Inhibitory effects of apolipoprotein M on inflammatory factors induced by high glucose in human retinal vascular endothelial cells / 中华实验眼科杂志

Objective To observe the expressing changes of apolipoprotein M (ApoM),tumor necrosis factor-α(TNF-α) and monocyte chemoattractant protein-1 (MCP-1) in human retinal vascular endothelial cells (HRECs) under the high glucose culture condition and investigate the inhibitory effects of ApoM overepression on the expressions of TNF-α and MCP-1.Methods HRECs were cultured in DMEM containing 10％ fetal bovine serum and 5.5 mmol/L D-glucose and assigned to 6 groups.The cells in the normal control group were cultured in above culture medium;the cells in the high glucose group were treated using the DMEM with 30 mmol/L D-glucose;ApoM was transfected into the cells using lentiviral vector in the ApoM transfected group;lentiviral vector without ApoM sequence was transfected in the empty vector group;the cells transfected by empty vector were cultured in high glucose culture medium in the empty vector+high glucose group;the cells in the ApoM transfection+high glucose group were treated by ApoM sequence transfection and high glucose incubation.The relative expression of ApoM,TNF-α and MCP-1 mRNA was detected using real-time quantitative PCR,and the relative expression of ApoM protein was evaluated using Western blot assay.Results Compared with the normal control group,the mRNA expression levels of ApoM,TNF-α and MCP-1 in the high glucose group were significantly increased (t=5.517,3.295,2.555;all P＜0.05).HRECs grew well after infected with lentivirus.The relative expression level of ApoM mRNA in the ApoM transfected group was 236.400±39.270,which was significantly higher than 1.000±0.153 in the empty vector group (t=5.995,P＜0.01).An enhanced protein band of ApoM was seen in the ApoM transfected group,and the protein band was absent in the empty vector group.The relative expression band in the ApoM transfected group was 1.000± 0.249 and 2.978 ± 0.285 in the cells cultured with normal culture medium or high glucose culture medium,respectively,with a significant difference between them (t =5.056,P＜0.01).The relative expressions of TNF-α and MCP-1 in the mRNA levels were significantly different among the empty vector group,empty vector+high glucose group,ApoM transfected group and ApoM transfection + high glucose group (F =5.966,P =0.026;F =14.410,P =0.002).Compared with the empty vector+high glucose group,the relative expressions of TNF-α and MCP-1 mRNA were considerably reduced in the ApoM transfection+high glucose group (P=0.017,0.004).Conclusions High glucose environment up-regulates the expression of ApoM,MCP-1 and TNF-α in HRECs.Overexpression of ApoM inhibits the up-regulation of MCP-1 and TNF-α expression induced by high glucose.

The Effects of Short-term Circuit Resistance Training with Supplementation of Milk, Cheese Juice, Whey Protein on Resting Level Apolipoprotein M- HDL Total Cholesterol, Lipid Profile, Gastric Inhibitory Polypeptide and Plasma Estradiol in Untrained Young Men.

Cardiovascular diseases are the most significant factors leading to death. This quasi-experimental study was carried out on 28 non athletic men. Subjects were divided into 4 groups. To the first group water (400 cc) was given, the second group received whey protein supplementation (50g), the third group had cheese juice (50g), and milk was given to the fourth one (400 cc), immediately after the training session. High-density lipoprotein (HDL), its sub fractions HDL2 and HDL3 cholesterol, apolipoprotein M, gastric inhibitory polypeptide and estradiol were evaluated prior to and after training for 12 sessions, three sessions in each week. The differences were not significant for gastric inhibitory polypeptide (GIP) and estradiol in the fourth group after training in comparison with the first phase. A significant decrease in apolipoprotein M- HDL total cholesterol on resting level were observed in all four groups and in HDL3 cholesterol with cheese juice and also whey protein in comparison with prior phase (P-values were 0.02, 0.001, 0.000, 0.015, respectively) (0.01, 0.01). A significant increase in HDL2 cholesterol level was observed in the groups with cheese juice and whey protein in comparison with their prior phases (P-values were 0.02, 0.03, respectively). This study showed that circuit resistance training has positive effects on the lipid profile changes in untrained young men.

