Analysis of serum metabolic profiling of preeclampsia pregnancy / 中华检验医学杂志

Objectives This research explored the characteristics of changes in the serum metabolic profile of preeclampsia pregnancy(PE) to establish the disease distinguish model and screen characteristic metabolic markers with potential diagnostic value for preeclampsia.Methods From August 2014 to January 2016,samples in three groups were collected at Tianjin Third Central Hospital.Thirty-one clinically diagnosis patients with preeclampsia,25 normal pregnancy women and 29 healthy volunteers of childbearing age were enrolled.Ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) was used to analyze serum metabolites of PE group (31 patients with preeclampsia),P group (25 normal pregnancy women) and Normal group (29 healthy volunteers of childbearing age).Nonparametric test analyzes were used to analyze the data and find the specific metabolites.Results This research established the principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) disease distinguish model for PE group,P group and Normal group.To distinguish PE group,P group and Normal group,15 characteristic metabolites were identified.Eight kinds of glycerol phospholipid (including 7 kinds of hemolysis phosphatidyl choline and 1 kind of lysophospholipids acid) and 1 kind of sphingomyelin in PE group were higher than that of normal pregnancy group.The difference had statistically significant(Z of the metabolites were 2.32,3.34,3.21,2.60,2.22,3.40,3.58,5.84,2.70 respectively,all P<0.05).1,25-Dihydroxyvitamin D3-26,23-lactone and 24-Oxo-1alpha,23,25-trihydroxyvitamin D3 in PE group were higher than that of P group and Normal group,which had a statistics difference (Z of the metabolites were 2.01,3.89,3.26,2.34 respectively,all P<0.05).Conclusions Metabolomics distinguish model has a good ability to distinguish PE group,P group and Normal group.Serum characteristic metabolites can successfully reflect the status of fat,calcium and phosphorus metabolism of preeclampsia patients and provide high value for prediction,diagnosis and treatment.

RAGE/NF-κB signal pathway in mediating lysophosphatidylcholine-induced TGF-β1 expression in human retinal endothelial cells / 重庆医学

Objective To investigate the role of receptor for advanced glycation end products (RAGE)/NF-κB signaling pathway in mediating lysophosphatidylcholine (LPC)-induced TGF-β1 expression in human retinal endothelial progenitor cells (HEPCs).Methods Human retinal endothelial cells (HERCs)were transfected with siRNA for RAGE siRNA or added NF-κB in-hibitor pyrrolidine dithiocarbamate (PDTC)in the presence or absence of LPC,the expressions of TGF-β1 and RAGE genes were analyzed by qPCR and Western blot.Results LPC could increase the expression of RAGE and TGF-β1 gene in HERCs.The RAGE gene after silence could significantly decrease the expression of LPC-induced RAGE and TGF-β1 .Adding NF-κB inhibitor PDTC significantly reduced LPC-induced RAGE and TGF-β1 expression in HERCs.Conclusion RAGE/NF-κB signaling pathway plays an important role in mediating LPC induced TGF-β1 gene expression in HERCs.

Research progress of lysophosphatidylcholines for liver diseases / 中国药理学通报

Lysophosphatidylcholines belong to a group of lipid components which have a variety of physiological functions. LPCs are known to be linked to metabolic disorders and cardio-vascular diseases,including diabetes,atherosclerosis and dyslip-idemia.LPCs are actively metabolized in liver,which is closely related with liver diseases and hepatotoxicity.The role of LPCs in liver diseases and hepatotoxicities has been extensively investi-gated recently.This review focuses on lysophosphatidylcholines as a biomarker for liver diseases,such as hepatic carcinoma, cholestasis,cirrhosis,hepatitis,and chemical hepatotoxicities, trying to lay a basis for investigation and therapeutics of liver dis-eases.

Lysophosphatidylcholine, Oxidized Low-Density Lipoprotein and Cardiovascular Disease in Korean Hemodialysis Patients: Analysis at 5 Years of Follow-up

Although oxidized low-density lipoprotein (LDL) and lysophosphatidylcholine (LPC) have been proposed as important mediators of the atherosclerosis, the long-term contribution to the risk of cardiovascular disease (CVD) in hemodialysis patients has not been evaluated. This study investigated the relation between oxidized LDL and LPC levels with long term risk of CVD. Plasma oxidized LDL and LPC levels were determined in 69 Korean hemodialysis patients as a prospective observational study for 5 yr. During the observation period, 18 cardiovascular events (26.1%) occurred including 6 deaths among the hemodialysis patients. The low LPC level group ( 254 microM/L) (P = 0.01). However, serum levels of oxidized LDL were not significantly different between groups with and without CVD. In adjusted Cox analysis, previous CVD, (hazard ratio [HR], 5.68; 95% confidence interval [CI], 1.94-16.63, P = 0.002) and low LPC level (HR, 3.45; 95% CI, 1.04-11.42, P = 0.04) were significant independent risk factors for development of CVD. It is suggested that low LPC, but not oxidized LDL, is associated with increased risk of CVD among a group of Korean hemodialysis patients.

