Untargeted metabolomics characteristics of nonobese nonalcoholic fatty liver disease induced by high-temperature-processed feed in Sprague-Dawley rats

BACKGROUND Nonalcoholic fatty liver disease(NAFLD)has become one of the most common chronic liver diseases in the world.In our early clinical data and questionnaire analysis of NAFLD,it was found that the body mass index of some patients did not meet the diagnostic criteria for overweight or obesity.The consumption of high-temperature-processed foods such as fried food,hot pot and barbecue is closely related to the occurrence of nonobese NAFLD.Reducing the intake of this kind of food can reduce disease severity and improve prognosis.AIM To explore the untargeted metabolomics characteristics of nonobese nonalcoholic fatty liver disease in Sprague-Dawley rats induced by high-temperatureprocessed feed.METHODS Fifty-four male Sprague-Dawley rats were divided into three groups:The control group received a standard diet;the nonfried soybeans(NDFS)group received 60%NDFS and 40%basic feed and the dry-fried soybeans(DFS)group received 60%DFS and 40%basic feed.Six rats were sacrificed at week 4,8,and 12 in each group.The food intake,body weight,Lee’s index,liver index,serological index and hepatic histopathology were assessed.Untargeted metabolomics characteristics were used to analyze the changes in liver metabolites of rats at week 12.Correlations between metabolites and pathology scores between the DFS and control groups and between the DFS and NDFS groups were analyzed.We selected some of the metabolites,both within the pathway and outside of the pathway,to explain preliminarily the difference in liver pathology in the three groups of rats.RESULTS There were no statistically significant differences in the food intake,body weight,Lee's index or serological index between the DFS group and the control group(P>0.05).At week 8 and week 12,the steatosis scores in the DFS group were significantly higher than those in the other two groups(P<0.05).At week 12,the liver index of the DFS group was the lowest(NDFS group vs DFS group,P<0.05).The fibrosis score in the DFS group was significantly higher than those in the other two groups(P<0.05).The correlation analysis of the liver pathology score and differential metabolites in the DFS and NDFS groups showed that there were 10 strongly correlated substances:Five positively correlated substances and five negatively correlated substances.The positively correlated substances included taurochenodeoxycholate-3-sulfate,acetylcarnitine,20a,22bdihydroxycholesterol,13E-tetranor-16-carboxy-LTE4 and taurocholic acid.The negatively correlated substances included choline,cholesterane-3,7,12,25-tetrol-3-glucuronide,nicotinamide adenine dinucleotide phosphate,lysoPC[16:1(9Z)]and glycerol 3-phosphate.The correlation analysis of the liver pathology score and differential metabolites in the DFS and control groups showed that there were 13 strongly correlated substances:Four positively correlated substances and 9 negatively correlated substances.The positively correlated substances included 4-hydroxy-6-eicosanone,3-phosphoglyceric acid,13-hydroxy-9-methoxy-10-oxo-11-octadecenoic acid and taurochenodeoxycholate-3-sulfate.The negatively correlated substances included lysoPC[16:1(9Z)],S-(9-hydroxy-PGA1)-glutathione,lysoPC[20:5(5Z,8Z,11Z,14Z,17Z)],SM(d18:1/14:0),nicotinamide adenine dinucleotide phosphate,5,10-methylene-THF,folinic acid,N-lactoylglycine and 6-hydroxy-5-methoxyindole glucuronide.CONCLUSION We successfully induced liver damage in rats by using a specially prepared hightemperature-processed feed and explored the untargeted metabolomics characteristics.

Body Roundness Index Is Significantly Associated with Prehypertension and Hypertension in Nonobese Chinese Subjects

Prehypertension and hypertension have been considered significant public health issues worldwide[1].Disproportionate body fat distribution(excess fat stored in the liver and visceral compartment)is a well-established risk factor for prehypertension/hypertension[2].However,the most commonly used anthropometric indicators,such as body mass index(BMI)and waist circumference(WC)。

Nonalcoholic Fatty Liver Disease in Lean/Nonobese and Obese Individuals:A Comprehensive Review on Prevalence,Pathogenesis,Clinical Outcomes,and Treatment

Nonalcoholic fatty liver disease(NAFLD)is the most common liver disease worldwide,with an estimated prevalence of 25% globally.NAFLD is closely associated with metabolic syndrome,which are both becoming increasingly more common with increasing rates of insulin resistance,dyslipidemia,and hypertension.Although NAFLD is strongly associated with obesity,lean or nonobese NAFLD is a relatively new phenotype and occurs in patients without increased waist circumference and with or without visceral fat.Currently,there is limited literature comparing and illustrating the differences between lean/nonobese and obese NAFLD patients with regard to risk factors,pathophysiology,and clinical outcomes.In this review,we aim to define and further delineate different phenotypes of NAFLD and present a comprehensive review on the prevalence,incidence,risk factors,genetic predisposition,and pathophysiology.Furthermore,we discuss and compare the clinical outcomes,such as insulin resistance,dyslipidemia,hypertension,coronary artery disease,mortality,and progression to nonalcoholic steatohepatitis,among lean/nonobese and obese NAFLD patients.Finally,we summarize the most up to date current management of NAFLD,including lifestyle interventions,pharmacologic therapies,and surgical options.

