Comparison of the Efficacy of Rosuvastatin Monotherapy 20 mg with Rosuvastatin 5 mg and Ezetimibe 10 mg Combination Therapy on Lipid Parameters in Patients with Type 2 Diabetes Mellitus

BACKGROUND: The apolipoprotein B/A1 (apoB/A1) ratio is a stronger predictor of future cardiovascular disease than is the level of conventional lipids. Statin and ezetimibe combination therapy have shown additional cardioprotective effects over statin monotherapy. METHODS: This was a single-center, randomized, open-label, active-controlled study in Korea. A total of 36 patients with type 2 diabetes mellitus were randomized to either rosuvastatin monotherapy (20 mg/day, n=20) or rosuvastatin/ezetimibe (5 mg/10 mg/day, n=16) combination therapy for 6 weeks. RESULTS: After the 6-week treatment, low density lipoprotein cholesterol (LDL-C) and apoB reduction were comparable between the two groups (−94.3±15.4 and −62.0±20.9 mg/dL in the rosuvastatin group, −89.9±22.7 and −66.8±21.6 mg/dL in the rosuvastatin/ezetimibe group, P=0.54 and P=0.86, respectively). In addition, change in apoB/A1 ratio (−0.44±0.16 in the rosuvastatin group and −0.47±0.25 in the rosuvastatin/ezetimibe group, P=0.58) did not differ between the two groups. On the other hand, triglyceride and free fatty acid (FFA) reductions were greater in the rosuvastatin/ezetimibe group than in the rosuvastatin group (−10.5 mg/dL [interquartile range (IQR), −37.5 to 29.5] and 0.0 µEq/L [IQR, −136.8 to 146.0] in the rosuvastatin group, −49.5 mg/dL [IQR, −108.5 to −27.5] and −170.5 µEq/L [IQR, −353.0 to 0.8] in the rosuvastatin/ezetimibe group, P=0.010 and P=0.049, respectively). Both treatments were generally well tolerated, and there were no differences in muscle or liver enzyme elevation. CONCLUSION: A 6-week combination therapy of low-dose rosuvastatin and ezetimibe showed LDL-C, apoB, and apoB/A1 ratio reduction comparable to that of high-dose rosuvastatin monotherapy in patients with type 2 diabetes mellitus. Triglyceride and FFA reductions were greater with the combination therapy than with rosuvastatin monotherapy.

Effect of ghrelin O-acyltransferase inhibition by small interfering RNA on hepatocyte fatty degeneration and related mechanism of action / 临床肝胆病杂志

Objective To investigate the effect of inhibition of ghrelin O-acyltransferase (GOAT) by small interfering RNA (siRNA) on hepatocyte fatty degeneration and related mechanism of action.Methods Human LO2 hepatocytes were treated with free fatty acid (FFA)to induce hepatocyte fatty degeneration.LO2 hepatocytes were treated with FFA and siRNA-GOAT alone or in combination and then divided into normal control (NC) group (treated with phosphate buffered saline alone),siRNA-GOAT group (treated with siRNA-GOAT at a final concentration of 10 nm),FFA group (treated with FFA at a final concentration of 1 mm),and FFA + siRNA-GOAT group (treated with FFA at a final concentration of 1 mm and siRNA-GOAT at a final concentration of 10 nm).Oil red O staining was performed for hepatocytes to identify lipid droplets;the triglyceride (TG) test kit was used to measure the lipid level in LO2 hepatocytes;Western blot,qRT-PCR,immunofluorescent staining,and electron microscopy were used to measure autophagy;ELISA and RT-PCR were used to measure the levels of tumor necrosis factor-α (TNFα) and interleukin-6 (IL-6);ELISA was used to measure the changes in the levels of mammalian target of rapamycin (mTOR),phosphorylated mTOR (p-mTOR),AMP-activated protein kinase (AMPK),and phosphorylated AMPK.A one-way analysis of variance was used for comparison between multiple groups,and the least significant difference t-test was used for further comparison between any two groups.Results Compared with the FFA group,the FFA + siRNA-GOAT group had a significant reduction in the formation of lipid droplets and a significantly lower TG level (P ＜0.001).Compared with the FFA group,the FFA + siRNA-GOAT group had significant reductions in the protein and mRNA expression of TNFα and IL-6 (all P ＜ 0.005).The siRNA + GOAT group had significantly higher mRNA expression of LC3-Ⅱ and Beclin-1 than the NC group (all P ＜0.001).The FFA + siRNA-GOAT group had significantly higher mRNA expression of LC3-Ⅱ and Beclin-1 than the FFA group (all P ＜0.001).The siRNA + GOAT group had significantly higher protein expression of LC3-Ⅱ and Beclin-1 than the NC group (all P ＜ 0.05).The FFA + siRNA-GOAT group had significantly higher protein expression of LC3-Ⅱ and Beclin-1 than the FFA group (all P ＜ 0.05).Immunofluorescent staining showed that compared with the FFA group and the siRNA-GOAT group,the FFA + siRNA-GOAT group had a significant increase in the expression of endogenous LC3-Ⅱ in LO2 hepatocytes.Electron microscopy showed that compared with the FFA group,the FFA + siRNA-GOAT group had a significant increase in the expression of autophagosome.After the LO2 hepatocytes were treated by autophagy inhibitors siRNA-ATG5 and 3-MA or an autophagy stimulant,rapamycin,there was a significant difference in TG level between the FFA + siRNA-ATG5 group and the FFA + siRNA-GOAT group (P ＜ 0.001),as well as between the FFA + 3-MA group and the FFA + rapamycin group (P ＜ 0.001).The FFA + siRNA-GOAT group had a significantly higher level of LC3-Ⅰ/Ⅱ than the FFA + siRNA-ATG5 group (P ＜0.05),and the FFA + rapamycin group had a significantly higher level of LC3-Ⅰ/Ⅱ than the FFA + 3-MA group (P ＜ 0.05).Compared with the FFA group,the FFA + siRNA-GOAT group had significantly higher protein expression of p-AMPK (P ＜ 0.05) and significantly lower protein expression of p-rmTOR (P ＜ 0.05).Conclusion GOAT inhibition by siRNA can upregnlate autophagy and alleviate hepatocyte fatty degeneration,possibly by regulating the AMPK/mTOR pathway.

