Effect of follicular fluid-derived exosomal mi RNAs on follicular dysplasia mediated via glycolysis of granulosa cells in patients with polycystic ovary syndrome and its mechanism / 中国生物制品学杂志

@#Objective To evaluate the effect of follicular fluid(FF)exosomal miRNAs on follicular dysplasia in patients with polycystic ovary syndrome(PCOS)mediated by glycolysis pathway of granulosa cells(GCs),and to explore the mechanism. Methods Three PCOS infertile patients and three non-PCOS infertile patients were recruited. The baseline hormone levels of the two groups were measured before ovulation induction. The bilateral FF was obtained by puncture after short-acting and long-term ovulation induction,and the exosomes were collected by ultracentrifugation and identified by transmission electron microscopy. The total exosomal RNA was extracted by Trizol method to construct the library,which was compared to the reference genome GRCh38 for statistical analysis after miRNA sequencing and quality control processing. Clustering Profiler R package was used to implement GO annotation analysis and KEGG pathway analysis of the differentially expressed genes(DEGs),and Omnipath software for miRNAs interaction analysis. A total of 16 miRNA were randomly selected and detected by qPCR to verify the accuracy of the miRNA sequencing results. Results Compared with the non-PCOS group,luteinizing hormone(LH),anti-Muerian hormone(AMH),testosterone and antral follicle counts in PCOS group increased significantly(t = 2. 479 ～ 9. 163,each P < 0. 05). The exosomes of FF in both groups showed the cup-shaped vesicles with clear edge and light staining in the center,with the diameters of 100 — 150 nm and intact structure,and the concentration was about 8 × 1010particles/mL. A total of 928 miRNAs were detected by miRNA sequencing. Compared with the non-PCOS group,59 differentially expressed miRNA(DEmiRNA)were screened out in exosomes of POCS group,of which 31 were up-regulated and 28 were down-regulated. The differential trend of gene expression detected by qPCR was highly similar to that of miRNA sequencing. In FF exosomes of PCOS patients,the glycolysis efficiency and apoptosis of GCs were significantly changed by miRNA regulating mRNA. PKM,PFKL and HK2 were the key target genes for miRNA to regulate GCs glycolysis,and SLC2A1 was the key target gene for miRNA to regulate GCs apoptosis. Conclusion The miRNAs in FF exosomes of PCOS patients can weaken the glycolysis of GCs while accelerate the apoptosis,thus reducing the production of ATP and lactic acid,resulting in follicular dysplasia.

The role of glucose metabolism reprogramming and its targeted therapeutic agents in inflammation-related diseases / 药学学报

Cells undergo glucose metabolism reprogramming under the influence of the inflammatory microenvironment, changing their primary mode of energy supply from oxidative phosphorylation to aerobic glycolysis. This process is involved in all stages of inflammation-related diseases development. Glucose metabolism reprogramming not only changes the metabolic pattern of individual cells, but also disrupts the metabolic homeostasis of the body microenvironment, which further promotes aerobic glycolysis and provides favourable conditions for the malignant progression of inflammation-related diseases. The metabolic enzymes, transporter proteins, and metabolites of aerobic glycolysis are all key signalling molecules, and drugs can inhibit aerobic glycolysis by targeting these specific key molecules to exert therapeutic effects. This paper reviews the impact of glucose metabolism reprogramming on the development of inflammation-related diseases such as inflammation-related tumours, rheumatoid arthritis and Alzheimer's disease, and the therapeutic effects of drugs targeting glucose metabolism reprogramming on these diseases.

Proliferation and metabolic patterns of smooth muscle cells during construction of tissue-engineered blood vessels / 中国组织工程研究

