Resveratrol promotes autophagy through zinc ion and induces cardiomyocyte protection / 中国药理学通报

Aim To investigate whether resveratrol (Resveratrol, Res) induces cardiomyocyte protection by increasing intracellular zinc ion and its possible signal mechanism. Methods H9c2 cells were routinely cultured and 2-deoxyglucose (2-DG) was used to establish an endoplasmic reticulum stress (ERS) model. The experiment was randomly divided into control group, 2-DG group, Res +2-DG group, TPEN + Res + 2-DG group and 3-MA + Res +2-DG group. Cell viability was detected by MTT and CCK-8; the expression levels of ERS molecular chaperone proteins glucose-regulated protein 78 (GRP78), glucose-regulated protein 94 (GRP94) and autophagy proteins LC3 II / I, p62 and p-AMPK were detected by Western blot; the expression of LC3 protein was measured by cellular immunofluorescence; the mitochondrial membrane potential (Aijjm) and the intracellular zinc ion level were measured by laser scanning confocal microscope. Results Compared with the control group, 2-DG reduced cell activity and resveratrol inhibited the changes caused by 2-DG, which was reversed by zinc chelator TPEN. 2-DG increased GRP78 and GRP94 expression and resveratrol inhibited the protein changes caused by 2-DG, which was reversed by TPEN. 2-DG increased the expression of LC3 II / I, p-AMPK and decreased the expression of p62, and resveratrol promoted the effect of 2-DG. 2-DG increased the fluorescence intensity of LC3, and resveratrol enhanced the effect of 2-DG, which was reversed by TPEN and 3-MA. 2-DG reduced the red fluorescence intensity of mitochondrial TMRE and green fluorescence intensity of intracellular zinc ions, and resveratrol inhibited these changes caused by 2-DG, which was also reversed by TPEN and 3-MA. The above differences were all statistically significant (P < 0. 05). Conclusion Resveratrol increases intracellular zinc to promote ERS-induced autophagy and prevent the mPTP opening in H9c2 cardiac cells.

Hexokinase inhibitor 2-deoxyglucose combined with phenformin induces cell apoptosis of triple-negative breast cancer / 药学实践杂志

Objective To investigate the effect of phenformin combined with hexokinase inhibitor 2-deoxyglucose (2-DG) on the treatment of triple-negative breast cancer cell lines 4T1 and MDA-MB-231. Methods Following treatment with phenformin, 2-DG or phenformin combined with 2-DG on 4T1 and MDA-MB-231 cells for 48 h, the cell proliferation in each group was detected by SRB and the apoptosis of cells was detected by flow cytometry. The concentration of glucose and lactic acid in cell culture supernatant was detected by ELISA. The activity of mitochondrial respiratory chain complex Ⅰ was detected by FlexStation3 and the mitochondrial oxygen consumption (OCR) was assayed with the Seahorse X Fe Analyzer. Results The hexokinase expression (4.6±0.17,3.73±0.21), glucose consumption (356±31，397±42) μg/105 cells , Lactic acid concentration (5.59±0.52, 7.83±0.78) μmol/L in the supernatant of 4T1 and MDA-MB-231 cells in Phenformin group were higher than that in control group ( 1±0.15，1±0.12 ) , ( 289±25，301±32) μg/105cells , ( 2.37±0.18，4.01±0.45) μmol/L (P < 0.01). Even if the dose was reduced by 90%, the cell viability of phenformin combined with 2-DG group (64.63±2.28, 51.97±2.29) % was still higher than that of phenformin group (86.70±1.83, 85.53±1.46) % (P<0.001). The combination of the two drugs significantly promoted the apoptosis of 4T1 and MDA-MB-231. In addition, compared with the phenformin group (5.59±0.52, 7.83±0.78) μmol/L, the phenformin combined with 2-DG group (3.46±0.37, 5.18±0.62) μmol/L cell lactic acid production also greatly reduced (P<0.01). Compared with the phenformin or 2-DG single-drug group, the phenformin combined with 2-DG group can significantly inhibit the growth rate of tumors in tumor-bearing mice (P<0.01). The median survival time of tumor-bearing mice in the phenformin combined with 2-DG group was 72.5 d, which was higher than that in the phenformin group 57 d and 2-DG group 55.5 d (P<0.01). Conclusion Hexokinase inhibitor 2-DG significantly enhances the therapeutic effects of phenformin on triple-negative breast cancer cells.