In Vitro Effect of Fatty Acids Identified in the Plasma of Obese Adolescents on the Function of Pancreatic β-Cells

BACKGROUND: The increase in circulating free fatty acid (FFA) levels is a major factor that induces malfunction in pancreatic β-cells. We evaluated the effect of FFAs reconstituted according to the profile of circulating fatty acids found in obese adolescents on the viability and function of the murine insulinoma cell line (mouse insulinoma [MIN6]). METHODS: From fatty acids obtained commercially, plasma-FFA profiles of three different youth populations were reconstituted: obese with metabolic syndrome; obese without metabolic syndrome; and normal weight without metabolic syndrome. MIN6 cells were treated for 24 or 48 hours with the three FFA profiles, and glucose-stimulated insulin secretion, cell viability, mitochondrial function and antioxidant activity were evaluated. RESULTS: The high FFA content and high polyunsaturated ω6/ω3 ratio, present in plasma of obese adolescents with metabolic syndrome had a toxic effect on MIN6 cell viability and function, increasing oxidative stress and decreasing glucose-dependent insulin secretion. CONCLUSION: These results could help to guide nutritional management of obese young individuals, encouraging the increase of ω-3-rich food consumption in order to reduce the likelihood of deterioration of β-cells and the possible development of type 2 diabetes mellitus.

The influence of cholecystokinin-octopeptide on oxidative stess in islet beta cells injured by free fat acids / 重庆医学

Objective To observe the effect of cholecystokinin-octopeptide(CCK-8) on oxidative stress and cell proliferation in mice islet β cells (NIT-1 cells) injured by high concentration free fat acids .Methods In vitro cultured NIT-1 cells were divided into 3 groups ,they were control group ,FFAs group (add 0 .25 mmol/mL of oleinic acid + 0 .25 mmol/mL of palmic acid) and CCK-8 group (add FFAs and 1 × 10 - 8 mmol/L of CCK-8 simultaneously) .Cell morphologies were observed ;NIT-1 cells proliferations were detected by M TT method ,and apoptosis rates were measured by flow cytometry ;The levels of T-AOC ,GSH-Px ,CAT ,SOD and MDA in supernatant were also measured .Results There were less cell debris in CCK-8 group than FFAs group(all P< 0 .01) ;the OD570 value of CCK-8 group was significant higher than FFAs group(P< 0 .01) ,and the 72 h CCK-8 group was higher than 48 h CCK-8 group(P< 0 .01) .Compared with FFAs group ,the levels of CAT ,T-AOC ,SOD and GSH-Px in CCK-8 group were in-creased and the concentration of MDA was decreased obviously(P< 0 .05) ,the levels of CAT ,SOD in 72 h CCK-8 group were high-er than 48 h CCK-8 group ,MDA was lower than 48 h CCK-8 group(P< 0 .05) .Conclusion CCK-8 could protect islet β cells injury from FFAs through anti-oxidative stress mechanism and promote NIT-1 cells proliferation .

The effect of therapeutic life style on serum FFA and constitution index in patients with coronary heart disease complicated metabolic syndrome / 重庆医学

Objective To investigate the level of serum free fatty acid (FFA )after improving the life style in patients with coronary heart disease complicated metabolic syndrome and the effect of therapeutic life style on traditional risk factors of coronary artery disease.Methods A total of 395 patients with coronary heart disease complicated metabolic syndrome were recruited.Pa-tients were divided into intervention group (group A,conventional drug therapy+ intensive life style intervention,n=97)and non-intervention group (group B,conventional drug therapy,n=38)according to the scores of life style.Serum free fatty acid (FFA) was determined by ELASA.The scores of life style was obtained bylife style questionnaire.Results (1)The serum FFA of pa-tients with coronary heart disease complicated metabolic syndrome were positively related to waist circumference and waist-high-ra-tio.(2)Waist circumference,BMI and FFA of group A were significantly lower than those in group B after therapeutic life style in-tervention(P <0.05).(3)Compared with the baseline,the constitution index and FFA in group A were significantly lower after 6-months therapeutic life style intervention(P <0.05).Conclusion Therapeutic life style can reduce the level of FFA and constitution index of the patients with coronary heart disease complicated metabolic syndrome.