BACKGROUND:Seed cells are seeded on three-dimensional scaffold materials,and three-dimensional culture in bioreactors is a common in vitro tissue engineering culture method,but the changes in cell proliferation and metabolic patterns in bioengineered blood vessel construction are still unclear. OBJECTIVE:To explore the metabolic changes of cells such as oxygen consumption and their causes during the whole process of biological vascular tissue construction by in vitro bioreactor. METHODS:The self-built vascular bioreactor system was used as the platform;bovine vascular smooth muscle cells were used as the seed cells,and a conventional CO2 incubator provided the external gas environment for the cultivation process.Seed cells were seeded on a tubular porous polyglycolic acid scaffold material for three-dimensional culture,and the whole process included a one-week resting period and a seven-week pulsating tensile stress stimulation loading period.A non-invasive monitoring system was built,and the optical dissolved oxygen patch method was used to monitor the changes of dissolved oxygen in the culture solution in the reactor,and the glucose consumption and lactic acid production were measured by regular sampling.CCK-8 assay was used to determine the proliferation of smooth muscle cells on polyglycolic acid three-dimensional scaffold materials.Nicotinamide adenine dinucleotide oxidation state and reduction state ratio(NAD+/NADH)was utilized to understand cell proliferation and metabolism in the early stage of culture.RT-qPCR and western blot assay were applied to detect the expression of proliferation-related genes(Ki67)and glycolysis-related genes(GLUT-1,LDHA). RESULTS AND CONCLUSION:(1)The dissolved oxygen level in the culture solution was(4.314±0.380)mg/L from the cell injection to the end of the resting period(the first week),and gradually stabilized at(1.960±0.866)mg/L after the tensile stress stimulation(the last seven weeks);the two had significant changes(P<0.05).(2)The ratio of glucose consumption to lactic acid production in the cell culture medium YL/G increased rapidly after the cells were injected,and the highest value was above 1 on the fifth day,and then slow down to 0.5(The mean value of YL/G in the resting period was 0.89 and the mean value in the pressurized period was 0.57,P<0.05).(3)CCK-8 assay results showed that A450 value gradually increased after the cells were injected,and reached the highest value on the fifth day,reaching 3.17,and then slowly decreased.At the same time,it was found that Ki67 mRNA was up-regulated on the third day of culture,and then declined.The expression level of Ki67 protein was higher from the third day to the fifth day.(4)The detection of NAD+/NADH showed that the increase was obvious from the fifth to the seventh day after the injection of cells,and the expression of glycolysis-related genes(GLUT-1 and LDHA)was up-regulated and changed synchronously,and the relative expression was higher in the first five days.(5)The results showed that the tissue-engineered blood vessels were constructed using the vascular bioreactor and the smooth muscle cells in the early stage mainly proliferated and exhibited a metabolic feature of low oxygen consumption.The metabolic characteristics of high oxygen consumption were observed during the pulsatile tensile stress stimulation stage.

WIN55212-2 inhibits glycolysis and attenuates acute lung injury in septic mice by regulating mTOR/HIF-1α/PFKFB3 signaling pathway / 中国病理生理杂志

AIM:To investigate the effects of cannabinoid receptor agonist WIN55212-2(WIN)on acute lung injury(ALI)in septic mice,and to explore its potential mechanisms through glycolysis.METHODS:A mouse model of septic ALI was established by intraperitoneal injections of lipopolysaccharide(LPS).Male C57BL/6J mice were randomly divided into 4 groups(n=6):(1)control group;(2)LPS group,receiving intraperitoneal injections of LPS at 10 mg/kg;(3)LPS+WIN group,receiving 1 mg/kg WIN intraperitoneally 30 min prior to LPS injection;(4)LPS+WIN+MHY1485[mammalian target of rapamycin(mTOR)activator]group,receiving 10 mg/kg MHY1485 intraperitoneally 1 d before LPS injection and 1 mg/kg WIN plus 10 mg/kg MHY1485 30 min before LPS injection.Tissues were collected 24 h after modeling for analysis.Lung indexes were calculated,and histopathological changes of lung tissues were observed via he-matoxylin-eosin(HE)staining.Inflammatory cytokines interleukin-1β(IL-1β)and IL-10 in lung tissues,and lactic acid and lactate dehydrogenase A(LDHA)in serum were quantified using ELISA.The levels of mTOR/hypoxia-inducible fac-tor-1α(HIF-1α)/6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3(PFKFB3)signaling pathway-related proteins were assessed by Western blot.RESULTS:Compared with control group,the LPS group exhibited an increased lung in-dex,significant lung tissue damage,an increase in IL-1β levels(P＜0.05),a decrease in IL-10 levels(P＜0.05),and el-evated expressions of lactate and LDHA(P＜0.05),along with increased levels of phosphorylated mTOR(p-mTOR),HIF-1α and PFKFB3 proteins(P＜0.05).The LPS+WIN group showed improvements with a reduced lung index(P＜0.05),lessened lung injury,decreased IL-1β levels(P＜0.05),increased IL-10 levels(P＜0.05),and lower levels of lactic acid,LDHA,p-mTOR,HIF-1α,and PFKFB3(P＜0.05).Conversely,the LPS+WIN+MHY1485 group displayed increased lung indexes and lung tissue damage,elevated IL-1β levels(P＜0.05),reduced IL-10 levels(P＜0.05),and higher expressions of lactic acid,LDHA,p-mTOR,HIF-1α and PFKFB3(P＜0.05)compared to the LPS+WIN group.CONCLUSION:WIN55212-2 mitigates sepsis-induced ALI,potentially by modulating the mTOR/HIF-1α/PFKFB3 sig-naling pathway,thereby inhibiting glycolysis and alleviating inflammatory responses.

Effect of tumor-associated macrophage exosomes on glycolysis of pancreatic cancer cells by regulating KRAS signal pathway / 局解手术学杂志

