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Objective:To examine the effects of apelin-13 on ferroptosis of the C2C12 skeletal muscle cell line induced by a high-iron environment and explore potential underlying mechanisms.Methods:C2C12 cells were cultured in Dulbecco's Modified Eagle Medium(DMEM)and divided into a control group, a ferric citrate(FAC)group, an apelin-13 group, an FAC+ apelin-13 group, a ferroptosis inducer RSL3 group and an FAC+ apelin-13+ RSL3 group.Cell viability was detected by the 3-(4, 5-dimethyl thiazole-2)-2, 5-diphenyl thiazolyl blue(MTT)assay.The intracellular concentrations of total iron and divalent iron were measured by colorimetry; the levels of glutathione(GSH), malondialdehyde(MDA)and intracellular reactive oxygen species(ROS)in cells were detected by an enzyme-linked immunosorbent assay, visible spectrophotometry and a chemifluorescence method, respectively.The ultrastructure of C2C12 cells was examined by transmission electron microscopy.The protein expression of glutathione peroxidase 4(GPX-4), ferritin heavy chain 1(FTH-1), heme oxygenase 1(HO-1)and nuclear factor E2-related factor 2(Nrf-2), were detected by Western blotting.Results:Compared with the FAC group, the FAC+ Apelin-13 group had significantly elevated cell viability(optical density: 0.52±0.06 vs.0.28±0.04, t=7.837, P=0.007)and higher concentrations of GSH(2.41±0.35 vs.0.91±0.12 μmol/g Pro, t=9.778, P=0.003), but significantly decreased levels of ROS(22.06±5.79 vs.52.71±7.28 a. u./mg Pro, t=8.064, P=0.006), MDA(4.63±0.51 vs.9.11±0.84 mmol/mg Pro, t=8.642, P=0.006), total iron(1.53±0.24 vs.3.17±0.55 μmol/g Pro, t=6.135, P=0.013)and divalent iron(0.75±0.08 vs.1.94±0.36 μmol/g Pro, t=5.068, P=0.027), as well as reduced intracellular iron deposition.In the control group and the apelin-13 group, the morphology of the mitochondria was clear and normal.In contrast, the mitochondria in the FAC group had increased membrane density, membrane shrinkage and rupture, vacuolar degeneration, and obvious mitochondrial damage, which were consistent with the morphological characteristics of ferroptosis.Compared with the FAC group, the FAC+ apelin-13 group showed significant improvement in mitochondrial damage.Moreover, compared with the FAC+ apelin-13 group, the cell viability of the FAC+ apelin-13+ RSL3 group was significantly decreased(optical density: 0.23±0.04 vs.0.48±0.06, t=7.642, P=0.007). Compared with the FAC group, the FAC+ apelin-13 group had significantly up-regulated cellular expression of GPX-4(relative expression: 0.96±0.14 vs.0.31±0.07, t=7.712, P=0.008), FTH-1(0.57±0.08 vs.0.27±0.05, t=6.944, P=0.011), and HO-1(0.49±0.07 vs.0.28±0.05, t=6.472, P=0.012), as well as increased nuclear expression of Nrf-2(relative expression: 0.42±0.04 vs.0.19±0.05, t=7.114, P=0.008)with a higher ratio of nuclear expression over total cellular expression[(58.36±5.24)% vs.(36.58±5.32)%, t=5.858, P=0.015]and a higher level of HO-1 protein expression(relative expression: 0.49±0.07 vs.0.28±0.05, t=6.472, P=0.012). Conclusions:Apelin-13 inhibits ferroptosis induced by a high iron environment in C2C12 cells, and the underlying molecular mechanisms may be related to the Nrf-2/HO-1 signaling pathway.