The changes of LCHAD in preeclampsia with different clinical features and the correlation with NADPH P47-phox, p38MAPK- α, COX-2 and serum FFA and TG / 中华妇产科杂志

Objective To investigate the changes of fatty acid oxidase in the placenta of preeclampsia cases with different clinical features, and the relationship with oxidative stress and inflammatory response. To study the correlation of serum free fatty acid (FFA) and triglycerides (TG) level in early second trimester with the molecular changes of the long-chain fatty acid oxidase in the third trimester. Methods This was prospective cohort study, in which cases with singleton pregnancies who archived in Haidian Maternal and Children′s Hospital, Beijing, from January 1st 2012 to May 31st, with regular prenatal care were included. Doppler ultrasound was used for screening for the presence of early diastolic notch of uterine artery at 22-24 weeks of gestation. All the 101 cases with the early diastolic notch of uterine artery were included as the notch group, and 377 cases without the early diastolic notch of uterine artery were included as the non-notch group. The perinatal outcomes and the incidence of hypertensive disorders in pregnancy of the two groups were observed. The serum level of FFA and TG was tested, and the mRNA and protein expression of long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD), P47-phox subunit of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, p38 mitogen-activated protein kinase α (p38MAPK-α) and cyclooxygenase-2 (COX-2) were detected using real-time quantitative PCR and western blot. The relationship between serum level of FFA and TG and the mRNA and protein expression of LCHAD, NADPH P47-phox,p38MAPK-α and COX-2 of the placental tissue specimens were analyzed. Results (1) In the notch group, there were 9 cases of early-onset preeclampsia,15 cases of late-onset preeclampsia and 10 cases of gestational hypertension;and there were 8 cases of late-onset preeclampsia and 18 cases of gestational hypertension in the non-notch group. 15 cases with normal blood pressure in each group were randomly selected as the control group.(2)The serum level of TG of cases of early-onset preeclampsia, late-onset preeclampsia and gestational hypertension in the notch group were(2.0±0.8),(1.8±0.6)and (1.9±0.7)mmol/L, and that of FFA were(0.68±0.26),(0.52±0.10)and(0.52±0.17)mmol/L, respectively. The serum level of TG of cases of late-onset preeclampsia and gestational hypertension in the non-notch group were(1.6±0.6)and(1.4±0.4)mmol/L, and that of FFA were(0.49±0.11)and(0.48±0.05)mmol/L, respectively. The serum level of TG and FFA in the control group were(1.4±0.5)and(0.52±0.06)mmol/L, respectively. The TG level of the notch group was higher than that of the control group, and the difference was statistically significant (P 0.05).(6)The mRNA expression of placental LCHAD in the early-onset preeclampsia in the notch group was significantly negatively correlated with the mRNA expression of placental NADPH P47-phox and COX-2 (r=- 0.877,-0.762, P<0.05). The mRNA expression of placental LCHAD in the control group was significantly negatively correlated with the mRNA expression of placental COX-2 (r=- 0.565, P<0.01). The protein expression of placental LCHAD in the early-onset preeclampsia in the notch group was significantly negatively correlated with the protein expression of NADPH P47-phox (r=- 0.818, P<0.01). The protein expression of placental LCHAD in the control group was significantly negatively correlated with the protein expression of COX-2 (r=- 0.502,P<0.01). Conclusions The placental mRNA and protein expression of long-chain fatty acid oxidation enzymes were different in different clinical features of preeclampsia, which were reduced more obviously in the early-onset preeclampsia in the notch group than that of the late-onset preeclampsia in the notch group, and were negatively correlated with the elevated serum FFA level, significantly enhanced oxidative stress and inflammatory response, but with no correlation with serum TG level.

The levels and clinical value of serum free fatty acid in patients with ankylosin g spondylitis / 中华风湿病学杂志

Objective To study the association between serum free fatty acid (FFA) and ankylosing spondylitis (AS).Methods According to the classification criteria,a total of 90 newly diagnosed AS patients,223 healthy individuals and 82 patients with non-inflammatory diseases were divided into three groups,and biochemistry and immunology biomarks were measured in all individuals.One-Way analysis of variance (ANOVA) test was used to compare the difference between the three groups in the serum indexes,and Logistic regression analysis was used to identify AS risk factors associated with AS.Results There were no significant differences in gender,age,body mass index (BMI),white blood cells (WBC),high-level data link control (HDL-C),low-density lipoprotein control (LDL-C),lipoproteins [Lp (a)],alkaline phosphatase (ALP) and TG in the three groups,and our results showed that serum FFA was statistical different between the three groups (F=24.191,P＜0.01),the serum level of FFA in patients with AS was higher compared with patients with noninflammatory diseases and healthy controls [(0.48 ±0.18) mmol/L,(0.28 ±0.09) mmol/L,(0.29±0.16) mmol/L;t=-5.969,P＜0.01;t=5.106,P＜0.01].Seral IgA,IgG,IgM levels,ESR and CRP were statistically different between the three groups (F=14.870,P＜0.01;F=16.464,P＜0.01;F=4.124,P=0.018;F=97.002,P＜0.01;F=22.069,P＜0.01).Gender,age,BMI,serum IgA,IgM,ALP,HDL-C,LDL-C,Lp(a) and TG levels were not associated with AS by logistic regression analysis.However,serum IgG level,ESR and CRP were associated with AS [OR05％CI):1.659(1.032,2.660),P=0.037;OR05％CI):1.340(1.005,1.787),P=0.046;OR05％CI):1.820 (1.025,3.232),P=0.041],and there is an association between FFA and AS was observed in logistic regression analysis (OR=1.132,95％CI:1.014-1.421,P=0.033).Conclusion We suggest that incre-ased FFA is closely associated with AS,and may be an underlying risk factor for AS.