Objective To investigate the effect of tumor-associated macrophage exosomes on glycolysis of pancreatic cancer cells and its mechanism.Methods The THP-1 cells were induced to differentiate into the M0 and M2 macrophages,and the exosomes(M0 exo and M2 exo)were extracted.The pancreatic cancer cells CAPAN-2 and ASPC-1 were divided into the PBS group,the M0 exo group,the M2 exo group and the M2 exo+siKRAS group,and co-incubated with equal volumes of PBS,10 μg/mL of M0 exo,10 μg/mL of M2 exo,and transfection of KRAS siRNA and 10 μg/mL of M2 exo,respectively.Transmission electron microscopy was used to observe the structure of exosomes;CCK-8 was used to detect the cell proliferation capacity;the kit was used to detect the glucose uptake rate and production level of lactic acid,and Western blot was used to detect the exosome markers expression,KRAS protein expression and ERK1/2 phosphorylation level.Results THP-1 was induced to differentiate into M2 macrophages expressing Arg-1 and IL-10 marker proteins.M0 exo and M2 exo had a bilayer membrane structure with a particle size of about 100 nm and expressed exosomal marker proteins of CD9,CD81,and TSG101.Compared with the PBS group,the cell proliferation,glucose uptake rate,production level of lactic acid of CAPAN-2 and ASPC-1 cells in the M2 exo group increased significantly(P<0.05),and the KRAS expression and ERK1/2 phosphorylation level were significantly increased(P<0.001).Compared with the M2 exo group,the proliferation,glucose uptake rate and production level of lactic acid of CAPAN-2 and ASPC-1 cells in the M2 exo+siKRAS group decreased significantly(P<0.05).Conclusion Tumor-associated macrophage exosomes can promote the glycolysis of pancreatic cancer cells via the activation of KRAS signaling pathway.

Research progress of glycolysis induced islet β cell dedifferentiation based on HIF-1α/PFKFB3 signaling pathway / 中国糖尿病杂志

Islet β cell dedifferentiation is one of the important reasons leading to insulin secretion defect or insulin resistance in patients with type 2 diabetes mellitus(T2DM).HIF-1α/PFKFB3 signaling pathway is a newly discovered biological pathway related to T2DM,which is involved in the induction of islet β cells dedifferentiation by anaerobic glycolysis under high glucose environment.This article reviews the research progress of the role of HIF-1α/PFKFB3 signaling pathway in glycolysis induced islet β cell dedifferentiation.

Changes in renal cell glycolysis and amino acid metabolism during cisplatin⁃induced acute kidney injury / 安徽医科大学学报

Objective @#To evaluate the change of energy metabolism during cisplatin ⁃induced acute kidney injury.@*Methods @#Adult CD⁃1 male mice were intraperitoneally inj ected with a single dose of cisplatin (20 mg/kg) , and renal function and renal tissue pathology were tested; gene expression was analyzed and signaling pathways were enriched in cisplatin ⁃treated renal tubular epithelial cells using transcriptome; the contents of renal glycolysis and amino acid metabolites were analyzed using liquid chromatography⁃tandem mass spectrometry ( LC⁃MS/MS) . @*Results@#Serum urea nitrogen and blood creatinine significantly increased in cisplatin ⁃treated mice. Pathological histology ob served swelling and shedding of renal tubular epithelial cells. Transcriptome analysis revealed that 2 632 genes were upregulated and 2 799 genes were downregulated in cisplatin ⁃treated HK⁃2 cells. GO and KEGG analyatin caused an upregulation of the oxidative phosphorylation pathway and a downregulation of the glycolysis pathway in renal tubular epithelial cells , further KEGG analysis demonstrated that cisplatin caused changes in the expression of amino acid genes in renal cells. Metabolomics showed that the contents of glycolytic intermediates and several amino acids were altered in the kidney of cisplatin ⁃treated mice. @*Conclusion @#Cisplatin ⁃induced acute renal injury is accompanied by modification in renal tubular cell glycolysis and amino acid metabolism.

Targeting STAT3 alleviates peritoneal fibrosis by regulating glycolysis and mesothelial-mesenchymal transition / 安徽医科大学学报

Objective @#To study the effect and mechanism of high glucose on mesothelial-mesenchymal transition (MMT) of peritoneal mesothelial cells (HMrSV5) , and the protective effect of pharmacological blocking of signal transducer and activator of transcription 3 (STAT3) on rat peritoneal fibrosis (PF) model . @*Methods @#The animals were divided into three groups : the sham group , the model group , and the STAT3 inhibitor group . A miniature per- itoneal dialysis catheter was implanted under the dorsal skin of rat and the rat peritoneal fibrosis model was induced by daily injection of high glucose dialysate . After 10 weeks , HE staining was used to evaluate the histology of the peritoneum , and the level of transforming growth factor-β1 (TGF-β1) in the peritoneum was measured by immuno- histochemistry . HMrSV5 was cultured in high glucose and the optimal stimulation concentration of high glucose was determined by Western blot. High glucose was used to stimulate HMrSV5 after successful transfection with si - STAT3 and Western blot was used to measure the protein level of STAT3 , p-STAT3 , and the key enzymes of glycol- ysis 6-phosphofructo-2-kinase/fructose-2 , 6-biphosphatase 3 (PFKFB3) and lactate dehydrogenase A (LDHA) .@*Results @#HE staining showed that administration of STAT3 inhibitor ( BP-1-102) could inhibit the thickening of subperitoneal tissue and the proliferation of vessels in HG dialysis rats . The expression of TGF-β1 in the rats perito- neum of the model group was significantly higher than that in the sham group , and the level of TGF-β1 was marked- ly lower in the STAT3 inhibitor group compared to the model group (P < 0. 05) . Compared to the control group , high glucose induced the up-regulation of α-smooth muscle actin ( α-SMA) , the down-regulation of E-cadherin and STAT3 activation in HMrSV5 (P < 0. 05) . Mesothelial cells treated with high glucose also exhibited high expres- sion of the key enzymes of glycolysis ( PFKFB3 , LDHA) ( P < 0. 05) , and si-STAT3 can effectively inhibit the overexpression of PFKFB3 and LDHA induced by high glucose ( P < 0. 05) . @*Conclusion @#STAT3 is involved in high glucose-induced HMrSV5 hyperglycolysis and MMT , and targeting STAT3 alleviates peritoneal fibrosis and an- giogenesis during peritoneal dialysis treatment in rats .

