Effects of ghrelin on mice of obesity and insulin resistance / 医学研究生学报

ObjectiveThe effect of ghrelin on glucose metabolism is still controversial. This study aims to investigate the effects of long-term application of acyl ghrelin (AG) and des-acyl ghrelin (DAG) on insulin resistance and serum inflammatory factor levels by establishing a mouse model of obesity, induced by a high-fat diet.MethodsThirty two male C57BL/6 mice were randomly divided into 4 groups, 8 in each group. Except for the control group, the high fat diet group (HFD), HFD+AG group and HFD+DAG group were given a high-fat diet to induce obesity in mice. Control group: standard feed and an intraperitoneal injection of 10mL isotonic saline were given every day. HFD: high-fat feed and an intraperitoneal injection of 10mL isotonic saline were given every day. HFD+AG group: high-fat diet was fed with 0.8mg AG; HFD+DAG group: high-fat diet was fed with 0.8mg DAG. Intraperitoneal glucose tolerance test (IPGTT) was performed 16 weeks later. The blood glucose was collected from the tail veins at 0min, 30min, 60min and 120min after injection, respectively, the fluctuation curve was drawn, the area under the curve was calculated, and then the epididymal fat index was weighted. Fasting insulin, interleukin 6 (IL6) and TNFα levels were measured by enzyme-linked immunosorbent assay (ELISA). Then the insulin resistance index (HOMA IR) was compared.ResultsAfter 6 weeks of feeding, the weight of the mice in HFD was significantly higher than that of the control group (P<0.05). After 14 and 12 weeks of administration, the mice in the HFD+AG group and the HFD+DAG group had a significant weight loss (P<0.05). The fat mass of the epididymis in the HFD+DAG group [(0.92±0.32)g] was significantly lower than that of the HFD group [(1.08±0.11)g] (P<0.05); the fasting insulin level was significantly lower, too (P<0.05). The insulin resistance index (4.94±1.27, 4.08±1.35), IL6 [(34.82±6.23), (36.90±5.27)pg/mL] and TNFα levels [(73.01±7.75), (69.39±8.43)pg/mL] in the HFD+AG group and HFD+DAG group were significantly lower than those in the HFD group [(81.70±7.53), (45.85±6.41) pg/mL, (81.70±7.53)pg/mL], with statistically significant differences (P<0.05). The serum levels of IL 6 and TNFα in the HFD group were significantly lower than those in the control group (P<0.05).ConclusionLong-term application of AG and DAG could improve the insulin resistance and reduce the inflammation level of the mice induced by a high-fat diet. DAG can also decrease the visceral fat in mice.

Research on the mechanism of gut microbiota in reducing blood glucose of db/db diabetic mice by broad-spectrum antibiotics / 医学研究生学报

Objective The alterations of gut microbiota is closely related to metabolic diseases. The aim of this study was to analyze the effects of antibiotics on glucose metabolism and gut microbiota in mice, and to further explore the mechanism of gut microbiota in reducing blood glucose in db/db diabetic mice by broad-spectrum antibiotics. Methods 16 C57BL/KsJ-db/db mice were randomly divided into antibiotic group and control group with 8 mice in each group. Antibiotic group: broad-spectrum antibiotics(vancomycin 10mg/(kg·d), carbenicillin 50mg/(kg·d), metronidazole 50mg/(kg·d), neomycin 30mg/(kg·d)); Control group: 1% cellulose sodium solution as placebo treatment. Fasting blood glucose and body weights were recorded once a week during the study. At the same time, feces were collected for 16S rDNA gene sequencing analysis. The changes of fasting blood glucose, body weight, the relative abundance of microbiota, Shannon index, Simpson index and GLP-1 were compared between the two groups. Results After 5 weeks of treatment with broad-spectrum antibiotics (Vancomycin , Carbenicillin , Metronidazole , and Neomycin ), fasting blood glucose levels in db/db diabetic mice were significantly decreased (9.59±4.49mmol/L vs 19.71±8.74mmol/L,P=0.016). At the same time, antibiotics can also affect the gut microbiota of mice. The relative abundance of Proteobacteria in mice treated with antibiotics was significantly higher than that in control group (0.471±0.12 vs 0.177±0.12, P<0.05), and the OTUs of Proteobacteria, Enterobacteriaceae, Gamma-proteobacteria, and Enterobacteriales increased in mice treated with antibiotics compared with controls. In addition, we also showed antibiotics could change the diversity of gut microbiota, and the diversity of gut microbiota in antibiotic treated mice decreased significantly (Shannon index 3.135 vs 5.359, P<0.01); Simpson index 0.794 vs 0.946, P<0.01). Conclusion Broad-spectrum antibiotics can significantly reduce the fasting blood glucose level and the diversity of gut microbiota of db / db diabetic mice, and the alterations of gut microbiota may play an essential role in the process of reducing blood glucose by broad-spectrum antibiotics.