Primary Malignant Melanoma of the esophagus in a 39-year-old Filipino male: A case report

@#Primary malignant melanoma of the esophagus (PMME) is an exceptionally rare condition, representing a mere 0.1 to 0.2% of esophageal cancers, and accounting for just 0.1 to 0.5% of all melanomas. This case involves a 39 -year-old Filipino male who sought medical attention after an episode of choking. Subsequently, endoscopy with biopsy revealed a mass in the distal third of the esophagus, ultimately diagnosed as PMME based on histopathology and immunohistochemistry. FDG-PET/CT scan revealed a hypermetabolic distal esophageal mass and a confluent upper paratracheal lymphadenopathy. He was initially treated with Pembrolizumab, Nivolumab, and Ipilimumab immunotherapy. However, post-treatment FDG PET/CT scans unveiled metabolic progression of the esophageal mass with new hypermetabolic cervical lymph nodes, necessitating a shift to carboplatin and paclitaxel chemotherapy. After two cycles, there was a notable metabolic regression of the mass and paratracheal node with metabolic resolution of the cervical lymph node. An additional 2 cycles of chemotherapy were given, aimed to further reduce the size of the tumor, however, a succeeding follow-up study revealed metabolic progression of the mass. Surgical resection of both the esophageal mass and paratracheal nodes became imperative. The aggressive characteristics, metastasis at early diagnosis, and lack of effective treatment have contributed to the poor prognosis of PMME. Total esophagectomy is the preferred method of treatment. Chemotherapy and immunotherapy may be used in advanced diseases but with variable efficacy. The utilization of FDG PET/CT scans plays a crucial role in both the initial staging and the ongoing assessment of treatment response in patients diagnosed with PMME. This advanced imaging modality offers valuable insights into the extent of the disease and aids clinicians in evaluating the effectiveness of the chosen therapeutic interventions. Given the rarity and challenges associated with PMME, a multidisciplinary approach integrating surgical, medical, and imaging strategies is essential for comprehensive patient care.

Targeting glutamine utilization to block metabolic adaptation of tumor cells under the stress of carboxyamidotriazole-induced nutrients unavailability

Tumor cells have unique metabolic programming that is biologically distinct from that of corresponding normal cells. Resetting tumor metabolic programming is a promising strategy to ameliorate drug resistance and improve the tumor microenvironment. Here, we show that carboxyamidotriazole (CAI), an anticancer drug, can function as a metabolic modulator that decreases glucose and lipid metabolism and increases the dependency of colon cancer cells on glutamine metabolism. CAI suppressed glucose and lipid metabolism utilization, causing inhibition of mitochondrial respiratory chain complex I, thus producing reactive oxygen species (ROS). In parallel, activation of the aryl hydrocarbon receptor (AhR) increased glutamine uptake via the transporter SLC1A5, which could activate the ROS-scavenging enzyme glutathione peroxidase. As a result, combined use of inhibitors of GLS/GDH1, CAI could effectively restrict colorectal cancer (CRC) energy metabolism. These data illuminate a new antitumor mechanism of CAI, suggesting a new strategy for CRC metabolic reprogramming treatment.

Bioengineered miR-328-3p modulates GLUT1-mediated glucose uptake and metabolism to exert synergistic antiproliferative effects with chemotherapeutics

MicroRNAs (miRNAs or miRs) are small noncoding RNAs derived from genome to control target gene expression. Recently we have developed a novel platform permitting high-yield production of bioengineered miRNA agents (BERA). This study is to produce and utilize novel fully-humanized BERA/miR-328-3p molecule (hBERA/miR-328) to delineate the role of miR-328-3p in controlling nutrient uptake essential for cell metabolism. We first demonstrated successful high-level expression of hBERA/miR-328 in bacteria and purification to high degree of homogeneity (>98%). Biologic miR-328-3p prodrug was selectively processed to miR-328-3p to suppress the growth of highly-proliferative human osteosarcoma (OS) cells. Besides glucose transporter protein type 1, gene symbol solute carrier family 2 member 1 (GLUT1/), we identified and verified large neutral amino acid transporter 1, gene symbol solute carrier family 7 member 5 (LAT1/) as a direct target for miR-328-3p. While reduction of LAT1 protein levels by miR-328-3p did not alter homeostasis of amino acids within OS cells, suppression of GLUT1 led to a significantly lower glucose uptake and decline in intracellular levels of glucose and glycolytic metabolite lactate. Moreover, combination treatment with hBERA/miR-328 and cisplatin or doxorubicin exerted a strong synergism in the inhibition of OS cell proliferation. These findings support the utility of novel bioengineered RNA molecules and establish an important role of miR-328-3p in the control of nutrient transport and homeostasis behind cancer metabolism.