Establishment and Application of Hyperinsulinemic-Euglycemic Clamp Technique in Conscious Rats / 天津医药

Objective To establish the hyperinsulinemic-euglycemic clamp(HEC) technique in conscious rats, and to explore the effect of acute infusion of lipid on glucose infusion rate (GIR) in rats. Methods Ten SD rats were random-ly divided into two groups, 5 rats for each group. The right jugular vein and left carotid artery were catheterized and under-went a HEC with infusion of lipid (intralipid group) for 6 hours, and with continuing infusion of 5%glucose (control group). The plasma levels of free fatty acid(FFA) and GIR were measured by HEC method. Results The level of FFA concentration increased by 17.6-fold, and GIR was reduced by 27%in the intralipid group compared to those of control group (P＜0.001). Conclusion The rat model of HEC has been successfully established by intravenous intralipid infusion, which can be con-firmed by HEC technique.

The Correlation Analysis of CHGA and ATGL in Patients of Type 2 Diabetes Mellitus with Nonalcoholic Fatty Liver Disease / 天津医药

Objective To investigate the relationship between plasma levels of chromogranin A (CHGA) and adi-pose triglyceride lipase (ATGL) in patients with type 2 diabetes (T2DM) combined non-alcoholic fatty liver disease (NAFLD). Methods The plasma levels of CHGA and ATGL were assayed by enzyme-linked immunosorbent assay (ELISA) in T2DM patients with NAFLD (group A, n=74), T2DM without NAFLD (group B, n=76), and normal group (group NC, n=75). The correlation between CHGA, ATGL and other metabolic index was analyzed. Results The plasma level of CHGA was significantly higher in group A (83.15±9.46) and group B (70.90±2.75) than that of group NC (46.74±8.15, P<0.01), and the level of CHGA was significantly higher in group A than that of group B (P<0.01). The plasma level of ATGL was sig-nificantly lower in group A (21.36±13.42) and group B (40.29±22.83) than that of group NC (72.30±26.41, P<0.01), and the level was lower in group A than that of group B (P<0.01). There was a negative correlation between the plasma CHGA, AT-GL and carbohydrate oxidation rate in group A. There was a positive correlation between fasting insulin (FINS), insulin resis-tance index (HOMA-IR), free fatty acid (FFA) and fat oxidation rate in group A. There was a negative correlation between plasma level ATGL and body mass index (BMI), FINS, cholesterol (TC), triglyceride (TG) and HOMA-IR, meanwhile, it was positively correlated with FFA. The multiple stepwise regression analysis showed that FINS, ATGL and FFA were indepen-dent variables for CHGA. The Logistic regression analysis showed that plasma levels of CHGA, ATGL and FFA were the in-dependent predictors of T2DM with NAFLD. Conclusion The plasma levels of CHGA and ATGL are closely correlated with substance and energy metabolism, and the interaction between them may play an important role in the pathogenesis of T2DM with NAFLD .

Correlation of free fatty acid and blood lipids in early second trimester and uterine artery notch in pre-eclampsia / 中华围产医学杂志

Objective To analyze the heterogeneous variation of serum free fatty acid (FFA) and lipids during early second trimester in women with or without uterine artery notch in pre-eclampsia (PE).Methods This is a prospective cohort study of 4 000 women with singleton pregnancies registered in early pregnancy and in whom regular check-ups were performed in Haidian Maternal & Child Health Hospital.Blood specimens were collected at gestational age 14-18 weeks at the same time of screening for Down's syndrome.One hundred and one cases with early diastolic notch of the uterine artery were included in the N+ group,and 172 cases without notch but at high risk of PE were included in the N-group at 22-24 weeks.In addition,205 women who were selected randomly at a ratio of 1 ∶ 5,without notch or PE high-risk factors,were also included in the N group.Both groups were subgrouped according to the outcomes of pregnancy complications:early-onset PE group EPE,late-onset PE (LPE),gestational hypertension (GH) group,gestational diabetes mellitus (GDM) group with normal blood pressure,and no complications (NC) group.The variation in FFA and other lipid metabolism indicators in the PE subgroups were compared and analyzed by two independent-sample t-test,one-factor analysis of variance,Chi-square test (or Fisher's exact) and Logistic regression.Results History of PE and pre-hypertension at first visit differed significantly between the N+ and N-groups [3.9％ (4/101) vs.0.8％ (3/377),x2=5.52,P＜0.05; pre-hypertension at first visit,42.2％ (43/101) vs.25.7％ (97/377),x2=10.91,P＜0.05].In the N+ group,23.8％ (n=24) of women had PE,of which 37.5％ (n=8) were early onset.In the N group,2.1％ (n=8) had PE,and all were late onset.The incidence of PE differed significantly between the N+ and N-groups (x2=59.72,P＜0.05).In the N+ group,FFA gradually decreased among the ePE,IPE,GH and NC groups [(0.68±0.27),(0.58±0.21),(0.57±0.21) and (0.49±0.19) mmol/L,F=2.78,P＜0.05]; Multivariate regression analysis showed that FFA (OR=135.68,95％CI:3.78-4 873.00) and PE history (OR=123.25,95％CI:9.27-i 638.00) were risk factors of ePE.Pre-hypertension at registration (OR=4.69,95％CI:2.08-10.58) and pre-pregnancy body mass index (BMI) 24-28 (OR=3.69,95％CI:1.26-10.83) were risk factors ofGH.FFA (OR=9.08,95％CI:2.49-33.01) and pre-pregnancy BMI ≥ 28 (OR=5.08,95％CI:2.16-11.92) were risk factors for GDM.Conclusions Serum FFA and TG levels in early second trimester are correlated with PE,especially the early-onset PE.The onset of PE is heterogeneous and affected by many factors,and occurs in patients with or without early diastolic notch of the uterine artery in the second trimester.Patients with notch are more likely to have early-onset PE,which is correlated with blood FFA and TG levels.