Cytotoxic Effects of Lysophosphatidylcholine on Vascular Smooth Muscle Cells / 대한폐경학회지

OBJECTIVES: To investigate the cytotoxic effects of lysophosphatidylcholine (lysoPC), an active component of oxidized low-density lipoproteins (LDL), on vascular smooth muscle cells (VSMCs). METHODS: VSMCs were derived from rat aorta. Cell death was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, lactic dehydrogenase (LDH) assay, and DNA fragmentation assay. Apoptosis was quantified by propidium iodide staining and fluorescent activated cell sorting (FACS) analysis, and intracellular free radical production was determined using 2',7'-dichlorofluorescin diacetate (DCF-DA). In addition, the changes in caspases, bcl-2 and bax proteins were evaluated by western blot analysis. RESULTS: LysoPC over 25 microM induced more than 50% of the cell death at 10 hours on MTT assay with no change in the level of LDH. The DNA ladder pattern showed that cell death induced by lysoPC was caused by apoptosis, which was associated with increased free radical production. Vitamin E, a potent antioxidant and caffeic acid phenylethyl ester (CAPE), an inhibitor of nuclear factor-kappaB (NF-kappaB), blocked apoptosis. The casepase-3 precursor decreased and the active form of caspase-8 increased. Total bcl-2 and bax proteins did not change with lysoPC treatment, but translocation of bax from cytosole to the mitochondria membrane was observed. CONCLUSION: LysoPC induces apoptosis in VSMCs via an oxidant mechanism, dependent on NF-kappaB.

Impact of Lysophosphatidylcholine on the Plasminogen Activator System in Cultured Vascular Smooth Muscle Cells

The balance between tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor type 1 (PAI-1) regulates fibrinolysis. PAI-1 expression increases in atherosclerotic arteries and vascular smooth muscle cells (VSMCs) are one of major constituents of atheroma. We investigated the impact of lysophosphatidylcholine (lysoPC), an active component of oxidized low-density lipoprotein, on the plasminogen activator system of the rat VSMCs. The lysoPC stimulated the protein and gene expressions of PAI-1 but did not affect the protein expression of t-PA. Fibrin overlay zymography revealed that lysoPC increased the activity of PAI-1 in the conditioned media, while concurrently decreasing that of free t-PA. Vitamin E inhibited the lysoPC-induced PAI-1 expression. Further, lysoPC increased the intracellular reactive oxygen species (ROS) formation. Caffeic acid phenethyl ester, an inhibitor of NF-kappaB, blocked this lysoPC effect. Indeed, lysoPC induced the NF-kappaB-mediated transcriptional activity as measured by luciferase reporter assay. In addition, genistein, an inhibitor of protein-tyrosine kinase (PTK), diminished the lysoPC effect, while 7,12-dimethylbenz[a]anthracene, a stimulator of PTK, stimulated PAI-1 production. In conclusion, lysoPC does not affect t-PA expression but induces PAI-1 expression in the VSMC by mediating NF-kappaB and the genistein-sensitive PTK signaling pathways via oxidative stress. Importantly, lysoPC stimulates the enzyme activity of PAI-1 and suppresses that of t-PA.

The effects of Tong Xin-Luo on human umbilical vein endothelial cells impaired by lysophosphatidylcholine / 中国医师杂志

Objective To investigate the effects of Tong Xin-Luo on cultured human umbilical vein endothelial cells (HUVECs) impaired by Lysophosphatidylcholine.Methods The herbage-contained serum of TXL was prepared,HUVECs were cultured in vitro.The study was designated to 4 group:normal control,LPC group,TXL group,and TXL + LPC intervened group.The cell function was determined by cell morphology and MTT colorimetric assay.Results Compaired with normal control group (0.380 ±0.023 ),LPC ( 0.320 ± 0.024 ) could significantly decrease the cells activity,promote cells death ( P ＜0.05 ).After TXL intervened(0.424 ±0.034),cells activity was significantly increased,cells death was d significantly decreased( P ＜0.05 ).Conclusions Tong Xin-Luo could protect human umbilical vein endothelial cells function by against the LPC-induced damage.