Recent studies on the correlation of vitamin B12 and diabetes / 医学研究生学报

Vitamin B12 (VitB12) is one of the essential vitamins in humans and is involved in DNA synthesis and cellular metabolism. Many studies have shown that the lack of VitB12 is closely related to the occurrence and development of diabetes and its complications. Therefore, regular testing and reasonable supplementation of VitB12 can help prevent diabetes complications. The article reviews the relationship between VitB12 and diabetes as well as the application of VitB12 in diabetic patients.

Association analysis of Acyl ghrelin,Des-acyl ghrelin and its ratio with insulin resistance in patients with type 2 diabetes mellitus / 中国综合临床

Objective To explore the association between the different forms of in vivo ghrelin—Acyl ghrelin( AG) ,Des-acyl ghrelin( DAG) and AG/DAG with insulin resistance( IR) in patients with type 2 diabetes mellitus. Methods From June 2017 to November 2017,eighty-three patients with type 2 diabetes hospitalized in (group T2DM) and 40 healthy subjects (group NC) were hospitalized in Jinling Clinical Medicine were selected. Height body mass,blood pressure,blood lipid,glycated hemoglobin ( HbA1c) and fasting blood glucose (FPG),fasting insulin (FINS),and fasting C peptide (F-C-p) were measured,and all subjects were left with fasting serum,and the concentration of AG and DAG were measured by enzyme linked immunosorbent assay (ELISA). The body mass index (BMI),total gastric starvation (T-ghrelin) level,AG/DAG,insulin resistance index ( HOMA-IR) , insulin sensitivity index ( HOMA-IS ) and islet beta cell function ( HOMA- beta ) were calculated. The differences of the above indexes between the two groups were compared, and the relationship between AG,DAG,T-ghrelin,AG/DAG and FPG,HOMA-IR,HOMA-IS and HOMA- beta in T2DM patients were analyzed. Results ( 1) There were no significant difference in SBP、DBP、TC、LDL-C、AG between group NC and group T2DM(P>0. 05). Compared with NC group,the age、TG、BMI、HbA1c、FPG、FINS、HOMA-IR、HOMA-β、AG/DAG were significantly higher in T2DM group ( t=2. 690,-1. 990, 0. 873, 14. 257, 10. 528, Z=2. 885,-3. 483,-2. 284;P<0. 01,P<0. 05) . The HDL-C,F-C-p,HOMA-IS,HOMA-beta,DAG and T-ghrelin in group T2DM were lower than those of NC group( t or Z=0. 477,-3. 812,-3. 395,-4. 4,-2. 916,-2. 834;P<0. 05 or P<0. 01) . ( 2) The correlation analysis showed that there was a positive correlation between AG and FPG in T2DM group (r=0. 252,P<0. 05),DAG and T-ghrelin were negatively correlated with HOMA-IR (r=-0. 394,-0. 384,P<0. 05),and AG/DAG was positively correlated with HOMA-IR (r=0. 394,0. 384,P<0. 05),but is negatively correlated with HOMA-IS (r=-0. 292,P<0. 05). (3) multivariate linear regression analysis showed that FPG in T2DM patients were the influencing factors of AG ( t=2. 865,P<0. 05) ,while FINS and BMI were the influencing factors of DAG( t=-2. 808、-0. 330,P<0. 05) andT-ghrelin( t=-2. 725、-0. 330, P<0. 05) . HOMA-IR and BMI are the influencing factors of AG/DAG ( t=3. 718,3. 069,P<0. 05) . Conclusion The levels of DAG and T-ghrelin in group T2DM were significantly lower than those in the normal population, and was negatively correlated with the insulin resistance index,and the ratio of AG/DAG was closely related to insulin resistance,and the level of AG was mainly affected by fasting blood glucose.