Plasma free fatty acids composition and its association with nonalcoholic fatty liver disease / 重庆医学

Objective To study the relationship between plasma free fatty acids composition and the incidence of nonalcoholic fatty liver disease(NAFLD) .Methods By the design of case‐control study ,105 patients with NAFLD as cases and 110 healthy peo‐ple as controls were enrolled into the study .Plasma free fatty acid levels were determined by gas chromatography .Results High level of plasma palmitic acid(C16 :0)(OR=1 .769) was the risk factors of NAFLD ,while plasma levels of linoleic acid(C18 :2 n‐6) (OR=0 .855) and arachidonic acid(C20 :4 n‐6)(OR=0 .181)were negatively associated with the incidence of NAFLD .Conclusion These findings suggest that a proper ratio of diet fatty acids intake may reduce the risk of NAFLD .

Effects of Puerarin on Glucose, Lipid Metabolism and Gastric Motility in Early Period Type 2 Diabetic Rats / 中国中医药信息杂志

Objective To investigate the effects of Puerarin on glucose and lipid metabolism and gastric motility in early period type 2 diabetic (T2DM) rats.Methods Rat model of T2DM was established by high fat-sugar diet fed and low-dose streptozotocin-treated. SD rats were divided randomly into normal control group (NC), normal+Puerarin group (NP), diabetes control group (DC) and diabetes+Puerarin group (DP). NP and DP rats were given Puerarin 400 mg/(kg?d) once per day for 5 weeks, NC and DC rats were given PBS. Half time of gastric emptying and emptying rate were evaluated by SPECT. The serum level of FBG, GSP, FFA, TC, TG and INS were measured by kit.Results Compared with NC group, DC rats had higher FBG, FFA, TC, GSP, TG and emptying rate, but INS and half time of gastric emptying decreased significantly (P<0.05,P<0.01). Compared with DC group, TG, GSP, FFA and emptying rate of DP rats were reduced (P<0.05), but had more half time of gastric emptying (P<0.05). The results of multivariate stepwise regression analysis showed that FBG related to half time of gastric emptying.Conclusion Type 2 diabetic rats have faster gastric motility, higher blood glucose and lipid. Puerarin might improve the disorders of GSP, TG, FFA and gastric emptying in diabetic rats.

Mechanism of insulin resistance of islet beta-cells in rat after long term lipid infusion / 中国医师杂志

Objective To study the changes and mechanism of the function of islet βcells and insulin signal transduction molecules in rats after long-term period lipid infusion.Methods Thirty SpragueDawley (SD) rats were randomly divided into free fatty acid (FFA) and normal saline (NS) groups.Catheters were implanted under pentobarbital anesthesia in the right atrium via the jugular vein and the left carotid artery.A technique for a 72-h infusion in unrestrained rats was used for triglyceride and heparin or saline infusion.The infusion period was started on day 2 after surgery.After 72-h infusion,fasting serum insulin (Ins) and FFA in the blood were determined.The glucose infusion rat (GIR) was measured by hyperinsulinemia euglycemic clamp to evaluate the peripheral insulin resistance.The intravenous glucose tolerance test (ivgtt) and islet cell perifusion was conducted to evaluate the function of islet β-cell.The rats in two groups were sacrificed,and the pancreatic islets were isolated and collected.The levels of malondialdehyde (MDA) and reduced glutathione hormone (GSH) were detected in pancreatic tissues.The expressions of insulin receptor substrate-1 (IRS-1),insulin receptor substrate-2 (IRS-2),and glucose transporter-2 (Glut2)gene in islets were detected by real-time polymerase chain reaction (PCR).Results (1)The serum FFA concentration in the FFA group was higher than in NS group [(1.56 ± 0.21) mmol/L vs (0.65 ± 0.12)mmol/L,P ＜0.01].(2)The GIR was decreased significantly in FFA group compared with NS group(P ＜0.01).(3)The glucose that stimulated insulin secretion was decreased in the FFA group.(4)The levels of MDA were significantly higher in FFA group [(1.62 ± O.18) mmol/mg prot vs (0.76 ± 0.15) mmol/mg prot,P ＜0.01].The levels of GSH were lower in FFA group [(22.54 ±2.66) mg/g prot vs (36.58 ± 3.02) mg/g prot,P ＜ 0.01].(5) The gene cxprcssion of IRS-1 in islets was significantly decreased by [(36.8±1.8)％,P ＜0.01],and the expression of IRS-2 and Glut-2 was decreased by [(29.6±1.2) ％] and [(58.7 ± 2.1) ％] in FFA group,respectively(all P ＜0.01).Conclusions Lipid infusion in long time decreased the secretion of insulin and impaired the expression of insulin signal transduction molecules in islet βcells.