Advances in the study of arrhythmogenic mechanisms / 国际药学研究杂志

Arrhythmia is a common complication of cardiovascular diseases and a risk factor for human health. Especially, ventricular tachycardia and ventricular fibrillation may not only exacerbate original heart diseases, but also cause cardiac sudden death which has been an mportant death reason in China. However, anti-arrhythmic drugs nowadays cannot effectively treat these arrhythmias, with an efficiency of only 30%-60%, which indicates that our knowledge about arrhythmias is limited. Hence, to explore the potential mechanism, look for novel targets, and develop drugs with multiple-channel action are the focus of the research direction. Recent studies displayed that the atrial-specific potassium channels such as IKur and IKAch were involved in atrial fibrillation, which provided a prospective target for atrial fibrillation treatment. Calcium leak, gap junction protein and autoantibody against ICaL channel were shown to participate in arrhythmogenesis. These findings provided a theoretical basis for the development of more effective anti-arrhythmic drugs. Remarkably, as a kind of mportant RNA regulating gene expression, microRNA (miRNA) was shown to possess anti-arrhythmic activities which may prevent cardiac sudden death. miR-1, miR-133 and miR-590 regulated the arrhythmia in various types of animal models. Because of the multiple-gene regulation actions of miRNA, it has the potential to be developed as novel anti-arrhythmic target.

Lysophosphatidylcholine suppresses apoptosis and induces neurite outgrowth in PC12 cells through activation of phospholipase D2

Lysophosphatidylcholine (LPC) is a bioactive lipid generated by phospholipase A2-mediated hydrolysis of phosphatidylcholine. In the present study, we demonstrate that LPC stimulates phospholipase D2 (PLD2) activity in rat pheochromocytoma PC12 cells. Serum deprivation induced cell death of PC12 cells, as demonstrated by decreased viability, DNA fragmentation, and increased sub-G1 fraction of cell cycle. LPC treatment protected PC12 cells partially from the cell death and induced neurite outgrowth of the cells. Overexpression of PLD2 drastically enhanced the LPC-induced inhibition of apoptosis and neuritogenesis. Pretreatment of the cells with 1-butanol, a PLD inhibitor, completely abrogated the LPC-induced inhibition of apoptosis and neurite outgrowth in PC12 cells overexpressing PLD2. These results indicate that LPC possesses the neurotrophic effects, such as anti-apoptosis and neurite outgrowth, through activation of PLD2.

Development of ligand-dependent regulatory system and its application to gene therapy of insulin-dependent diabetes mellitus

To develop an inducible expression system, the enhanced artificial nuclear receptors and target reporters were constructed. Artificial nuclear receptors were generated by fusing three domains, consisting of DNA-binding domain (DBD) of GAL4, ligand binding domain (LBD) of progesterone or estrogen receptor, and activation domain (AD) of VP16, sterol regulatory element binding protein (SREBP)-1a, or SREBP-2. The activation domain of SREBP-1a showed most potent transcriptional activity. The maximal level of target reporter gene expression was extremely elevated by the usage of ATP citrate-lyase (ACL) minimal promoter -60/+67 in place of artificial TATA promoter, while the SV40 enhancer severely increased the basal transcription in the absence of ligand. The induction system, developed in the present study, was applied to cell therapy, resulting in successful induction of single-chain insulin analogue (SIA) gene expression to correct the hyperglycemia in diabetic animals. By means of subcutaneous cell therapy, the SIA gene expression rapidly occurred after the local topical application of ligand. These results suggest that our system represents a powerful tool for transcriptional regulation of target gene that can be used for diverse applications, ranging from basic research to gene therapy.

The Effect of Naringin on the Lysophosphatidylcholine-induced Proliferation of Vascular Smooth Muscle Cells