Changes of main components of compound Mailuqishen and its anti-tumor effect in H22 tumor-bearing mice / 吉林大学学报(医学版)

Objective:To determine the changes of the main components after compatibility of compound Mailuqishen(MLQS), and to explore its anti-tumor effect in the H22 tumor-bearing mice.Methods:The contents of ginsenoside and amino acid in MLQS were detected by HPLC, and the content of polysaccharide was detected by phenol-sulfuric acid.A total of 144 female Kunming mice were randomly divided into normal control group, model group, positive control group, low dose of MLQS group, medium dose of MLQS group, high dose of MLQS group, ginseng geng group, ginseng group and antler plate group (n=16).Except normal control group, the mice in the other eight groups were used to establish the H22 tumor-bearing mouse models, then the mice were treated with drugs at next day.The tumor weights, inhibitory rates of tumor, spleen and thymus indexes of the H22 tumor-bearing mice were detected 10 d after administration.The morphological changes of tumor and spleen tissue were examined by HE staining, and the apoptotic rates of H22 tumor cells were tested by flow cytometry.Results:As calculation with the ginseng and antler plate single herb, the contents of ginsenosides, polysaccharides and amino acids in MLQS were significantly higher than those of single herbs (P<0.05).Compared with model group, the inhibitory rates of tumor in various administration groups were significantly increased(P<0.01);the spleen indexes and thymus indexes of the mice in different doses of MLQS groups were significantly increased(P<0.01);the apoptotic rates of tumor cells were markedly increased(P<0.05).Compared with model group, the tumor tissue of the mice in various administration groups was destroyed, the cells were sparse and irregular, and the tumor presented necrotic lesions;the morphology of spleen tissue was normal with discernible fringe, and the lymphocytes arranged densely.Conclusion:The contents of ginsenosides, polysaccharides, and amino acids in compound MLQS are significantly increased compared with those of single herbs, and its anti-tumor effect is stronger than the single herbs.

Study on early screening of high saponin ginseng lines / 中国中药杂志

The study aims at screening the specific bands by PCR, quickly and accurately evaluating the quality of ginseng seeding, accelerating the process of ginseng breeding. Based on the correlation of genetic differences and saponin content between individuals, a pair of specific primer GC1 was screened by PCR. According to the experiment by L16 (45) orthogonal test, a PCR system most suitable for GC1 was established, which came out total 25 μL reaction system containing DNA 2.60 mg•L⁻¹, Mg²⁺ 1.44 mmol•L⁻¹, dNTP 0.19 mmol•L⁻¹, primer 0.32 μmol•L⁻¹ and Taq enzyme concentration 0.076 U•μL⁻¹. By comparing the saponin content and the GC1 PCR electrophoretogram of samples, the ginseng, with 1 200 bp specific band by PCR of GC1, the contents of 9 monosodium saponins and their additions were higher than others, which provided a reliable method for accelerating the process of ginseng breeding. The sequence was sequenced and 99% homologous to glycerol-3-phosphate dehydrogenase.

Brachial-ankle pulse wave velocity correlates to urinaryprotein in patients with type 2 diabetes mellitus / 医学研究生学报

Objective Urinary protein is closely related to the development and progression of major vascular lesionsin diabetic patients.This study was to investigate the correlation between brachial-ankle pulse wave velocity(baPWV) and urinary protein in patients with type 2 diabetes mellitus ( T2DM) . Methods According to the level of 24-hour urinary albumin ( UAL) , 142 T2DM patients were divided into three groups:control (UA<30 mg, n=84), microalbuminuria(UAL<300 mg, n=26), and clinical albuminuria group ( UAL≥300 mg, n=33) .All the patients underwent examination offasting blood glucose, blood lipid, renal function, HbA1c, islet func-tion, C-reactive protein, blood pressure, and BMI.Their baPWV was determined with an Omron atherosclerosis detector and the correla-tion of baPWV with other factors evaluated by Pearson analysis and multivariate linear regression analysis. Results The baPWVwas significantly higher in themicroalbuminuria([1649.19±229.36]cm/s) and clinical albuminuria ([1759.21±291.05]cm/s) than in the control group ([1522.33±248.37]cm/s) (P<0.05).Pearson analysis showed a positive correlation between 24-hour UAL and baPWV ( r=0.347, P<0.01) and multivariate linear regression analysis manifested the same correlation ( P<0.05) . Conclusion The level of urinary protein is closely correlated with baPWV in T2DM patients.