Effect of modified citrus pectin on glycolysis of rabbit articular chondrocytes / 国际生物医学工程杂志

Objective:To investigate the effect of modified citrus pectin (MCP) on the glucose metabolism of rabbit articular chondrocytes.Methods:The third generation (P3) rabbit knee chondrocytes were extracted and cultured with 0 μg/ml (MCP0, control group) and 500 μg/ml of MCP (MCP500) for 3 days. Chondrocytes (P2-P7)were cultured continuously, and each generation of chondrocytes was treated with MCP0 and MCP500 medium for 3 days. Chondrocytes were treated with interleukin-1β (IL-1β) for 1 day and then treated with MCP0 and MCP500 medium for 3 days, respectively. Chondrocytes were treated with 2-deoxy-glucose (2DG) for 1 day and then treated with MCP0 and MCP500 medium for 3 days, respectively. After three days of culture, the proliferation of chondrocytes was calculated by CCK-8. Glucose uptake activity and lactate production of chondrocytes were measured by glucose and lactate detection kits. The synthesis of type Ⅱ collagen (COL2A1) in sequential chondrocytes was investigated by immunofluorescence staining. The gene expression of COL2A1, proteoglycan ( ACAN), SOX9, hypoxia-inducible factor-1α ( HIF-1α), glucose transporter-1 ( Glut-1), pyruvate kinase M2 ( PKM2), lactate dehydrogenase-A ( LDHA) and glucose transporter-1 ( Glut-3) were further detected by RT-qPCR. Results:Compared with the control group, MCP treatment could increase the glucose uptake activity and lactate production of chondrocytes, and enhance the gene expression ability of HIF-1α, Glut-1, PKM2 and ACAN. Besides, MCP treatment could stimulate chondrocyte proliferation, maintain chondrocyte phenotype, increase lactate production, and upregulate the expression of COL2A1, ACAN, SOX9, HIF-1α, Glut-1, PKM2 and LDHA. After the treatment with IL-1β, MCP treatment could increase glucose uptake activity and upregulate the expression of COL2A1, ACAN, HIF-1α and Glut-1. After treatment with 2DG, MCP treatment could increase glucose uptake activity and upregulate the expression of SOX9, HIF-1α, PKM2 and Glut-3 genes. Conclusions:MCP can enhance the glucose uptake capacity of chondrocytes and increase the level of chondrocyte glycolytic metabolism.

Mechanism of Anti-inflammatory Effects of Bupi Yichang Pills on Inhibiting Glycolytic Metabolic Pathway in Mice with Experimental Colitis / 中药新药与临床药理

Objective To investigate the anti-inflammatory effects of Bupi Yichang Pills on mice with experimental colitis and its potential mechanism of action.Methods Dextran sulfate sodium(DSS)was used to model the experimental colitis,and low-,medium-and high-doses of Bupi Yichang Pills(1.5,3.0,6.0 g·kg-1·d-1)and Mesalazine(300 mg·kg-1·d-1)were fed at the same time.Mice were observed for general behavior and weighed.Hematoxylin-eosin staining was used to observe the pathological injury of colonic tissues.qPCR and ELISA were used to detect the levels of inflammatory cytokines(TNF-α,IL-1β,IL-6,IL-10,IL-35 and TGF-β1),qPCR and Western Blot were used to detect the mRNA and protein levels of glucose transporters and glycolytic kinases.Results Low-,medium-and high-doses of Bupi Yichang Pills significantly down-regulated disease activity index in colitis mice(P＜0.05,P＜0.01).The body mass and colon length were significantly increased,while colon mass,colon mass index and unit colon mass index were significantly reduced(P＜0.05,P＜0.01),and ulcer formation and inflammatory cell infiltration in colonic tissue were significantly improved.In addition,medium-and high-doses of Bupi Yichang Pills significantly down-regulated the mRNA levels and concentrations of pro-inflammatory cytokines including TNF-α,IL-1β and IL-6(P＜0.01),while significantly up-regulated the mRNA levels and concentrations of anti-inflammatory cytokines such as IL-10,IL-35 and TGF-β1(P＜0.01).We further found that high-dose of Bupi Yichang Pills significantly down-regulated the mRNA and protein expressions of glucose transporters(Glut1,Glut2,Glut4)and glycolytic kinases(HK2,Aldolase A,PKM2)in colonic tissue(P＜0.01).Conclusions Bupi Yichang Pills effectively alleviates DSS-induced experimental colitis,and its specific mechanism of action is related to the improvement of glycolytic metabolic pathways and the regulation of inflammatory cytokine expression.