Sialoadenitis progression in nonobese diabetic mice and its correlation with expression of apoptosis-associated proteins in salivary glands and serum IgG levels

Background Sjogren syndrome （SS） is an autoimmune disorder characterized by chronic lymphocytic infiltration and decreased secretion in salivary glands. Apoptosis is one of the possible mechanisms involved in acinar epithelial destruction in SS. The role of apoptosis in the initiation and effect phase of sialoadenitis is still controversial. The aim of this study was to observe the roles of apoptosis-associated proteins and serum IgG levels in sialoadenitis progression in nonobese diabetic （NOD） mice. Methods 2-, 5-, 10-, 15-, 20-week female NOD and matched BALB/c control mice were selected. Saliva and tear flow rate were measured. Serum IgG level was tested by enzyme-linked immunosorbent assay （ELISA）. Number of lymphocyte foci （NLF） in submandibular glands （SMGs） was counted under routine hematoxylin/eosin-stained sections. Expression of Fas, Bcl-2 and procaspase3 proteins as well as apoptotic cells in the SMGs were detected by immunohistochemical staining and by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL） assay respectively. Results Decreased stimulated total flow rate （STFR） and lymphocyte foci in SMGs were first observed in the 10-week NOD group. STFR was negatively correlated with NLF （P〈0.05）. Serum IgG in NOD mice was significantly higher than that of the control group （P〈0.05） and showed a positive correlation with NLF （P〈0.05）. Fas expression in SMGs acinar cells in NOD mice increased with age and was significantly higher compared with that in the control group. Bcl-2 expression and procaspase3 expression in SMG acinar cells in each NOD group were lower compared with those of the age-matched control mice. Conclusion Abnormal expression of Fas and Bcl-2 in the SMGs and higher level of serum IgG may contribute to the initiation of sialoadenitis and cause the glandular destruction in NOD mice.

Effects of Ganoderma lucidum spores on sialoadenitis of nonobese tiabetic mice

Background Sjǒgren syndrome （SS） is a systematic autoimmune disease, on which traditional therapeutic agents show limited effect. More effective agents with longer-lasting and fewer side effects are needed in the clinic. The aim of this study was to investigate the effects of Ganoderma lucindum spores （GLS） on sialoadenitis of nonobese diabetic （NOD） mice. Methods Thirty-two female NOD mice were assigned randomly into 4 groups： low-dose GLS-treated （L-GLS） group and high-dose GLS-treated （H-GLS） group, a dexamethasone group, and a normal saline （NS） control group. Stimulated total saliva flow rate （STFR）, area of lymphocytic infiltration in submandibular glands and ratios of CD4^＋ and CD8^＋ T lymphocytes and B lymphocytes in peripheral blood as well as apoptosis of these subsets and serum IgG level were tested after 10 weeks of treatments. Differences among the groups were analyzed by one-way analysis of variance （ANOVA）, Student-Newman-Keuls Test （SNK） was used between each two groups and a P 〈0.05 was considered statistically significant. Results STFR of the high-dose GLS group increased significantly after a 10-week treatment compared with those of the NS control group （P 〈0.05）. The incidence of sialoadenitis in GLS-treated NOD mice groups showed no significant difference compared with the control group （P 〉0.05）, but the area of lymphocytic foci in both the H-GLS and L-GLS groups decreased significantly to 50% of the NS control group （P 〈0.05）; the ratio of CD4^＋/CD8^＋ T lymphocytes and apoptosis of B lymphocytes of NOD mice with sialoadenitis were less and apoptosis of CD4^＋ and CD8^＋ T lymphocytes were significantly increased compared with the control group （P 〈0.05）. After pretreatment with H-GLS before sialoadenitis onset, the ratio of CD4^＋/CD8^＋ T lymphocyte and the serum IgG levels of NOD mice decreased significantly （P 〈0.05）. Conclusions Pretreatment with H-GLS can relieve symptoms of sialoadenitis in NOD mice. GLS has some protective effects on sialoadenitis in NOD mice through increasing STFR and decreasing the area of lymphocytic foci by regulating the ratio of CD4^＋/CD8^＋ T and apoptosis of B lymphocytes.