Combination of a single-chain variable fragment JZC00 with 2-deoxyglucose inhibited tumor growth in murine models / 中国药科大学学报

@#This study aimed to investigate the antitumor efficacy of a single-chain variable fragment JZC00 combined with 2-deoxyglucose(2-DG)on murine non-small lung cancer cell and breast cancer cell models. JZC00 was expressed by E. coli and identified using SDS-PAGE and Western blot. The combination inhibited the proliferation of LLC and 4T1 cells. The concentration of glucose and lactic acid in the medium were determined by glucose and lactate kit, respectively, then calculated the tumor cell glucose uptake inhibition rate and lactate release inhibition rate. In vivo, the tumor volume and tumor weight were analyzed after 15-day treatment. The results showed that the molecular weight of JZC00 expressed was correct, and it could inhibit the proliferation of tumor cells in vitro. JZC00 and 2-DG could inhibit the glycolysis of tumor cells, respectively, and JZC00 combined with 2-DG could inhibit glycolysis synergistically. When hypoxic microenvironment was induced in vitro, the inhibition of glycolysis by JZC00 treatment decreased. However, it was reversed with the addition of 2-DG. The in vivo models the combination showed a significantly improved tumor suppressive effect compared with JZC00 treated group, suggesting that 2-DG could improve the anti-tumor effect of anti-angiogenic antibodies and its combination has the potentialial value in the treatment of solid tumors.

Silibinin ameliorates hepatic lipid accumulation and oxidative stress in mice with non-alcoholic steatohepatitis by regulating CFLAR-JNK pathway

Non-alcoholic steatohepatitis (NASH) is a chronic metabolic syndrome and the CFLAR-JNK pathway can reverse the process of NASH. Although silibinin is used for the treatment of NASH in clinical, its effect on CFLAR-JNK pathway in NASH remains unclear. This study aimed to investigate the effect of silibinin on CFLAR-JNK pathway in NASH models both and . The study was performed using male C57BL/6 mice fed with methionine- choline-deficient diet and simultaneously treated with silibinin for 6 weeks. The study was performed by using mouse NCTC-1469 cells which were respectively pretreated with oleic acid plus palmitic acid, and adenovirus-down for 24 h, then treated with silibinin for 24 h. After the drug treatment, the key indicators involved in CFLAR-JNK pathway including hepatic injury, lipid metabolism and oxidative stress were determined. Silibinin significantly activated CFLAR and inhibited the phosphorylation of JNK, up-regulated the mRNA expression of and , reduced the activities of serum ALT and AST and the contents of hepatic TG, TC and MDA, increased the expression of NRF2 and the activities of CAT, GSH-Px and HO-1, and decreased the activities and expression of CYP2E1 and CYP4A . These effects were confirmed by the experiments. Silibinin prevented NASH by regulating CFLAR-JNK pathway, and thereby on one hand promoting the -oxidation and efflux of fatty acids in liver to relieve lipid accumulation, and on the other hand inducing antioxidase activity (CAT, GSH-Px and HO-1) and inhibiting pro-oxidase activity (CYP2E1 and CYP4A) to relieve oxidative stress.

