Mechanisms of Dangua Fang(丹瓜方)in multi-target and multi-method regulation of glycolipid metabolism based on phosphoproteomics

OBJECTIVE:To explore the mechanism of Dangua Fang(丹瓜方,DGR)in multi-target and multi-method regulation of glycolipid metabolism based on phosphoproteomics.METHODS:Sprague-Dawley rats with normal glucose levels were randomly divided into three groups,including a conventional diet control group(Group A),high-fat-highsugar diet model group(Group B),and DGR group(Group C,high-fat-high-sugar diet containing 20.5 g DGR).After 10 weeks of intervention,the fasting blood glucose(FBG),2 h blood glucose[PBG;using the oral glucose tolerance test(OGTT)],hemoglobin A1c(HbA1c),plasma total cholesterol(TC),and triglycerides(TG)were tested,and the livers of rats were removed to calculate the liver index.Then,hepatic portal TG were tested using the Gross permanent optimization-participatiory action planning enzymatic method and phosphoproteomics was performed using liquid chromatography with tandem mass spectrometry(LC-MS/MS)analysis followed by database search and bioinformatics analysis.Finally,cell experiments were used to verify the results of phosphoproteomics.Phosphorylated mitogen-activated protein kinase kinase kinase kinase 4(MAP4k4)and phosphorylated adducin 1(ADD1)were detected using western blotting.RESULTS:DGR effectively reduced PBG,TG,and the liver index(P<0.05),and significantly decreased HbA1c,TC,and hepatic portal TG(P<0.01),showed significant hematoxylin and eosin(HE)staining,red oil O staining,and Masson staining of liver tissue.The total spectrum was 805334,matched spectrum was 260471,accounting for accounting 32.3%,peptides were 19995,modified peptides were 14671,identified proteins were 4601,quantifiable proteins were 4417,identified sites were 15749,and quantified sites were 14659.Based on the threshold of expression fold change(>1.2),DGR upregulated the modification of 228 phosphorylation sites involving 204 corresponding function proteins,and downregulated the modification of 358 phosphorylation sites involving 358 corresponding function proteins,which included correcting 75 phosphorylation sites involving 64 corresponding function proteins relating to glycolipid metabolism.Therefore,DGR improved biological tissue processes,including information storage and processing,cellular processes and signaling,and metabolism.The metabolic functions regulated by DGR mainly include energy production and conversion,carbohydrate transport and metabolism,lipid transport and metabolism,inorganic ion transport and metabolism,secondary metabolite biosynthesis,transport,and catabolism.In vitro phosphorylation validation based on cell experiments showed that the change trends in the phosphorylation level of MAP4k4 and ADD1 were consistent with that of previous phosphoproteomics studies.CONCLUSION:DGR extensively corrects the modification of phosphorylation sites to improve corresponding glycolipid metabolism-related protein expression in rats with glycolipid metabolism disorders,thereby regulating glycolipid metabolism through a multi-target and multi-method process.

Modified Linggui Zhugan Decoction Ameliorates Glycolipid Metabolism and Inflammation via PI3K-Akt/m TOR-S6K1/AMPK-PGC-1α Signaling Pathways in Obese Type 2 Diabetic Rats

Objective: To investigate the protective effects of modified Linggui Zhugan Decoction(MLZD), a traditional Chinese medicine formula, on obese type 2 diabetes mellitus(T2 DM) rats. Methods:Fifty Sprague-Dawley rats were randomly divided into 5 groups by a random number table, including normal, obese T2 DM(ob-T2 DM), MLZD low-dose [MLDZ-L, 4.625 g/(kg·d)], MLZD middle-dose [MLD-M,9.25 g/(kg·d) ] and MLZD high-dose [MLD-H, 18.5 g/(kg·d)] groups,10 rats in each group. After 4-week intervention, blood samples and liver, pancreas, muscle tissues were collected to assess the insulin resistance(IR), blood lipid, adipokines and inflammation cytokines. The alteration of phosphatidylinositol 3 kinase(PI3 K)-protein kinase B(PKB or Akt)/the mammalian target of rapamycin(mTOR)-ribosome protein subunit 6 kinase 1(S6 K1)/AMP-activated protein kinase(AMPK)-peroxisome proliferator-activated receptor gamma coactivator 1 alpha(PGC-1α) pathways were also studied. Results: MLZD dose-dependently reduced fasting blood glucose,fasting insulin, homeostasis model of assessment for IR index and increased insulin sensitive index compared with ob-T2 DM rats(P<0.05). Similarly, total cholesterol, triglyceride, low-density lipoprotein cholesterol and free fatty acids were also decreased compared with ob-T2 DM rats after 4-week treatment(P<0.05 or P<0.01).Improvements in adipokines and inflammatory cytokines were observed with a raised level of adiponectin and a reduced level of leptin, resistin, tumor necrosis factor-α and interleukin-6(P<0.05 or P<0.01). MLZD regulated the PI3 K-Akt/mTOR-S6 K1/AMPK-PGC-1α pathways and restored the tissue structure of liver and pancreas(P<0.05 or P<0.01). Conclusions: MLZD ameliorated glycolipid metabolism and inflammation, which may be attributed to the regulation of PI3 K-Akt/mTOR-S6 K1/AMPK-PGC-1α pathways.

