Inhibitory effects of LDL-ACM complex on subcutaneous implanted tumors in nude mice()

AIM: In order to evaluate the applicable value of LDL as a targeted vehicle for chemotherapeutic agents, we investigated and compared the inhibitory effects of LDL-ACM complex and free ACM on nude mice's subcutaneous implanted tumors derived from gastric cancer cell lines, SGC-7901 and NKM-45. METHODS: LDL-ACM complex was prepared and the tumor model of nude mice was established by subcutaneous implantation of SGC-7901 and NKM-45. Then, the groups of nude mice developed subcutaneous implanted tumors were received either LDL-ACM complex or free ACM. Subsequently, the tumor size, weight and leukemia cell counts were measured and the rates of tumor-inhibition and the survival were compared among the groups. RESULTS: The inhibitory effects of LDL-ACM complex on the tumors, especially on SGC-7901 implanted tumors were much more obvious than that of free ACM. It was also indicated that the action of LDL-ACM complex was mediated by LDL receptor. CONCLUSION: These results showed that LDL-ACM complex had significant inhibitory effects on the implanted tumors and the effect might be mediated by LDL receptor.     

Effects of oxidized low density lipoprotein and lysophosphatidylcholine on cholesterol efflux from mouse peritoneal macrophage foam cells

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 (E⁰) 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 E⁰ 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 E⁰ 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.     

Effects of oxidized high-density lipoprotein on membrane fluidity and the expression of lymphocyte function associated antigen-1 of monocyte

AIM: To investigate the effects of oxidized high-density lipoprotein (oxHDL) on membrane fluidity and the expression of lymphocyte function associated antigen(LFA-1) of monocyte. METHODS: The membrane fluidity of THP-1 cells was assayed by fluorescence anisotropy with DPH (1,6-dipheny-1,3,5-hexatriene), a fluorescent probe; The LFA-1 expression on THP-1 cells were assayed by flow cytometry with indirect immunofluorescence.RESULTS: The membrane fluidity of THP-1 cells was reduced by 45% and 52% respectively (P＜0.01) after incubation with 100 μg protein/mL oxHDL and oxLDL for 20 h; oxHDL could stimulated the expression of LFA-1 on THP-1 cells, the positive cell rate and mean fluoresence intensity (MFI) increased by 39% and 45% respectively versus control group (P＜0.01) after incubation with cells for 24 h. CONCLUSIONS: By increasing the expression of LFA-1 on monocyte, oxHDL can promote the monocyte-endothelium adhesion; oxHDL also can reduce the membrane fluidity of monocyte, that will limit the returning of monocytes from subendothelium back to the circulation. This suggested that oxHDL may play an important role in atherogenesis.     

Effects of a pentanucleotide repeat polymorphism of the apolipoprotein (a)gene on serum levels of lipoproteins in healthy Han population

AIM:To understand the relationship between pentanucleotide repeat(PNR) polymorphism of the apolipoprotein(a) gene and lipoprotein natures of normal Han population. METHODS:The serum levels of TG, TC, LDL-C,HDL-C, apo AI, apo B and Lp(a) were measured and the polymorphism of the apo(a) PNR was studied by using PCR-SSCP in 153 random normal individuals in Hubei Han population respectively. RESULTS:The relative frequencies of apo(a) PNR allele were significantly different from western population. The apo(a) gene which copy number of PNR is 5 was associated with high Lp(a) levels. No marked differences in the levels of TG, TC,LDL-C, HDL-C, apo AI and apo B were found among the various genotype groups of apo(a) PNR in Hubei Han. CONCLUSION:The data of lipids and PNR polymorphism of the apo(a) gene from healthy Hubei Han were obtained. The apo(a) allele with 5(TTTTA)-repeats may be related to high serum Lp(a) levels in the Hubei Han population.     