2-deoxyglucose inhibits angiogenesis of rheumatoid arthritis via activating AMPK pathway / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effects of 2-deoxyglucose inhibiting synovial pannus of adjuvant arthritis rats and to explore its potential mechanism of inhibiting angiogenesis by investigating proliferation, migration and matrigel tube formation assay .</p><p><b>METHODS</b>The effect of 2-DG on synovial pannus was evaluated by histopathology of HE staining; HUVEC proliferation was determined by CCK-8 method; migration of FLS were determined by transwell; matrigel tube formation assay was made for assessing tube number of HUVEC; p-AMPK and Bcl-2 were detected by Western blot assay; AMPK signaling pathway in HUVEC was inhibited by compound C, which is an inhibitor of AMPK activation.</p><p><b>RESULTS</b>2-DG (200 mg/kg) obviously decreased appearance of synovial pannus ( < 0.01); , 2-DG (0.5 mmol/L and/or 5 mmol/L) obviously inhibited proliferation, migration and tube number of HUVEC ( < 0.01 or < 0.001), and its effects on HUVEC were reversed by using AMPK antagonist (Compound C); Western blot showed that 2-DG (5 mmol/L) increased expression of p-AMPK and decreased expression of Bcl-2 ( < 0.05).</p><p><b>CONCLUSIONS</b>Activating AMPK pathway and decreasing expression of Bcl-2 may the potential mechanism by which 2-DG contributes to anti-angiogenesis and effects of inhibiting proliferation, migration and tube number of HUVEC.</p>

The diagnostic values of MRI, enhanced CT and PET-CT in detecting skull base invasion of nasopharyngeal carcinomas: A comparative study / 肿瘤

Objective: To compare the diagnostic values of magnetic resonance imaging (MRI), enhanced computed tomography (CT), and positron emission tomography (PET)-CT using 18F-fluoro-2-deoxyglucose (FDG) in detecting skull base invasion of nasopharyngeal carcinoma (NPC) and provide the evidence for differentiated diagnosis of the skull base invasion of NPC. Methods: The fifty seven patients were scanned by MRI, enhanced CT, and PET-CT. The three imaging examinations were finished within 20 days. The diagnosis standards were based on histopathologic findings or clinical and imaging follow-up results within 6 months. The sensitivity, specificity, accuracy, positive predictive value (PPV) and negative predictive value (NPV) of the three scanning technologies were compared. Results: For detecting skull base invasion of NPC, the sensitivity of enhanced CT, MRI, and PET-CT were 68.18%, 84.09%, and 97.67%, respectively; the specificity were 76.92%, 69.23%, and 57.14%, respectively; the accuracy were 70.18%, 80.70%, and 87.72%, respectively; PPV were 90.90%, 90.24%, and 87.50%, respectively; NPV were 41.67%, 56.25%, and 88.89%, respectively. PET-CT was better than enhanced CT in sensitivity, accuracy, and NPV (P <0.05). It was also better than MRI in sensitivity and NPV (P <0.05). Conclusion: Among three imaging technologies,PET-CT has obvious advantage in detecting skull base invasion of NPC patients, especially for new patients.

Effects of Insulin, 2-Deoxyglucose and Dichloroacetate on Acute Focal Cerebral Ischemia in Rats

Hyperglycemia has been reported to worsen the tolerance of the brain to ischemia, and it has therefore been recommended that patient undergoing neurosurgical procedures not receive glucose-containing solutions. Since ischemic events lead to increased lactate production and accumulation and hence neuronal damage, the present study was designed to test the effect of insulin-induced hypoglycemia and decreased lactate by 2-Deoxyglucose and Dichloroacetate on focal cerebral ischemia in rats. Although the pre and post-ischemic blood glucose levels of control group and Dichloroacetate group showed no change, the blood glucose level of 2-Deoxyglucose group showed a significant increase(p=0.001), and insulim group a significant decrease(p=0.004). The reducing effects on the infarct zone in these three treated groups were found with statistical significance. As compared with control group, the tissue lactate levels of treated groups were decreased in both infarct zone and border zone but these data did not show statistical significance. From these observations, it is suggested that reduction of lactate production and accumulation could be beneficial by affording neuronal protection in ischemic tissues.

THE LAMINARY PROJECTION OF ACUPUNCTURE AND NOXIOUS INFORMATION IN THE RAT SPINAL CORD DORSAL HORN / 西安交通大学学报(医学版)

The relative quantitative [~3H]-2-deoxyglucose (2DG)method was used to study the laminary projection of acupuncture and noxious information in the rat spinal cord dorsal horn. The noxious information projected to lamina Ⅰ、Ⅱ、Ⅳ、Ⅴ、Ⅵ and Ⅹ, and the acupuncture information projected to lamina Ⅰ、Ⅱ、Ⅳ and Ⅴ in the spinal cord dorsal horn. The noxious and acupuncture information had overlapping projection areas in the spinal cord. It is in such areas that the two types of information may be interacted on each other and consequently the nociception is modulated at the spinal level.