Functionalized gadofullerene ameliorates impaired glycolipid metabolism in type 2 diabetic mice

The soaring global prevalence of diabetes makes it urgent to explore new drugs with high efficacy and safety.Nanomaterial-derived bioactive agents are emerging as one of the most promising candidates for biomedical application.In the present study,we investigated the anti-diabetic effects of a functionalized gadofullerene(GF)using obese db/db and non-obese mouse model of type 2 diabete mellitus(MKR)mouse type 2 diabetes mellitus(T2DM)models.In both mouse models,the diabetic phenotypes,including hyperglycemia,impaired glucose tolerance,and insulin sensitivity,were ameliorated after two or four weeks of intraperitoneal administration of GF.GF lowered blood glucose levels in a dose-dependent manner.Importantly,the restored blood glucose levels could persist ten days after withdrawal of GF treatment.The hepatic AKT/GSK3β/FoxO1 pathway is shown to be the main target of GF for rebalancing gluconeogenesis and glycogen synthesis in vivo and in vitro.Furthermore,GF treatment significantly reduced weight gain of db/db mice with reduced hepatic fat storage by the inhibition of de novo lipogenesis through m TOR/S6K/SREBP1 pathway.Our data provide compelling evidence to support the promising application of GF for the treatment of T2DM.

Hypoglycemic mechanism of Tegillarca granosa polysaccharides on type 2 diabetic mice by altering gut microbiota and regulating the PI3K-akt signaling pathwaye

Type 2 diabetes mellitus(T2DM)is a complex metabolic disease threatening human health.We investigated the effects of Tegillarca granosa polysaccharide(TGP)and determined its potential mechanisms in a mouse model of T2DM established through a high-fat diet and streptozotocin.TGP(5.1×10^(3) Da)was composed of mannose,glucosamine,rhamnose,glucuronic acid,galactosamine,glucose,galactose,xylose,and fucose.It could significantly alleviate weight loss,reduce fasting blood glucose levels,reverse dyslipidemia,reduce liver damage from oxidative stress,and improve insulin sensitivity.RT-PCR and Western blotting indicated that TGP could activate the phosphatidylinositol-3-kinase/protein kinase B signaling pathway to regulate disorders in glucolipid metabolism and improve insulin resistance.TGP increased the abundance of Allobaculum,Akkermansia,and Bifidobacterium,restored the microbiota abundance in the intestinal tracts of mice with T2DM,and promoted short-chain fatty acid production.This study provides new insights into the antidiabetic effects of TGP and highlights its potential as a natural hypoglycemic nutraceutical.

Effect of individualized nutrition interventions on clinical outcomes of pregnant women with gestational diabetes mellitus

BACKGROUND Gestational diabetes mellitus(GDM)can lead to excessive pregnancy weight gain(PWG),abnormal glucolipid metabolism,and delayed lactation.Therefore,it is necessary to provide appropriate and effective interventions for pregnant women with GDM.AIM To clarify the effects of individualized nutrition interventions on PWG,glucolipid metabolism,and lactation in pregnant women with GDM.METHODS The study population consisted of 410 pregnant women with GDM who received treatment at the Northern Jiangsu People's Hospital of Jiangsu Provinceand Yangzhou Maternal and Child Health Hospital between December 2020 and December 2022,including 200 who received routine in-terventions[control(Con)group]and 210 who received individualized nutrition interventions[research(Res)group].Data on PWG,glucolipid metabolism[total cholesterol,(TC);triglycerides(TGs);fasting blood glucose(FPG);glycosylated hemoglobin(HbA1c)],lactation time,perinatal complications(cesarean section,premature rupture of membranes,postpartum hemorrhage,and pregnancy-induced hypertension),and neonatal adverse events(premature infants,fetal macrosomia,hypo-glycemia,and respiratory distress syndrome)were collected for comparative analysis.RESULTS The data revealed markedly lower PWG in the Res group vs the Con group,as well as markedly reduced TG,TC,FPG and HbA1c levels after the intervention that were lower than those in the Con group.In addition,obviously earlier lactation and statistically lower incidences of perinatal complications and neonatal adverse events were observed in the Res group.CONCLUSION Individualized nutrition interventions can reduce PWG in pregnant women with GDM,improve their glucolipid metabolism,and promote early lactation,which deserves clinical promotion.