The correlation between free fatty acid and insulin resistance in ischemic stroke patients / 中华检验医学杂志

Objective To explore the level of free fatty acid (FFA)in ischemic stroke (IS) patients and its correlation with insulin resistance (IR).Methods This study was case-control study.Patients who were diagnosed with IS were followed up from June,2011 to September,2012 in Tianjin Third Central Hospital.The 180 IS patients were divided into large infarct(57),middle infarct(63)and small infarct (60) according to the area of infarct.At the same time,60 healthy persons were selected as control group.The plasma FFA,fasting insulin (FINS),fasting blood glucose (FBG),homeostasis model assessment (HOMA)-IR,triacylglycerol (TG),total cholesterol (TC),high density lipoprotein cholesterol-C (HDL),low density lipoprotein cholesterol-C(LDL),lipoprotein A1 (APOA1) and lipoprotein B (APOB) in 180 IS patients were measured and compared with that of control group.The correlation between FFA and IR was analyzed by using pearson linear correlation.The relationship between FFA and HOMA-IR was analyzed by using multiple stepwise multivariate analysis.The t test and analysis of variance were used for statistical analysis.Results Compared to normal control,the level of FFA,FINS,FBG,FIB,TG,TC,LDL,APOB and HOMA-IR in IS patients were higher,the difference was statistically.The level of HDL,APOA1 in IS patients were lower than that of normal control.The level of FFA,FINS,HOMA-IR,FBG,TG,TC,HDL,LDL,APOA1 and APOB in IS patients related to the area of infarct.The level of FFA,FINS,HOMA-IR,FBG,TG,TC,LDL,APOB in large infarct were higher than those in middle infarct and small infarct.The level of HDL,APOA1 in large infarct were lower than those in middle infarct and small infarct,the difference were statistically.In IS patients,FFA had positive correlation to FINS,FBG and HOMA-IR (r =0.870,0.497,0.792,P ＜ 0.001),but had no correlation to age (r =0.008,P ＞ 0.05).In IS patients,HOMA-IR was dependent variable,but FFA,FINS,FBG,TG,TC,HDL,APOA1 and APOB were independent variable.FFA was independent risk factor to HOMA-IR,besides FINS,FBG,TC,LDL and APOB.Conclusion FFA elevates in IS patients,FFA has correlation to IR and it is a dependent risk factor to IS patients.

Relationships between concentrations of free fatty acid in serum and oxidative-damage levels in placental mitochondria and preeclamupsia / 中华妇产科杂志

Objective To investigate the relationships between concentrations of free fatty acid (FFA) in maternal serum and oxidative damage levels in placental mitochondria and preeclampsia ( PE)-Methods A total of 60 women with PE and 60 normal pregnant women as control participated in this study.All were admitted to Fujian Maternity and Child Health Hospital for delivery from August 2010 to May 2011.Patients with PE were divided into early-onset group ( n =30,presented at ＜ 34 weeks of gestation ) and late-onset group ( n =30,presented at ≥ 34 weeks of gestation),with 30 normal pregnant women as early control group ( ＜ 34 weeks of gestation ) and 30 as late control group ( ≥34 weeks of gestation).Improved copper agent colorimetry was used to detect FFA in maternal serum Ultraviolet colorimetry was used to detect glutathione peroxidase (GPX) and catalase (CAT) activity in maternal placenta and malondialdebyde (MDA) and permeability transiton (PT) pore in placental mitochondria.Total superoxide dismutase (SOD) assay kit-WST was used to detect SOD activity in placenta.Real-time fluorescent quantitative PCR was used to detect mitochondrial DNA (mtDNA) expression in placenta.Results ( 1 ) Maternal serum FFA was ( 1.6 ±0.5 ) mmol/L in early-onset PE group and ( 1.5 ± 0.4) mmol/L in lateonset PE group,significantly elevated as compared to ( 1.0 ± 0.5 ) mmol/L in early control group and (0.9 ±0.5) mmol/L in late control group (P ＜ 0.05 ). However,no significant difference was found between early-onset and late-onset PE groups (P ＞ 0.05 ).(2) The mean placental GPX,CAT and SOD activity were significantly decreased in the early-onset PE group [ (47 ±6),( 19 ±5),(62 ± 13) U/mg]and late-onset PE group [ (67 ±6),(20 ±4),(96 ± 17) U/mg] as compared to late control group [ (80 ±3),(55 ± 3 ),( 123 ± 19 ) U/mg],respectively ( P ＜ 0.05 ).(3) The mean placental mitochondria MDA was significantly elevated in the early-onset PE group [ (115 ± 22) nmol/mg] and late-onset PE group [(90±17) nmol/mg] as compared to late control group [(52 ± 11) nmol/mg,P ＜0.05].The mean absorption value that present the permeability of placental mitochondria PT pore was significantly elevated in the early-onset PE group (0.086 ±0.013) and late-onset PE group (0.069 ±0.014) as compared to late control group (0.052 ± 0.0 12,P ＜ 0.05 ).The mean placental mtDNA expression was significantly elevated in the early-onset PE group (3.0 ±0.7) and late-onset PE group (2.8 ±0.7) as compared to late control group ( 2.6 ± 0.6,P ＜ 0.05 ).( 4 ) The mean placental mitochondria MDA concentration correlated positively with the concentrations of FFA in maternal serum in the early-onset PE group ( r =0.703,P ＜0.05 ) and late-onset PE group (r =0.457,P ＜ 0.05 ),and negatively with placental antioxidant enzyme in the early-onset PE group ( r =- 0.652,- 0.787,- 0.952 ; P ＜ 0.05 ) and late-onset PE group ( r =-0.378,-0.689,-0.854; P＜0.05).Conclusions Increased FFA in maternal serum and high levels of oxidative damage in placental mitochondria may be involved in the pathogenesis of preeclampsia.Increased FFA in serum and decreased activity of antioxidant enzyme in placenta may contribute to oxidative damage levels in placental mitochondria in women with PE.