Study on the mechanism of Compound lizard powder reducing cisplatin resistance in gastric cancer by regulating glycolysis / 中国药房

OBJECTIVE To explore the mechanism of Compound lizard powder reducing cisplatin resistance in gastric cancer by regulating glycolytic activity based on phosphoinositide 3-kinase（PI3K）/protein kinase B（Akt） signaling pathway. METHODS Human gastric cancer MKN45 and MKN45/DDP （cisplatin-resistant） cells were cultured in vitro and intervened with different mass concentrations of cisplatin （0.1， 0.2， 0.4， 0.8， 1.6， 3.2 μg/mL） to detect the survival rate， half inhibitory concentration （IC50） and drug resistance index. MKN45/DDP cells were inoculated subcutaneously in the right anterior axilla of nude mice to prepare a transplanted tumor model of gastric cancer. After successful modeling， they were randomly divided into model group， cisplatin group （0.002 g/kg）， Compound lizard powder group （2.8 g/kg） and combination group （the same dose as each single drug group）， with 8 nude mice in each group. Each administration group was given relevant solution， twice a week （cisplatin， i.p.） or twice a day （Compound lizard powder， i. g.）， for 4 consecutive weeks. During the experiment， the body weight of nude mice was monitored， and tumor volume and inhibitory rate of tumor were calculated. The levels of inflammatory factors （tumor necrosis factor- α， interleukin-6） in tumor tissue， the mRNA and protein expressions of multidrug resistance-associated protein 1 （MRP1）， P-glycoprotein （P-gp）， glucose transporter-1 （GLUT1） and lactate dehydrogenase A （LDHA）， as well as the protein expressions of PI3K， phosphorylated PI3K （p-PI3K）， Akt， phosphorylated Akt （p-Akt）， hexokinase-2 （HK2） and pyruvate kinase M2 （PKM2） were all detected. RESULTS With the intervention of different concentrations of cisplatin， the survival rate of MKN45/DDP-resistant cells was significantly higher than that of MKN45 parent cells （P＜0.05）. IC50 value of MKN45/DDP and MKN45 cells were（1.052 0±0.221 9） and （0.372 1±0.238 0）μg/mL， and the drug resistant index was 2.827. Compared with the model group， cisplatin group， Compound lizard powder group and combination group all had certain inhibitory effects on the tumor growth in nude mice； the inhibitory rates of tumor increased significantly （P＜0.05）； the levels of inflammatory factors， the mRNA and protein expressions of MRP1， P-gp， GLUT1 and LDHA （except for cisplatin group）， the phosphorylation levels of PI3K and Akt protein （except for cisplatin group） as well as the protein expressions of HK2 and PKM2 were decreased significantly， while the combination group was significantly better than the cisplatin group （P＜0.05）. CONCLUSIONS Compound lizard powder may inhibit tumor growth in transplanted tumor model nude mice with gastric cancer-resistant cells by reducing the secretion of tumor-related inflammatory factors， inhibiting the expression of glycolysis， drug resistance-related proteins and genes， inhibiting the activation of the PI3K/Akt signaling pathway， thus having a certain effect of enhancing cisplatin efficacy and reversing drug resistance.

Effect of Licoflavone A on Proliferation and Glycolysis of Gastric Cancer Cells Under Hypoxic Conditions / 中国实验方剂学杂志

ObjectiveTo investigate the effects of licoflavone A on the proliferation and glycolysis of gastric cancer cells in the hypoxic environment. MethodHuman gastric cancer AGS cells were classified into five groups： Normoxia， hypoxia， and low-， medium-， and high-dose （25， 50， 100 μmol·L-1， respectively） licoflavone A. The cells in other groups except the normoxia group were cultured in the environment with 5% O2 for 48 h. The cell counting kit-8 （CCK-8） and colony formation assay were employed to examine the proliferation of AGS cells. Cell migration was detected by the scratch assay. The protein and mRNA levels of hypoxia-inducible factor 1-alpha （HIF-1α）， glucose transporter 1 （GLUT1）， lactate dehydrogenase A （LDHA）， pyruvate kinase M2 （PKM2）， and hexokinase Ⅱ （HK2） in AGS cells were measured by Western blotting and real-time quantitative polymerase chain reaction （Real-time PCR）， respectively. The corresponding kits were used to determine glucose uptake and HK activity. ResultThe CCK-8 results showed that compared with the hypoxia group， the high- and medium-dose licoflavone A groups showed decreased proliferation rate of AGS cells at the time point of 24 h （P<0.01） and all the licoflavone A groups demonstrated decreased proliferation rate at the time point of 48 h （P<0.01）. Compared with the normoxia group， the hypoxia group showed increased number of clone formation of AGS cells （P<0.01）， which was decreased after the treatment with licoflavone A at high， medium， and low doses （P<0.01）. Compared with the normoxia group， the hypoxia group showed increased migration of AGS cells （P<0.01）， which was attenuated by the high， medium， and low doses of licoflavone A （P<0.01）. Compared with the normoxia group， the hypoxia group showed up-regulated mRNA levels of GLUT1， LDHA， PKM2， and HK2 （P<0.05， P<0.01）. Compared with those in the hypoxia group， the mRNA levels of GLUT1， LDHA， PKM2， and HK2 in the high-dose licoflavone A group， GLUT1， LDHA， and HK2 in the medium-dose licoflavone A group， and HK2 in the low-dose licoflavone A group were down-regulated （P<0.05， P<0.01）. The protein levels of HIF-1α， GLUT1， LDHA， PKM2， and HK2 in the hypoxia group were higher than those in the normoxia group （P<0.05， P<0.01）. Compared with those in the hypoxia group， the protein levels of HIF-1α， GLUT1， LDHA， PKM2， and HK2 in the high-dose licoflavone A group and HK2 in the medium- and low-dose licoflavone A groups were down-regulated （P<0.05， P<0.01）. The glucose uptake and HK activity were elevated in the hypoxia group compared with those in the normoxia group （P<0.01）. Compared with the hypoxia group， high-dose licoflavone A decreased the glucose uptake and HK activity， and medium-dose licoflavone A decreased the HK activity （P<0.01）. ConclusionLicoflavone A inhibits the proliferation of AGS cells under hypoxic conditions by regulating glycolysis in gastric cancer.