BACKGROUND AND OBJECTIVES: Lysophosphatidylcholine (lysoPC), an atherogenic lysophospholipid, is known to induce the proliferation of vascular smooth muscle cells (VSMCs). Naringin is a flavonoid in grapes and grapefruits and has anti-inflammatory as well as antioxidative effects. We investigated whether naringin could protect VSMCs from the effect of lysoPC. Additionally, we investigated the changes of nuclear translocation of nuclear factor-kappa B (NF-kappa B). MATERIALS AND METHODS: VSMCs were prepared from the aorta of Sprague-Dawley rats. Near-confluent VSMCs were preincubated in media containing 0, 10, 100 micrometer naringin, and incubated with 0, 10, 20, 100 micrometer lysoPC. The degree of proliferation of VSMCs was evaluated with [H3]-thymidine incorporation and MTT assay. The changes of nuclear translocation of NF-kappa B in VSMCs were investigated with EMSA. RESULTS: LysoPC promoted the growth of VSMCs, whereas naringin inhibited the proliferative effect of lysoPC on VSMCs. MTT assay showed a 63+/-24% and 89+/-17% increase of cellular growth in the 10 and 20 micrometer lysoPC groups, respectively, as compared with the control group with media only (p<0.01). [H3]-thymidine incorporation assay also showed 61+/-25% and 92+/-25% increase in the 10 and 20 micrometer lysoPC groups, respectively (p<0.01). However, the growth of VSMCs was suppressed in the 100 micrometer lysoPC group (p<0.01). Naringin inhibited the proliferative effects of lysoPC on VSMCs by 34+/-5% (MTT assay) and 35+/-5% ([H3]-thymidine incorporation assay) in the 100 micrometer naringin group (p=0.01). The nuclear translocation of NF-kappa B was stimulated by lysoPC and suppressed by naringin. CONCLUSION: LysoPC promoted the growth of VSMCs, whereas naringin inhibited the proliferative effect of lysoPC on VSMCs. These effects of lysoPC and naringin are associated with the regulation of nuclear translocation of NF-kappa B.

The effect of lysophosphatidyl choline on the expression of scavenger receptor class B type Ⅰ in L-20 cells and the impact of simvastatin / 中国医师杂志

Objective To observe the impact of lysophosphatidyl choline(LPC) on the expression of scavenger receptor class B type Ⅰ(SRBI) in L-20 cells and the impact of simvastatin.Methods Cultured hepatocytes were randomly assigned to normal group,LPC-damaged group and simvastatin group.The changes of mRNA and protein expression of SRBI in hepatic tissue were assayed by RT-PCR,immunocytochemistry and Western blot,respectively.Results The expression of SRBI was higher in LPC-damaged hepatocytes than that in normal cells at both mRNA and protein level(P

Effects of oxidized low density lipoprotein and lysophosphatidylcholine on cholesterol efflux from mouse peritoneal macrophage foam cells / 中国病理生理杂志

AIM: To explore the effects of oxidized low density lipoprotein (OxLDL) and one of its component— lysophosphatidylcholine (LPC) on cholesterol efflux from mouse macrophage foam cells. METHODS: (1) Cholesterol efflux induced by apoAI from mouse peritoneal macrophage foam cells loaded with OxLDL or acylated LDL(AcLDL) was measured. (2) Cholesterol efflux induced by LPC and apoAI from macrophage foam cells separated from normal or apoE gene deficient (E 0) mouse loaded with AcLDL were measured. RESULTS: (1) When the macrophage foam cells were incubated with apoAI, cholesterol efflux from AcLDL-induced macrophage foam cells increased significantly compared to that of OxLDL-induced macrophage foam cells. (2) LPC promoted cholesterol efflux from macrophage foam cells in relation to both dosage and time. When LPC was incubated with E 0 mouse macrophage foam cells, the released cholesterol mass was significantly lower than that of normal mouse macrophage foam cells. It was also found that cholesterol efflux induced by apoAI normally occurred in E 0 mouse macrophage foam cells. CONCLUSIONS: (1) OxLDL accumulated cholesterol in macrophages and impair cholesterol efflux. (2) LPC induced cholesterol efflux from macrophage foam cells, which may occur via apoE pathway.

Effect of lysophosphatidylcholine on cholesterol efflux from macrophage foam cells / 中国病理生理杂志

AIM: To explore the effect of lysophosphatidylcholine(LPC) on cholesterol efflux from macrophage foam cells, and to offer experimental evidence for research on prevention and treatment of atherosclerosis.METHODS: Peritoneal macrophages and LDL were seperated from mice and serum of healthy volunteers, respectively. The foam cells were derived from macrophages in the presence of Acylated LDL (AcLDL). Cholesterol efflux from cells and LDH activity were measured by enzymetic fluorometry and LDH kit, respectively.RESULTS: After incubated with LPC for 24 hours, cholesterol efflux from macrophage foam cells increased significantly compared to control, and cellular cholesterol was lower than that in control group. At the same time, medium LDH activity of LPC group was not increased obviously. CONCLUSION: Within the dosage of 10-80 ?mol/L, LPC can promote cholesterol efflux from macrophage foam cells in a dose-dependent manner, and this effect has nothing to do with cytotoxity of LPC.