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Objective:To observe the effect of Apelin-13 on diabetes-related sarcopenia in rats model and explore the underlying mechanism.Methods:This experiments were divided into 3 groups.In the normal control group, Wistar rats were fed by ordinary feed, intraperitoneally injected daily with the equal amount of normal saline.In the model+ control group, diabetic-related sarcopenia model was established in rats with fat-fed Goto-Kakizaki rats for 12 weeks and intraperitoneally injected daily with the equal amount of normal saline.In the model+ apelin-13 group, apelin-13(0.1 mol/kg)was intraperitoneally injected into diabetic-related sarcopenia model daily for 12 weeks.The water intake, food intake, and body weight were measured.The fasting blood glucose, fasting insulin and blood lipid levels of rats were measured, and the insulin resistance index was calculated.The wet weight of the rats' gastrocnemius muscle was weighed, and the morphological changes of the gastrocnemius muscle were observed.Western blot was used to detect the protein expressions of phosphatidylinositol-3 kinase(PI3K)and phosphorylated Akt(p-Akt)in the gastrocnemius muscle of rats.Results:Compared with the model+ normal saline group, the model+ apelin-13 group showed that the following parameters were significantly improved.(1)The water intake of rats was significantly decreased at 8, and 12 weeks( F=7.17 and 7.91), and food intake was significantly decreased( F=5.84 and 6.12)and body weight were significantly increased at 8 and 12 weeks( F=5.76 and 6.07)(all P<0.05). (2)The levels of fasting blood glucose were significantly decreased at 8 and 12 weeks( F=8.07 and 8.24, all P<0.05). (3)Serum triglycerides, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, the levels fasting insulin and the insulin resistance index were significantly decreased at week 12( F=5.17, 7.94, 10.27, 8.32, 6.94 and 11.31, all P<0.05); (4)The wet weight of gastrocnemius muscle was significantly increased(0.63±0.04 g and 1.02±0.05 g, t=4.32, P<0.05). (5)Compared with the model+ normal saline group, the model+ apelin-13 group showed the protein expressions of PI3K and p-Akt in the gastrocnemius muscle of rats were significantly up-regulated( t=7.32, 8.07, both P<0.05). Conclusions:Apelin-13 has an inhibitory effect on diabetes-related sarcopenia, and its mechanism may be related to the up-regulation of PI3K and p-Akt expression in skeletal muscle.
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Objective:To explore the possible associated factors and parameters of obstructive sleep apnea hypopnea syndrome (OSAHS) among the elderly in Changsha.Methods:In the investigation of chronic diseases among the elderly in Changsha and the promotion and popularization of community standardized prevention and control, 500 elderly people >65 years old in Yuhua District of Changsha City were selected by multi-stage sampling method. Polysomnography (PSG) was used to detect sleep apnea hypopnea syndrome and collect physical examination indexes. A total of 70 cases met the diagnostic criteria of OSAHS apnea hypopnea index were divided into mild (42 cases), moderate (16 cases) and severe (12 cases). 20 cases of non OSAHS were selected as control group. The general information, PSG level and dynamic blood glucose level were compared between the two groups. Pearson correlation analysis was performed between ambulatory blood glucose level and sleep apnea hypopnea index in OSAHS patients.Results:There were significant differences in body mass index (BMI), girth ratio, smoking history, drinking history, apnea frequency, longest duration of hypoventilation, lowest oxygen saturation, supine apnea index and non supine apnea index between OSAHS group and control group ( P<0.05). The amplitude of blood glucose fluctuation was positively correlated with sleep apnea hypopnea index ( P<0.05). Conclusions:Among the many associated factors on OSAHS, BMI and the ratio of neck circumference to length and body position have the most notable influence. Patients with OSAHS are more prone to blood glucose fluctuations, and gradually worsen with the progression of OSAHS. Therefore, we should take targeted intervention measures.