Role of Aerobic Glycolysis in Breast Cancer and Traditional Chinese Medicine Intervention： A Review / 中国实验方剂学杂志

Breast cancer has become the malignant tumor with the highest incidence rate. Although the emergence of new drugs has prolonged the overall survival of breast cancer patients， it still possesses a high recurrence and metastasis rate due to tumor heterogeneity and drug resistance. Glucose is the main source of energy metabolism for breast cancer cells， and the glucose metabolism of breast cancer cells is significantly different from that of normal breast cells. The high energy demand and rapid growth of breast cancer cells make their demand for glucose much higher than that of normal cells. Moreover， even under aerobic conditions， the glycolytic effect of breast cancer cells will be significantly enhanced to meet the high energy metabolism demand of breast cancer cells. The main reason for the enhanced glycolytic effect of breast cancer cells is the enhanced activity of glycolysis-related enzymes and regulatory factors， including pyruvate kinase， hexokinase， phosphofructokinase， lactate dehydrogenase， and glucose transporter protein. The metabolism process of glycolysis in breast cancer cells can be regulated by interfering with the activity of these enzymes and regulatory factors， thus inhibiting the proliferation of breast cancer， promoting apoptosis， and reversing drug resistance， invasion， and metastasis. Traditional Chinese medicine （TCM） has a long history of treating breast cancer and has made significant achievements in the aspects of anti-recurrence， metastasis， and drug resistance. In recent years， more and more research related to the intervention of aerobic glycolysis in breast cancer by TCM monomers， single-flavored TCM， and compounds has been conducted and has made great achievements. In addition， a large number of in vivo and in vitro experiments have shown that aerobic glycolysis is an important potential target for the treatment of breast cancer by TCM， but there is a lack of a comprehensive review and summary. On this basis， this paper elaborated on the roles of key targets in aerobic glycolysis and breast cancer and summarized the relevant studies on the treatment of breast cancer by intervention of glycolysis with TCM， with a view to providing new ideas for further research.

Research on the role of resveratrol against breast cancer / 中国药房

Breast cancer （BC） ranks first in the incidence rate of female malignant tumor， the notable features of which include high invasive behavior， high malignant degree and poor prognosis. Resveratrol， a plant antioxidant， has been identified as a potential therapeutic agent for the occurrence and progress of BC. This article explores the mechanism of resveratrol intervention in BC by evaluating several in vitro and in vivo studies. It was found that resveratrol can weaken the proliferation and survival ability of BC cells， suppress their growth， metastasis， and invasion， and reverse their resistance to adriamycin by promoting cell apoptosis， regulating autophagy， inhibiting glycolysis and regulating the tumor microenvironment， expressions of matrix metalloproteinases， epithelial-mesenchymal transition and drug-resistant proteins， etc. The limited number of clinical trial studies on resveratrol， mainly focusing on prevention effect of it on breast cancer， may be one of the reasons that affect the comprehensive evaluation of the anti-cancer efficacy of resveratrol.

Research on the role of resveratrol against breast cancer / 中国药房

Breast cancer （BC） ranks first in the incidence rate of female malignant tumor， the notable features of which include high invasive behavior， high malignant degree and poor prognosis. Resveratrol， a plant antioxidant， has been identified as a potential therapeutic agent for the occurrence and progress of BC. This article explores the mechanism of resveratrol intervention in BC by evaluating several in vitro and in vivo studies. It was found that resveratrol can weaken the proliferation and survival ability of BC cells， suppress their growth， metastasis， and invasion， and reverse their resistance to adriamycin by promoting cell apoptosis， regulating autophagy， inhibiting glycolysis and regulating the tumor microenvironment， expressions of matrix metalloproteinases， epithelial-mesenchymal transition and drug-resistant proteins， etc. The limited number of clinical trial studies on resveratrol， mainly focusing on prevention effect of it on breast cancer， may be one of the reasons that affect the comprehensive evaluation of the anti-cancer efficacy of resveratrol.