Effect of Bushen Qingre Yuyin Decoction on Salivary Secretion, Spleen Index, Submandibular Gland Index, Submandibular Gland Histomorphology, and aqp5 Expression in the Nonobese Diabetic Mouse Model

Objective:The objective of this study was to observe the effect of Bushen Qingre Yuyin Decoction on the spleen index,submandibular gland index,salivary secretion,submandibular gland morphology,and aquaporin 5(AQP5)expression levels in the submandibular gland.Materials and Methods:Fifty 8-week-old nonobese diabetic(NOD)female mice were randomly divided into the following five groups:model;hydroxychloroquine;and high-,medium-,and low-dose Bushen Qingre Yuyin Decoction prescription.The blank group consisted of ten 8-week-old Balb/C rats.Bushen Yuyin Qingre Decoction was administered by gavage at 20,40,and 80 g/kg in the low-,medium-,and high-dose groups,respectively.The hydroxychloroquine group was administered hydroxychloroquine at a dose of 80 mg/kg by gavage.The normal and model groups were administered the same amount of saline.After the different administrations,the amount of salivary secretion in the mice was regularly observed.After 12 weeks,the mice were sacrificed,and their submandibular gland tissues were excised,stained with hematoxylin-eosin,and pathologically scored.AQP5 expression levels in the tissues were detected using real-time polymerase chain reaction.Results:The spleen index of mice in the high-dose group was higher than that in the model group(P<0.05),and the submandibular gland index of the hydroxychloroquine group and all the Bushen Qingre Yuyin Decoction prescription groups was higher than that in the model group(P<0.05).After 6 weeks,the salivary volume of the hydroxychloroquine group and all the Bushen Qingre Yuyin Decoction prescription groups was significantly higher than that of the model group(P<0.05);the amount of lymphocyte infiltration in mice among each administration group was significantly reduced(P<0.05);and the AQP5 expression levels in the submandibular glands of mice had significantly increased in the hydroxychloroquine and high-dose groups(P<0.05),with the high-dose group showing the best effect.Conclusion:Bushen Qingre Yuyin Decoction can effectively increase the spleen and submandibular gland indexes of NOD mice,increase the amount of salivary secretion,reduce lymphocyte infiltration in submandibular gland tissue,and significantly increase AQP5 expression levels,which may be positively correlated with the administered dose.

Fatty liver without a large“belly”:Magnified review of nonalcoholic fatty liver disease in non-obese patients

Nonalcoholic fatty liver disease(NAFLD) is well described as a common cause of chronic liver disease, mostly in the obese population. It refers to a spectrum of chronic liver disease that starts with simple steatosis than progresses to nonalcoholic steatohepatitis and cirrhosis in patients without significant alcohol consumption. NAFLD in the non-obese population has been increasingly reported and studied recently. The pathogenesis of nonobese NAFLD is poorly understood and is related to genetic predisposition, most notably patatin-like phospholipase domain-containing 33 G allele polymorphism that leads to intrahepatic triglyceride accumulation and insulin resistance. Nonobese NAFLD is associated with components of metabolic syndrome and, especially, visceral obesity which seems to be an important etiological factor in this group. Dietary factors and, specifically, a high fructose diet seem to play a role. Cardiovascular events remain the main cause of mortality and morbidity in NAFLD, including in the nonobese population. There is not enough data regarding treatment in non-obese NAFLD patients, but similar to NAFLD in obese subjects, lifestyle changes that include dietary modification, physical activity, and weight loss remain the mainstay of treatment.

Role and function of granin proteins in diabetes mellitus

The granin glycoprotein family consists of nine acidic proteins;chromogranin A(CgA),chromogranin B(CgB),and secretogranin II–VIII.They are produced by a wide range of neuronal,neuroendocrine,and endocrine cells throughout the human body.Their major intracellular function is to sort peptides and proteins into secretory granules,but their cleavage products also take part in the extracellular regulation of diverse biological processes.The contribution of granins to carbohydrate metabolism and diabetes mellitus is a recent research area.CgA is associated with glucose homeostasis and the progression of type 1 diabetes.WE-14,CgA10-19,and CgA43-52 are peptide derivates of CgA,and act as CD4+or CD8+autoantigens in type 1 diabetes,whereas pancreastatin(PST)and catestatin have regulatory effects in carbohydrate metabolism.Furthermore,PST is related to gestational and type 2 diabetes.CgB has a crucial role in physiological insulin secretion.Secretogranins II and III have angiogenic activity in diabetic retinopathy(DR),and are novel targets in recent DR studies.Ongoing studies are beginning to investigate the potential use of granin derivatives as drugs to treat diabetes based on the divergent relationships between granins and different types of diabetes.