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Objective@#To observe the effect of dihydromyricetin(DMY) on the M1 polarization of macrophages induced by oxidized low-density lipoprotein(Ox-LDL) and to explore its mechanism.@*Methods@#RAW 264.7 cells were incubated in RPMI1640 medium containing Ox-LDL(50 mg/L) and DMY(40 μmol/L) for 24 h. The effect of JAK/STAT signaling pathway inhibitor AG-490 on DMY was observed.The levels of tumor necrosis factor-α(TNF-α), interferon-γ(IFN-γ), transforming growth factor-β(TGF-β) and interleukin-10(IL-10) in the supernatant of cell culture medium were detected by enzyme-linked immunosorbent assay(ELISA). The protein expressions of inducible nitric oxide synthase(iNOS) and arginase 1(Arg1) were tested by Western blot.@*Results@#Compared with the control group, the Ox-LDL group showed that the protein expression in macrophage of iNOS and levels of TNF-α and IFN-γ in the supernatant of culture medium were significantly increased(t=4.12, 2.89 and 3.44, P=0.023, 0.036 and 0.031), while the protein expression of Arg1 in cells and levels of TGF-β and IL-10 in the supernatant of culture medium were significantly decreased(t=3.71, 2.31 and 2.67, P=0.034, 0.038 and 0.034), and the ratios of p-JAK3/JAK3 and p-STAT5/STAT5 in cells were significantly increased(t=3.93 and 2.65 and P=0.007 and 0.021). Compared with the Ox-LDL group, the Ox-LDL+ DMY group showed that the protein expression of iNOS in cells and levels of TNF-α and IFN-γ in the supernatant of culture medium were significantly decreased(t=5.74, 2.37 and 3.07, P=0.022, 0.031 and 0.033), while the protein expression of Arg1 in cells and levels of TGF-β and IL-10 in the supernatant of culture medium were significantly increased(t=3.94, 2.19 and 2.44, P=0.025, 0.047 and 0.035), and the ratios of p-JAK3/JAK3 and p-STAT5/STAT5 in cells were significantly decreased(t=3.41 and 2.32, P=0.015 and 0.023). Compared with the Ox-LDL+ DMY group, the Ox-LDL+ DMY+ AG-490 group showed that the protein expression of iNOS and levels of TNF-α and IFN-γ in supernatant of culture medium were significantly increased(t=3.14, 3.41 and 3.74, P=0.017, 0.028 and 0.023), while the protein expression of Arg1 and levels of TGF-β and IL-10 were significantly decreased(t=2.87, 2.85 and 3.47, P=0.025, 0.035 and 0.028).@*Conclusions@#Dihydromyricetin inhibits the M1 polarization of macrophages induced by Ox-LDL, and the mechanism may be related with the activation of JAK3/STAT5 signaling pathway.
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Objective To observe the effect of dihydromyricetin(DMY) on the M1 polarization of macrophages induced by oxidized low-density lipoprotein(Ox-LDL) and to explore its mechanism.Methods RAW 264.7 cells were incubated in RPMI1640 medium containing Ox-LDL(50 mg/L) and DMY(40 μmol/L) for 24 h.The effect of JAK/STAT signaling pathway inhibitor AG-490 on DMY was observed.The levels of tumor necrosis factor-α(TNF-α),interferon-y(IFN-γ),transforming growth factor-β(TGF-β) and interleukin-10(IL-10) in the supernatant of cell culture medium were detected by enzyme-linked immunosorbent assay(ELISA).The protein expressions of inducible nitric oxide synthase(iNOS) and arginase 1(Arg1) were tested by Western blot.Results Compared with the control group,the Ox-LDL group showed that the protein expression in macrophage of iNOS and levels of TNF-α and IFN-γ in the supernatant of culture medium were significantly increased(t =4.12,2.89 and 3.44,P=0.023,0.036 and 0.031),while the protein expression of Arg1 in cells and levels of TGF-β and IL-10 in the supernatant of culture medium were significantly decreased(t =3.71,2.31 and 2.67,P =0.034,0.038 and 0.034),and the ratios of p-JAK3/JAK3 and p-STAT5/STAT5 in cells were significantly increased(t =3.93 and 2.65 and P =0.007 and 0.021).Compared with the OxLDL group,the Ox-LDL+DMY group showed that the protein expression of iNOS in cells and levels of TNF-α and IFN-γ in the supernatant of culture medium were significantly decreased(t =5.74,2.37and 3.07,P =0.022,0.031 and 0.033),while the protein expression of Arg1 in cells and levels of TGF-β and IL-10 in the supernatant of culture medium were significantly increased(t =3.94,2.19 and 2.44,P=0.025,0.047 and 0.035),and the ratios of p-JAK3/JAK3 and p-STAT5/STAT5 in cells were significantly decreased(t =3.41 and 2.32,P =0.015 and 0.023).Compared with the Ox-LDL+ DMY group,the Ox-LDL+-DMY + AG-490 group showed that the protein expression of iNOS and levels of TNF-α and IFN-γ in supernatant of culture medium were significantly increased (t =3.14,3.41 and 3.74,P =0.017,0.028 and 0.023),while the protein expression of Arg1 and levels of TGF-β and IL-10 were significantly decreased (t =2.87,2.85 and 3.47,P =0.025,0.035 and 0.028).Conclusions Dihydromyricetin inhibits the M1 polarization of macrophages induced by Ox-LDL,and the mechanism may be related with the activation of JAK3/STAT5 signaling pathway.
