B1/3βHSD), aromatase (CYP19A1/CYP19), CGB3/hCG, and CSH1/hPL were determined via real-time PCR and Western blotting, respectively. We analyzed the role of pharmacological inhibitors of extracellular signal-regulated kinase (ERK1/2) and protein kinase A (PKA) in the vaspin action on hormone secretion. We observed that vaspin has a modulatory effect on the secretion and expression of placental hormones in BeWo cells and placentas from physiological pregnancies. However, in most cases, the effect was inhibitory on the parameters examined. Moreover, we noted that PKA participates in reducing E₂ secretion, while ERK1/2 is involved in hCG level. These findings indicate that vaspin is a new regulator of human placental endocrine function.

Diabetes mellitus (DM) is a known risk factor of aortic stenosis (AS). Among pathogenetic mechanisms associated with DM, increased oxidative stress plays a key role in the valvular degeneration progression. This study aimed to evaluate whether DM coexistence with severe AS is associated with parallel enhancement of oxidative stress in both circulating blood and stenotic valves. We enrolled 70 severe AS patients scheduled for aortic valve replacement, including 40 (57%) with concomitant type 2 DM (AS-DM) and 30 AS patients without type 2 DM (AS-nonDM). We assessed circulating oxidized phospholipids (oxPL), myeloperoxidase (MPO), advanced glycation end products (AGEs), and their soluble receptors (sRAGE). Valvular expression of oxPL and AGEs was determined using immunostaining of aortic valve samples. In the whole group of AS patients (n=70) oxPL, MPO and AGEs levels correlated with HbA1c concentration (r=0.79, p<0.001; r=0.60, p<0.001 and r=0.85, p<0.001, respectively). Similarly, oxPL and MPO were associated with plasma AGEs (r=0.66, p<0.001 and r=0.53, p<0.001). Moreover, serum MPO and oxPL correlated with aortic valve area (AVA; r= -0.49, p<0.001 and r= -0.45, p<0.001). Among 40 AS-DM patients we observed 2.55-fold higher serum oxPL, 1.38-fold higher serum MPO, 2.07-fold higher plasma AGEs and 2.22-fold higher plasma sRAGE compared to AS-nonDM (all p<0.001). In AS-DM group, valvular oxPL and AGEs expression was 1.8-fold and 1.6-fold higher when compared to AS-nonDM patients (all p<0.001). Of major importance, valvular expression of oxPL correlated both with serum oxPL and MPO levels (r=0.72 and r=0.63, respectively; both p<0.001). This study is the first to show that in severe AS patients with concomitant DM, valvular expression of oxPL is markedly higher compared to non-DM patients and associates with their circulating levels. Moreover, enhanced valvular oxidative stress was linked with glycemic status and more severe AS.