Sulforaphane regulates macrophage glycolysis and inhibits the progression of diabetic nephropathy by modulating the ALOX5/NF-κB signaling pathway / 安徽医科大学学报

Objective @# To investigate the effects of sulforaphane (SFN) in regulating the macrophage glycolysis via the arachidonate 5-lipoxygenase (ALOX5) /nuclear factor kappa B (NF-κB) signaling pathway on the progression of diabetic nephropathy (DN) . @*Methods @#Bioinformatics analysis was used to identify the target genes of SFN in the treatment of DN . Human proximal tubular epithelial cell line (HK-2 cells) was induced with 30 mmol/L high glucose (HG) to create an in vitro model of DN . HK-2 cells were divided into the following groups : normal glucose (NG) group , HG group , HG + SFN (3 mmol/L) group , HG + ALOX5 group , HG + SFN (3 mmol/L) + ALOX5 group , HG-treated macrophages + HK-2 group , HG + SFN (3 mmol/L) treated macrophages s + HK-2 group , HG + ALOX5 transfection treated macrophages + HK-2 group , HG + SFN (3 mmol/L) + ALOX5 transfection treated macrophages + HK-2 group . CCK-8 assay was used to detect cell viability , Terminal deoxynucleotidyl transferase- mediated dUTP nick-end labeling (TUNEL) method was used to detect cell apoptosis; glucose and lactate levels in the cells were measured using assay kits; Western blot was performed to detect the expression of ALOX5 , NF-κB , and glycolysis-related proteins hexokinase-2 ( HK2 ) , pyruvate kinase M2 ( PKM2 ) , glucose transporter 1 (GLUT1) in each group . Diabetic nephropathy (DN) mouse models were established using streptozotocin (STZ) and treated with SFN (0. 5 mg/kg) . Various biochemical parameters were measured in the mice , and kidney tissue pathology was examined using H&E staining. Western blot was used to detect the expression of glycolysis-related proteins (HK2 , PKM2 , GLUT1) in kidney macrophages . @*Results @# Bioinformatics analysis revealed ALOX5 as the target gene of SFN in treating DN . Compared to the HG group , SFN treatment enhanced HK-2 cell viability and in- hibited apoptosis (P < 0. 05) ; concurrently , SFN treatment suppressed HG-induced macrophage glycolysis-related protein and attenuated macrophage-mediated HK-2 cellular injury ( P < 0. 05) . Western blot results showed that SFN inhibited the expression of ALOX5 and NF-κB ( P < 0. 05) . The mouse experiment results showed that SFN treatment improved kidney function and pathological changes in the kidney of DN mice , and inhibited the related protein expression of acrophage glycolysis in kidney tissue (P < 0. 05) . @*Conclusion @#SFN improves the progression of DN by inhibiting the expression of macrophage glycolysis-related protein through the ALOX5/NF-κB signaling pathway .

Mechanism of catalpol regulating Th17 cell differentiation by interfering PKM2/LDHA expression / 中国药房

OBJECTIVE To investigate the mechanism of catalpol affecting the differentiation of helper T cell 17 （Th17） by interfering the expressions of pyruvate kinase M2 （PKM2） and lactate dehydrogenase A （LDHA）. METHODS The naive CD4+ T cells were selected from the spleen of C57BL/6 mice， and were differentiated into Th17 cells by adding directional differentiation stimulants for 72 hours. At the same time， the cells were treated with 0 （directed control）， 20， 40 and 80 μg/mL catalpol. The flow cytometry was used to detect the proportion of Th17 cell differentiation in cells； the colorimetric method was adopted to detect the levels of pyruvate and lactate in cell culture supernatant； mRNA expressions of retinoid-related orphan nuclear receptor gamma t （RORγt）， PKM2 and LDHA were detected by qRT-PCR method； Western blot was used to detect the expression levels of PKM2， LDHA， signal transducer and activator of transcription 3 （STAT3）， and phosphorylated STAT3 （p-STAT3） proteins in cells. RESULTS Compared with the directed control group， after 72 hours of treatment with 20， 40， 80 μg/mL catalpol， the differentiation ratio of Th17 cells were decreased by 6.74%， 8.41%， 9.24%， and the levels of pyruvate and lactate in the cell culture supernatant， the mRNA expressions of PKM2， LDHA and RORγt as well as the protein expressions of PKM2 and LDHA and the phosphorylation of STAT3 were significantly reduced （P＜0.05）. CONCLUSIONS Catalpol can reduce the glycolysis level by down-regulating the expressions of PKM2 and LDHA， thereby inhibiting the differentiation of Th17 cells.