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OBJECTIVE@#To study the prevalence and influencing factors of dyslipidemia in the elderly in Changsha.@*METHODS@#Multi-stage randomized cluster sampling method was used to select 3 500 persons aged 65 and over in Changsha. Levels of serum lipids were detected and questionnaire was used to investigate the related factors (such as smoking, drinking, history of chronic diseases).@*RESULTS@#The prevalence rate of dyslipidemia was 43.72%. The abnormal rate of serum triglyceride, cholesterol and low-density lipoprotein was 26.54%, 25.31% and 16.65%, respectively. Multiple logistic regression analysis showed that smoking, drinking, overweight or obesity were risk factors of dyslipidemia.@*CONCLUSION@#The incidence of dyslipidemia is high and the influencing factors are common among the elderly. Community health education will help improve the effect of prevention and control.
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Idoso , Humanos , Consumo de Bebidas Alcoólicas , Colesterol , Sangue , Dislipidemias , Epidemiologia , Lipoproteínas LDL , Sangue , Obesidade , Sobrepeso , Prevalência , Fatores de Risco , Fumar , Inquéritos e QuestionáriosRESUMO
Objective To study the epidemiological tendency and influencing factors for major chronic diseases in the elderly in Changsha,and to provide the scientific basis for the prevention and control strategies for the aged people.Methods Multi-stage randomized cluster sampling method was used in selecting 3135 persons aged 65 and over in Changsha.Using unified questionnaire and face to face asking method to investigate the prevalence of chronic diseases and its influencing factors.Results 95.7％ of questionnaires were valid (3000/3135).The total prevalence rate of chronic disease was 86.33％ in patients aged from 65-96 (74.83±6.63) years,with 85.49％ (1332) in males,and 87.23％ (1258) infemales(x2=1.93,P=0.164).Most of them had 1-3 kinds of chronic diseases,in 1 patient with 13 kinds of chronic diseases at the most.The most common chronic diseases among the elderly were hypertension,heart diseases,osteoarthritis,diabetes mellitus,benign prostatic hyperplasia,and fatty liver.Multivariate logistic regression suggested that age,marital status,smoking,quality of sleep were common influencing factors for hypertension,heart diseases and diabetes mellitus.Conclusions The prevalence of chronic diseases is high and the influencing factors are ubiquitous among the elderly in Changsha community.Community health education should be carried out to improve the effect of prevention and control of chronic diseases.
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Objective To investigate the effect of 17-β-estradiol (17-β-E2) on the expression of glucose transporter 4 (GLUT4) in rat primary culture skeletal muscle cells with insulin resistance (IR) induced by palmitinic acid (PA).Methods Rat skeletal muscle was primarily cultivated.The cells were identified by the technology of immunofluorescence.Cells were inclubated with 0.6 mmol/L palmitinic acid for 24 h to induce IR.The concentration of glucose in the medium was measured.The mRNA and protein expressions of protein kinase B (PKB/Akt) and GLUT4 were measured by real time polymerase chain reaction (PCR) and Western blot respectively.Results Compared the control with PA treated rats,the concentration of glucose in the medium was increased [(3.86±0.64)mmol/L vs.(5.43±0.55) mmol/L,q＝4.13,P＜0.05],uptake of glucose stimulated by insulin was decreased (P＜0.01),the expressions of GLUT4 and phosphorylation PKB/Akt were reduced in the PA group (all P＜0.05).In the 17-β-E2 (100 nmol/L) versus control,the concentration of glucose in the medium was higher [(3.86±0.64) vs.(3.77±0.35)mmol/L,q＝ 4.76,P＜0.05],uptake of glucose stimulated by insulin was increased (P＜0.01) and the expressions of GLUT4 and p-Akt were also increased (all P＜0.05).17-β-E2 (10 nmol/L and 100 nmol/L) reversed the decrease of basal and insulin-stimulated uptake of glucose induced by PA,reversed the decrease of GLUT4 and p-Akt expression induced by PA compared with the PA group (all P＜ 0.05).Conclusions 17-β-E2 inhibits insulin resistance induced by PA in rat primary culture skeletal muscle cells,the mechanism may be correlated with the up-regulation of expression of phospho-Akt and GLUT 4 induced by 17-β-E2.