Research progress of artesunate in diabetes and its complications

Diabetes is a metabolic disease characterized by abnormally elevated blood glucose levels.Persistent hyperglycemia leads to diabetic nephropathy,diabetic retinopathy,diabetes with periodontal disease and other diabetic complications.These diseases have become the main causes of disability and death in diabetic patients.Artesunate is well known as an antimalarial drug for controlling malaria symptoms.Current studies have shown that artesunate improves diabetes and its complications by protecting islet cells,improving glucose and lipid metabolism,anti-inflammatory and immune regulation.Based on the research status in recent years,this paper focuses on the mechanism of artesunate in diabetes and its complications,to provide a theoretical basis for future diabetes research.

Mechanisms of Dangua Recipe in Improving Glycolipid Metabolic Disorders Based on Transcriptomics

Objective:To explore the mechanisms of Dangua Recipe(DGR)in improving glycolipid metabolism based on transcriptomics.Methods:Sprague-Dawley rats with normal glucose level were divided into 3 groups according to a random number table,including a conventional diet group(Group A),a DGR group(Group B,high-calorie diet+20.5 g DGR),and a high-calorie fodder model group(Group C).After 12 weeks of intervention,the liver tissue of rats was taken.Gene sequence and transcriptional analysis were performed to identify the key genes related to glycolipid metabolism reflecting DGR efficacy,and then gene or protein validation of liver tissue were performed.Nicotinamide phosphoribosyl transferase(Nampt)and phosphoenolpyruvate carboxykinase(PEPCK)proteins in liver tissues were detected by enzyme linked immunosorbent assay,fatty acid synthase(FASN)protein was detected by Western blot,and fatty acid binding protein 5(FABP5)-mRNA was detected by quantitative real-time polymerase chain reaction.Furthermore,the functional verification was performed on the diabetic model rats by Nampt blocker(GEN-617)injected in vivo.Hemoglobin A1c(HbA1c),plasma total cholesterol and triglycerides were detected.Results:Totally,257 differentialdominant genes of Group A vs.Group C and 392 differential-dominant genes of Group B vs.Group C were found.Moreover,11 Gene Ontology molecular function terms and 7 Kyoto Encyclopedia of Genes and Genomes enrichment pathways owned by both Group A vs.Group C and Group C vs.Group B were confirmed.The liver tissue target validation showed that Nampt,FASN,PEPCK protein and FABP5-mRNA had the same changes consistent with transcriptome.The in vivo functional tests showed that GEN-617 increased body weight,HbA1c,triglyceride and total cholesterol levels in the diabetic rats(P<0.05 or P<0.01);while all the above-mentioned levels(except triglyceride)were decreased significantly by GEN-617 combined with DGR intervention(P<0.05 or P<0.01).Conclusion:Nampt activation was one of the mechanisms about DGR regulating glycolipid metabolism.

Stereoselective metabolic disruption of cypermethrin by remolding gut homeostasis in rat

Cypermethrin(CYP), a prototypical synthetic pyrethroid, reportedly causes metabolic disruption, while its stereoselective impact remains elusive. This study initially revealed that only α-CYP caused significant weight loss at 8.5 mg/(kg·day) in rats. All three CYP isomers caused the accumulation of hepatic glycogen, and hyperlipemia phenotype as the increment of total triglyceride. Rats treated with α-CYP had markedly high blood glucose levels and homeostasis model assessment of insulin resistance index. The systematic inflammation of θ-CYP group rats was evidenced by high lipopolysaccharide-binding protein levels and abnormalities of leukocytes indices. By examining the gut microbiome, we found thatα-CYP-treated rats had low contents of Firmicutes and high levels of Verrucomicrobia while Elusimicrobia was enriched in the β-CYP group. The increasing alpha diversity in the θ-CYP group may be due to the dominance of pathogenic bacteria and the increase of probiotics to counteract adverse effects. Exclusively, the α-CYP group enriched total short-chain fatty acids(SCFAs), whereas most SCFAs depleted in the θ-CYP group. The correlation analysis further found Firmicutes, an energy storage modulator, was positive to body weight(BW),while SCFAs exerted the opposite, confirming the low BW in α-CYP. Blood glucose that correlated well with SCFAs and Verrucomicrobia can be accounted for the discrepancy betweenα-CYP and θ-CYP. Overall, the three isomers exerted stereoselective glycolipid disruption in rats, and gut homeostasis acted as vital indicators.