Study on the oxidative stress and inflammation in trophoblast cells stimulated by different chain length fatty acids / 中华妇产科杂志

Objective To investigate the oxidative stress and inflammation in trophoblast cells stimulated by different chain length fatty acids.MethodsSerum-free trophoblast cells cultured in vitro were divided into five groups,which were incubated with DMEM medium without free fatty acid (F-FFA),short chain fatty acids (SC-FFA),medium chain fatty acids (MC-FFA),long chain fatty acids (LC-FFA),very long chain fatty acids (VLC-FFA).Then cells in each group were stimulated by DMEM medium,reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor (apocynin) and p38 mitogen-activated protein kinases (p38MAPK) inhibitor (SB203580) and were subdivided as each FFA plus-DMEM group, plus-NADPH-Ⅰ and plus-p38MAPK-Ⅰ groups.Expressions of mRNA and protein of p38MAPK and cyclooxygenase 2 (COX-2) in trophoblast cells were detected by real-time PCR and western blot.Results (1) The mRNA expression of p38MAPK in LC-FFA + DMEM,VLC-FFA + DMEM,LC-FFA + NADPH-Ⅰ,LC-FFA + p38MAPK-Ⅰ,VLC-FFA + NADPH-Ⅰ,VLC-FFA + p38MAPK-Ⅰ group were 4.56 ±0.28,22.65 ±2.40,0.87 ±0.06,1.02 ±0.15,19.87 ± 1.93,10.22 ±0.75 separately,and the protein expressions were 0.79 ± 0.02,0.93 ± 0.10,0.43 ± 0.06,0.44 ± 0.19,0.79 ± 0.10,0.81 ±0.14.Compared with other groups,the mRNA and protein expressions of p38MAPK in LC-FFA + DMEM,VLC-FFA + DMEM group were increased ( P ＜ 0.05 ).Compared with LC-FFA + DMEM group,mRNA and protein expressions of p38MAPK in LC-FFA + NADPH-Ⅰ and LC-FFA + p38MAPK-Ⅰ group were significantly decreased (P ＜ 0.05 ).Compared with VLC-FFA + DMEM group,mRNA and protein expressions of p38MAPK had no difference in VLC-FFA + NADPH-Ⅰ group (P ＞ 0.05 ),mRNA expression of p38MAPK in VLC-FFA + p38MAPK-Ⅰ group was significantly decreased (P ＜ 0.05 ),but there was no difference in protein expression ( P ＞ 0.05).(2) The mRNA expression of COX-2 in LC-FFA + DMEM,VLC-FFA +DMEM,LC-FFA + NADPH-Ⅰ,LC-FFA + p38MAPK-Ⅰ,VLC-FFA + NADPH-Ⅰ,VLC-FFA + p38MAPK-Ⅰ group were 3.97 ±0.03,39.08 ±0.63,0.99 ±0.13,0.98 ±0.18,20.93 ±3.70,13.46 ± 2.31 separately,and the protein expressions were 1.32 ± 0.20,1.33 ± 0.25,0.59 ± 0.13,0.58 ± 0.30,0.88 ± 0.18,0.91 ± 0.24.Compared with other groups,mRNA and protein expressions of COX-2 in LC-FFA + DMEM and VLC-FFA + DMEM group were significantly increased ( P ＜ 0.05 ).Compared with LC-FFA + DMEM group,mRNA and protein expressions of COX-2 in LC-FFA + NADPH-Ⅰ and LC-FFA +p38MAPK-Ⅰ group were decreased ( P ＜ 0.05 ).Compared with VLC-FFA + DMEM group,mRNA and protein expressions of COX-2 in VLC-FFA + NADPH-Ⅰ and VLC-FFA + p38MAPK-Ⅰ group were all decreased ( P ＜ 0.05 ).( 3 ) The correlation analysis showed that there were significantly positive correlations between the mRNA and protein expressions of p38MAPK and COX-2 in LC-FFA group ( P ＜ 0.05 ).There were significantly positive correlations in protein expression ( P ＜ 0.05 ),but no conrelation in the mRNA expression between p38MAPK and COX-2 in the F-FFA,SC-FFA,MC-FFA,VLC-FFA groups (P ＞ 0.05).ConclusionsThe oxidative stress and inflammation may exist in trophoblast cells which were stimulated by LC-FFA and VLC-FFA.p38MAPK signal transduction pathway may contributed in this process.