Identification and application of sphingosine-1-phosphate receptor of human umbilical vein endothelial cells / 天津医药

Objective To identify the expression of sphingosine-1-phosphate receptor (S1PR) subtypes, C-myc and His tag proteins of human umbilical vein endothelial fusion cell line, EA.hy926 and human umbilical vein endothelial cells (HUVEC), CRL-1730 for studying the function of apolipoprotein M (ApoM)-S1P axis. Methods Two kinds of cells (EA. hy926 and CRL-1730) were cultured to reach the density of 60%-70% in vitro. Immunofluorescence technique was em?ployed to investigate the expressions of coagulation factorⅧ(FⅧ), ApoM, S1PR1-S1PR5, C-myc and His tag proteins. Re?sults (1) Two kinds of cells both expressed FⅧand ApoM. FⅧpresented scattered particle distribution in CRL-1730, while uniform distribution in EA.hy926. However, ApoM was strongly expressed and widely distributed in cytoplasm of two kinds of cells. (2) S1PR1-3 can be detected on their membrane other than S1PR4 and S1PR5. S1PR1 was highly expressed but S1PR2 and S1PR3 were in a low level expression. (3) Two kinds of cells both expressed C-myc and His tag proteins in cytoplasm. Conclusion Two kinds of cells have the properties of endothelial cells and can express FⅧ, ApoM, C-myc and His tag proteins. It is not suitable for choosing C-myc and/or His tag–conjugated recombinant ApoM to study the fuction of ApoM-S1P axis with these two kinds of cells.

Value of serum APOM concentration for diagnosis of bacterial pneumonia and judgment of disease severity / 国际检验医学杂志

Objective To investigate the value of serum apolipoprotein M (APOM)concentration in the diagnosis of bacterial pneumonia and its disease severity evaluation.Methods The serum APOM concentration was examined in 250 cases of pneumonia and 50 healthy controls by using the double antibody sandwich ELISA.The ROC curve was used to evaluate the diagnostic value of APOM on bacterial pneumonia and the Bayes discriminant analysis was adopted to construct the discriminant equation based on APOM concentration for evaluating its discriminant significance to the disease severity.Results The serum APOM concentration in the bacterial pneumonia group was (28.9±10.5)μg/mL,which was significantly higher than (19.4±8.93)μg/mL in viral pneu-monia group,(16.4±5.62)μg/mL in the other pneumonia group and(18.1 ±6.15)μg/mL in the healthy controls,the differences had statistical significance(P <0.01).The area under ROC curve for APOM in diagnosing bacterial pneumonia could reach 0.843, showing the good sensitivity and specificity.Moreover,the serum APOM concentration was positively correlated with the disease severity of bacterial pneumonia.The discriminant equation based on APOM concentration as well as sex,age,systolic pressure,body temperature and heart rate could better conduct the classification on the disease severity of bacterial pneumonia and its accuracy could reach to 86.7%.Conclusion The serum APOM concentration has an important clinical value for the diagnosis of bacterial pneumonia and its disease severity evaluation.

Changes of serum apolipoprotein M level in women during normal and gestational diabetes mellitus / 中华内分泌代谢杂志

Objective To investigate the changes of serum apolipoprotein M (ApoM) level in cord blood and pregnant women during different gestational periods and to explore the relationship with the occurance and development of gestational diabetes mellitus (GDM).Methods Three hundred and eight pregnant women performing regular prenatal examination were registered in Changzhou Women and Children Health Hospital from June,2011 to April,2013,including 99 normal pregnant women during the second trimester,122 during the third trimester,and 87 pregnant women with GDM.At the same time,100 samples of cord blood of normal neonates were collected.The levels of serum ApoM and other lipids were measured.Results (1) Serum ApoM levels in pregnant women during the second trimester,during the third trimester,and in the cord blood of normal neonate were (0.036-± 0.013) g/L,(0.023 ± 0.008) g/L,and (0.010 ± 0.004) g/L,respectively.(2) The serum ApoM level of GDM group was lower than that of control [(0.029 ± 0.010 vs 0.036 ± 0.013) g/L,P＜0.05].Multivariate non-conditional logistic regression analysis showed that there was statistically significant difference in ApoM levels between GDM and control groups after adjusting for potential confounders (Z =-3.62,P＜0.05).Conclusions The level of serum ApoM in pregnant women was increased at first and then decreased.The serum ApoM level in the neonatal cord blood was significantly lower than that in pregnant women.The level of serum ApoM was lowered with raised blood glucose level in women with GDM,suggesting that ApoM might play a role in the pathological processes of GDM.

Effects of phospholipid transfer protein overexpression on content of sphingosine-1-phosphate in mouse lipoprotein / 中国病理生理杂志