Therapeutic Effect of Metformin on Patients with Polycystic Ovary Syndrome with Normal Insulin Sensitivity: A Retrospective Study

Objective:This study aimed at elucidating the therapeutic effectiveness of metformin in polycystic ovary syndrome(PCOS)patients with normal clinical indices of insulin sensitivity.Methods:This study was a retrospective cohort study.Based on their homeostasis model of assessment of insulin resistance(HOMA-IR)results,PCOS patients(n=113)were allocated to either the IR Group(IR,HOMA-IR>2.69)or the non-IR Group(NIR,HOMA-IR≤2.69).Metformin was administered to all patients,and their medical history,menstrual status,and endocrine and metabolic characteristics were obtained before and after treatment.The one-way analysis of variance was used to determine the differences between the IR and NIR groups.The paired t-test was used to determine the differences between pre-and posttreatment results.Results:Analyses of baseline characteristics showed that the incidence of IR and glycolipid metabolism is higher within elder PCOS patients.Metformin improved menstrual cycle and ovulation rate in NIR patients,similar to those in IR patients.In the NIR group,metformin decreased luteinizing hormone(LH)levels,free androgen index(FAI)levels,body mass index,and waist circumference,but had no obvious effect on glucose,insulin,and lipid profiles.Whereas in the IR group,metformin not only decreased LH and FAI levels,but also improved glycolipid metabolism.Conclusions:The effect of metformin on PCOS patients with normal insulin sensitivity index suggests that the mechanism of menstrual improvement may not be related to IR.In addition,metformin can improve glycolipid metabolism.Therefore,metformin is suitable for long-term treatment of PCOS and should be more widely used.

Efficacy of Huoxue Qianyang Qutan Recipe on essential hypertension:A randomized,double-blind,placebo-controlled trial

Background:Hypertension,a prevalent disease,is a significant risk factor for coronary heart disease.Huoxue Qianyang Qutan Recipe (HQQR),a traditional Chinese herbal remedy,has been used for treating hypertension over several years.Objective:This study assesses HQQR’s efficacy for controlling blood pressure among patients with hypertension related to blood stasis,yang hyperactivity and phlegm.Design,setting,participants and interventions:A randomized controlled trial was conducted at the Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,China,from July 2020 to June 2022.Major components of HQQR were identified using thin-layer chromatography and high-performance liquid chromatography.Participants aged18–80 years,exhibiting traditional Chinese medicine syndromes of blood stasis,yang hyperactivity or phlegm,along with grades 1 or 2 hypertension,were randomly categorized into two groups.The intervention group was given HQQR granules alongside conventional hypertension treatment,while the control group was given placebo granules in addition to conventional treatment for 12 weeks.Main outcome measures:The primary outcome was clinic blood pressure,whereas secondary outcomes included metabolic indices (e.g.,homeostasis model assessment of insulin resistance[HOMA-IR],total cholesterol[TC],low-density lipoprotein cholesterol and triglyceride),target organ damage indices (left ventricular mass index and urinary albumin creatinine ratio[UACR]) and inflammation indices(interleukin-6[IL-6]and high-sensitivity C-reactive protein[hs-CRP]).Results:HQQR’s primary components were identified as salvianolic acid B,emodin and ferulic acid.Of the216 participants (108 in each group),compared to the control,the intervention group exhibited significant improvements (P<0.001) in clinic systolic blood pressure ([136.24±7.63]vs[130.06±8.50]mmHg),clinic diastolic blood pressure ([84.34±8.72]vs[80.46±6.05]mmHg),home systolic blood pressure([131.64±8.74]vs[122.36±8.45]mmHg) and home diastolic blood pressure ([78.47±9.53]vs[71.79±6.82]mm Hg).HQQR demonstrated a reduction in ambulatory blood pressure (24-hour systolic blood pressure:[133.75±10.49]vs[132.46±8.84]mmHg and 24-hour diastolic blood pressure:[84.12±8.01]vs[82.11±7.45]mmHg) and an improvement in HOMA-IR ([4.09±1.72]vs[3.98±1.44]),TC ([4.66±1.47]vs[3.75±1.81]mmol/L) and UACR (75.94[5.12,401.12]vs 45.61[4.26,234.26]).Moreover,HQQR demonstrated a decrease in hs-CRP (1.46[0.10,10.53]vs 0.57[0.12,3.99]mg/L) and IL-6 (6.69[2.00,29.74]vs 5.27[2.00,9.73]pg/mL),with no reported side effects (P<0.001).Conclusion:This study highlights the therapeutic potential of HQQR use in ameliorating blood pressure,glycolipid metabolism,and inflammation in patients with hypertension.