Effects of transient intensive insulin treatment on free fatty acid in newly diagnosed type 2 diabetic patients / 中国医师进修杂志

Objective To study the effect of transient intensive insulin treatment on the serum free fatty acid (FFA) in newly diagnosed type 2 diabetic patients.Methods Sixty-four newly diagnosed type 2 diabetic patients were treated with transient intensive insulin.The fasting plasma glucose (FPG),2 hours post-prandial glucose (2hPG),lipid,fastin insulin (FINS),and serum FFA was examined hefore and after treatment.Results The levels of FPG,2hPG,total cholesterol (TC),triglycerides (TG),low density lipoproteins cholesterol (LDL-C),FFA and HOMA-IR after treatment were (9.68 ± 2.02) mmol/L,(12.77 ± 1.35) mmol/L,(4.26 ± 1.07) mmol/L,(1.52 ± 0.58) mmol/L,(2.50 ±0.75) mmol/L,(435.84 ± 190.94) μmol/L,0.51 ± 0.62,and they decreased obviously compared with those before treatment [(14.66 ± 3.50) mmol/L,(17.43 ±4.89) mmol/L,(5.03 ±0.94) mmol/L,(2.05 ± 1.42) mmol/L,(2.91 ±0.78) mmol/L,(586.68 ±229.45)μmol/L,0.65 ± 0.89](P＜0.05).The level of HOMA-β increased obviously (2.70 ± 0.83 vs.1.74 ± 1.04)(P＜0.05).The increase of HOMA-β and the decrease of HOMA-IR was positively correlated with the decrease of FFA.Conclusion The transient intensive insulin treatment can evidently decrease the level of FFA that can improve beta-cell function and relieve insulin resistance in newly diagnosed type 2 diabetic patients.

Skin Aging and Photoaging Alter Fatty Acids Composition, Including 11,14,17-eicosatrienoic Acid, in the Epidermis of Human Skin

We investigated the alterations of major fatty acid components in epidermis by natural aging and photoaging processes, and by acute ultraviolet (UV) irradiation in human skin. Interestingly, we found that 11,14,17-eicosatrienoic acid (ETA), which is one of the omega-3 polyunsaturated acids, was significantly increased in photoaged human epidermis in vivo and also in the acutely UV-irradiated human skin in vivo, while it was significantly decreased in intrinsically aged human epidermis. The increased ETA content in the epidermis of photoaged human skin and acute UV-irradiated human skin is associated with enhanced expression of human elongase 1 and calcium-independent phophodiesterase A2. We demonstrated that ETA inhibited matrix metalloproteinase (MMP)-1 expression after UV-irradiation, and that inhibition of ETA synthesis using EPTC and NA-TCA, which are elongase inhibitors, increased MMP-1 expression. Therefore, our results suggest that the UV increases the ETA levels, which may have a photoprotective effect in the human skin.

Analysis of the plasma free fatty acid composition in T2DM by high performance liquid chromatography / 中华检验医学杂志

Objective To analyze the plasma free fatty acid (FFA) composition in patients with T2DM. Methods All subjects were from Zhongnan hospital of Wuhan university, and they were divided into three groups: normal control ( n = 94 ), T2DM ( n = 101 ) and T2DM with hyperlipidemia ( n = 77 ). Fasting blood samples were taken from the participants, and plasma FFA were separated using a modified Doles method with the bromoacetophenone, pre-column-derivative. The quantitation of FFA was performed on were (355.63 ± 100. 35) μmol/L, (421.21 ± 200. 83 ) μ mol/L, ( 473.04 ± 213.40 ) μmol/L in healthy controls, T2DM group and T2DM with hyperlipidemia group, respectively. The significant differences were observed among the 3 groups(x2 = 13.08, P <0.01 ). However, there was no significant difference of UFA concentrations among the 3 groups [(206.29± 61.94) μ mol/L, (218.11 ± 110.28) μmol/L and ( 240.94 ± 116.79 ) μmol/L, x2 = 2.17, P > 0.05]. Compared to normal control [( 355.63 ± 100.35 )μmol/L], the FFA concentration[(421.21 ±200.83) μmol/L] in T2DM has significantly increased (x2 =FFA concentrations were higher in T2DM with hyperlipidemia [(473.04 ±213.40) μmol/L] (x2 =27.93,P <0.01 ). The RSD values for intra- and inter-day precision were less than 5%, and the minimal detection limits ranged from 0.05 μmol/L to 0.35 μmol/L The recoveries of high, intermediate and low-level materials were 96.4% -104.8%. Conclusions The total FFA concentration in T2DM has increased, most of which are saturated FFA. The unsaturated FFA has not significantly increased. They seem to be related to the development of T2DM, and might be a new biomarker for clinical monitoring of metabolic disorder of T2DM.

Elevated circulating free fatty acids level causes pancreatic islet β-cell dysfunction via oxidative stress / 中华内分泌代谢杂志

Objective To evaluate the effects of elevated circulating free fatty acids (FFA) level on basal and glucose stimulated insulin secretion (GSIS) of islet β-cell and to explore the pathophysiological link between FFA and impaired β-cell dysfunction. Methods Male SD rats underwent infusions with normal saline (C group), intralipid+heparin (FFA group) and N-acetylcysteine+FFA (NAC group) for 2-4 days. Insulin secretion from pancreatic tissues was evaluated during intravenous glucose tolerance test and isolated pancreas perfasion test at the end of 2 and 4 days infusion. Results After 2 days infusion, the basal insulin secretion from isolated perfused pancreas was increased in FFA group [(55.5±19.4 vs 27.4±6.7) mU/L, P<0.01], but the response to 16.7 mmol/L glucose in isolated perfased pancreas was similar in FFA and C groups. The peak value during GSIS was inhibited by 4 days FFA infusion [(46.8±33.0 vs 214.7±27.4)mIU/L,P<0.05]. GSIS was also decreased in FFA group compared with C group in IVGTr. After interfered with NAC, GSIS was partly recovered [(165.4± 14.8)mIU/L, P<0.01]. Conclusion Elevated circulating FFA levels may contribute to the abnormality of pancreatic islet β-cell through oxidative stress.