Effect of viral infection on host cell metabolism: a review / 生物工程学报

As specialized intracellular parasite, viruses have no ability to metabolize independently, so they completely depend on the metabolic mechanism of host cells. Viruses use the energy and precursors provided by the metabolic network of the host cells to drive their replication, assembly and release. Namely, viruses hijack the host cells metabolism to achieve their own replication and proliferation. In addition, viruses can also affect host cell metabolism by the expression of auxiliary metabolic genes (AMGs), affecting carbon, nitrogen, phosphorus, and sulfur cycles, and participate in microbial-driven biogeochemical cycling. This review summarizes the effect of viral infection on the host's core metabolic pathway from four aspects: cellular glucose metabolism, glutamine metabolism, fatty acid metabolism, and viral AMGs on host metabolism. It may facilitate in-depth understanding of virus-host interactions, and provide a theoretical basis for the treatment of viral diseases through metabolic intervention.

Targeting microRNA-125b inhibited the metastasis of Alisertib resistance cells through mediating p53 pathway / 中华肿瘤杂志

Objective: To clarify the mechanisms involvement in Alisertib-resistant colorectal cells and explore a potential target to overcome Alisertib-resistance. Methods: Drug-resistant colon cancer cell line (named as HCT-8-7T cells) was established and transplanted into immunodeficient mice. The metastasis in vivo were observed. Proliferation and migration of HCT-8-7T cells and their parental cells were assessed by colony formation and Transwell assay, respectively. Glycolytic capacity and glutamine metabolism of cells were analyzed by metabolism assays. The protein and mRNA levels of critical factors which are involved in mediating glycolysis and epithelial-mesenchymal transition (EMT) were examined by western blot and reverse transcription-quantitative real-time polymerase chain reaction(RT-qPCR), respectively. Results: In comparison with the mice transplanted with HCT-8 cells, which were survival with limited metastatic tumor cells in organs, aggressive metastases were observed in liver, lung, kidney and ovary of HCT-8-7T transplanted mice (P<0.05). The levels of ATP [(0.10±0.01) mmol/L], glycolysis [(81.77±8.21) mpH/min] and the capacity of glycolysis [(55.50±3.48) mpH/min] in HCT-8-7T cells were higher than those of HCT-8 cells [(0.04±0.01) mmol/L, (27.77±2.55) mpH/min and(14.00±1.19) mpH/min, respectively, P<0.05]. Meanwhile, the levels of p53 protein and mRNA in HCT-8-7T cells were potently decreased as compared to that in HCT-8 cells (P<0.05). However, the level of miRNA-125b (2.21±0.12) in HCT-8-7T cells was significantly elevated as compared to that in HCT-8 cells (1.00±0.00, P<0.001). In HCT-8-7T cells, forced-expression of p53 reduced the colon number (162.00±24.00) and the migration [(18.53±5.67)%] as compared with those in cells transfected with control vector [274.70±40.50 and (100.00±29.06)%, P<0.05, respectively]. Similarly, miR-125b mimic decreased the glycolysis [(25.28±9.51) mpH/min] in HCT-8-7T cells as compared with that [(54.38±12.70)mpH/min, P=0.003] in HCT-8-7T cells transfected with control. Meanwhile, in comparison with control transfected HCT-8-7T cells, miR-125b mimic also significantly led to an increase in the levels of p53 and β-catenin, in parallel with a decrease in the levels of PFK1 and HK1 in HCT-8-7T cells (P<0.05). Conclusions: Silencing of p53 by miR-125b could be one of the mechanisms that contributes to Alisertib resistance. Targeting miR-125b could be a strategy to overcome Alisertib resistance.

Aerobic glycolysis in colon cancer is repressed by naringin via the HIF1Α pathway / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Metabolic reprogramming is a common phenomenon in cancer, with aerobic glycolysis being one of its important characteristics. Hypoxia-inducible factor-1α (HIF1Α) is thought to play an important role in aerobic glycolysis. Meanwhile, naringin is a natural flavanone glycoside derived from grapefruits and many other citrus fruits. In this work, we identified glycolytic genes related to HIF1Α by analyzing the colon cancer database. The analysis of extracellular acidification rate and cell function verified the regulatory effects of HIF1Α overexpression on glycolysis, and the proliferation and migration of colon cancer cells. Moreover, naringin was used as an inhibitor of colon cancer cells to illustrate its effect on HIF1Α function. The results showed that the HIF1Α and enolase 2 (ENO2) levels in colon cancer tissues were highly correlated, and their high expression indicated a poor prognosis for colon cancer patients. Mechanistically, HIF1Α directly binds to the DNA promoter region and upregulates the transcription of ENO2; ectopic expression of ENO2 increased aerobic glycolysis in colon cancer cells. Most importantly, we found that the appropriate concentration of naringin inhibited the transcriptional activity of HIF1Α, which in turn decreased aerobic glycolysis in colon cancer cells. Generally, naringin reduces glycolysis in colon cancer cells by reducing the transcriptional activity of HIF1Α and the proliferation and invasion of colon cancer cells. This study helps to elucidate the relationship between colon cancer progression and glucose metabolism, and demonstrates the efficacy of naringin in the treatment of colon cancer.