[ABSTRACT]AIM:Toinvestigatetheinteractionandthemechanismofsphingosine-1-phosphate(S1P)and phospholipid transfer protein (PLTP) in lipoprotein.METHODS:The S1P content in the plasma and lipoprotein from 10-week-old PLTP transgenic (PLTP-Tg) mice and wild-type (WT) mice (n=8 each) was assayed.The transport of S1P by PLTP was determined by S1P transfer assay.The content of specific S1P carrier, apolipoprotein M, was detected by West-ern blotting.RESULTS:Plasma S1P contents were decreased by 21.1%in PLTP-Tg mice compared with WT mice.S1P content in high-density lipoprotein ( HDL) fraction ( HDL-S1P) from PLTP-Tg mice was decreased by 35.1% compared with WT mice, whereas the S1P in low-density lipoprotein (LDL) fraction (LDL-S1P) was increased by 127.4%.The re-sults of S1P transfer assay indicated that PLTP facilitated S 1P transport from erythrocyte to recombinant liposome at 37℃in D-Hanks buffer solution .The plasma content of apolipoprotein M was not changed in PLTP-Tg mice compared with WT mice.CONCLUSION:PLTP is a key factor to maintain plasma HDL-S1P under physical condition .Overexpression of PLTP decreases the HDL-S1P but increases LDL-S1P.The mechanism might be related to the capability of PLTP on trans-ferring S1P from erythrocyte to lipoprotein.

Expression of apolipoprotein M in renal graft in rats and its roles and action mechanism in acute rejection / 中华器官移植杂志

Objective To investigate the expression of apolipoprotein M in rats after renal transplantation and its role and action mechanism in acute rejection (AR).Methods The kidney transplantation model in rats were established.Male SD and Wistar rats were used as donors,and Wistar rats as recipients.Three groups were designated:control group (syngeneic transplantation,n =24,recipients were treated with normal saline i.p.); AR group (allogeneic transplantation,n =24,recipients were treated with normal saline i.p.); PDTC group [allogeneic transplantation,n =24,recipients were treated with NF-κB inhibitor-pyrrolidine dithiocarbamat (PDTC) i.p.].The renal grafts were drawn at day 1,3,5 and 7 post-transplantation,and the expression levels of NF-κB P65 and apoM protein were detected by using Western blotting,and those of apoM,perforin and granzyme B mRNA were by using real-time PCR.The correlation of apoM and NF-κB,apoM and perforin,and apoM and granzyme B was respectively analyzed.Results As compared with control group,the expression levels of apoM,perforin and granzyme B mRNA in AR and PDTC groups were dramatically up-regulated at each time point (P＜0.01),and those in PDTC group were significantly lower than those in AR group (P＜0.01).The expression levels of apoM and NF-κB protein in AR group were both distinctly higher than those in control group (P＜0.01),and those in PDTC group were markedly lower than those in AR group (P＜0.01).A significantly positive correlation was found between the expression of apoM and NF-κB protein (r =0.469,P＜0.05).And the expression of apoM mRNA was positively correlated to perforin and granzyme B mRNA (r =0.731,P＜0.01 ; r =0.514,P＜0.05).Conclusion The expression of apoM is obviously up-regulated in renal grafts of rats,which may take part in the pathogenesis of AR via NF-κB.

Estrogen increases apolipoprotein M through estrogen receptor / 中华内分泌代谢杂志

Objective To explore the effects of estrogen on apolipoprotein M ( apoM ).Methods ApoM mRNA was assayed in HepG2 cells by RT-PCR after incubation of estrogen with or without estrogen receptor antagonist at different concentrations and durations.SD female rats were divided into five groups:OVX group,Sham group,OVX+ EB group,normal group and normal + EB group.From a week of being operated,the rats were injected subcutaneously estradiol beuzoate or vehicle.After 12-hrs fasting,serum levels of triglycerides (TG),LDL-cholesterol,HDL-cholesterol,total cholesterol ( TC ) at months 1,2 and 3 after operation were measured.The expression of apoM in rats was detected by using real time RT-PCR and Western blot.Results Estrogen increased mRNA levels of apoM and apoAI in the HepG2 cells with a dose- and time-dependent manner,which could be abolished by addition of estrogen receptor antagonist.Serum apoM,TG,TC,HDL and LDL levels were significantly increased in the ovariectomized or normal rats which received estrogen treatment than those in OVX or normal group rats at month 1 after treatments.Conclusions Estrogen upregulates apoM expression via its receptor.

Correlation of apolipoprotein M with severity of coronary artery stenosis in patients with coronary heart disease / 中华全科医师杂志

One hundred and thirty seven pafients diagnosed by coronary angiography were recruited in the study and divided into coronary heart disease group(CAD)and control group.Relationship between serum apolipoprotein M levels and the severity of stenosis of coronary arteries was analyzed.In covariate analysis,serum apolipoprotein M levels were significantly lower in CAD group than those in control group adjusted for sex,age,body mass index(BMI)(P＜0.05).By stepwise muhiple regression,serum levels of apolipoprotein M were correlated positively with BMI(r=0.65,P＜0.01),and correlated negatively with total cholesterol(TC)(r=-0.53,P＜0.01)and low density lipoproteins(LDL-C)(r=-0.42,P＜0.01).Serum levels of apolipoprotein M were correlated negatively with the score of coronary stenosis(r=-0.62,P＜0.01).The results suggest that apolipoprotein M might be related to the development of coronary artery disease.