Effects of oxidized low density lipoprotein and vitamin E on the levels of IL-6,IL-8 and TNF-α in cultured human umbilical vein endothelial cells

AIM:To investigate effects of OX-LDL and VitE on the levels of IL-6,IL-8 and TNF-α in human umbilical vein endothelial cells(HUVEC).METHODS: Human umbilical vein endothelial cells were obtained by in vitro culture. HUVEC treated with or without Vit E was incubated with OX-LDL, and the levels of IL-6, IL-8 and TNF-α were determined by enzyme-linked immunosorbent assy technique. RESULTS:50 μg/L,100 μg/L, 200 μg/L OX-LDL induced the release of IL-6,IL-8 and TNF-α by HUVEC in a dose-dependent manner. Compared with the control group , the levels of IL-6 and IL-8 were significantly increased at 6-12 h of stimulation with OX-LDL . Maximal levels of IL-6 and IL-8 occurred after 24-36 h, reaching a plateau maintained for at least 48 h. TNF-α rose after 2-6 h in HUVEC, and reached a maximum after 12 h. In contrast to IL-6 and IL-8, TNF-α declined after 48 h. However, when VitE (50 mg/L,100 mg/L,200 mg/L)was added, it can significant inhibited the release of IL-6, IL-8 and TNF-α in a dose-dependent manner, and after 48 h these cytokines have no diference between OX-LDL+VitE groups and OX-LDL groups. CONCLUSION: OX-LDL can obviously stimulate the production of IL-6,IL-8 and TNF-α in vascular endothelial cells, which can significantly be inhibited by VitE in a short time.     

Molecular mechanisms of pathogenicity of Mycoplasma mycoides subsp. mycoides SC

Mycoplasma mycoides subsp. mycoides SC, the aetiological agent of contagious bovine pleuropneumonia (CBPP), is considered the most pathogenic of the Mycoplasma species. Its virulence is probably the result of a coordinated action of various components of an antigenically and functionally dynamic surface architecture. The different virulence attributes allow the pathogen to evade the host's immune defence, adhere tightly to the host cell surface, persist and disseminate in the host causing mycoplasmaemia, efficiently import energetically valuable nutrients present in the environment, and release and simultaneously translocate toxic metabolic pathway products to the host cell where they cause cytotoxic effects that are known to induce inflammatory processes and disease. This strategy enables the mycoplasma to exploit the minimal genetic information in its small genome, not only to fulfil the basic functions for its replication but also to damage host cells in intimate proximity thereby acquiring the necessary bio-molecules, such as amino acids and nucleic acid precursors, for its own biosynthesis and survival.     

Effect of inclusion of cashew globulin (Anacardium occidentale) to a casein diet on lipid parameters in rats

The effect of inclusion of cashew globulin to a casein diet on lipid metabolism was studied in rats fed diets with two levels of cashew globulin meal. Inclusion of cashew globulin to a casein diet produced lower levels of total cholesterol, triacylglycerol and phospholipids in the serum and tissues and lower levels of serum lipoprotein cholesterol. There was decreased cholesterogenesis in the liver as evidenced by decreased activity of HMG CoA reductase and decreased release of lipoproteins into circulation. Rats fed cashew globulin along with casein also showed higher activity of LPL in the heart and adipose tissue and higher activity of LCAT. Increased hepatic diversion of cholesterol to bile acid synthesis and increased excretion of bile acids and sterols were also observed in these groups. Activity of glucose-6-phosphate dehydrogenase and malic enzyme was decreased in rats fed cashew globulin along with casein. This study demonstrates that cashew globulins included in the diet of rats are able to alter lipid metabolism which results in lower levels of lipid parameters in the serum and tissues.     

Oxidized low density lipoprotein induces the expression of macrophage migration inhibitory factor

AIM: To examine expression of macrophage migration inhibitroy factor (MIF) gene and protein in macrophages induced by oxidized low density lipoprotein (ox-LDL). METHODS: Macrophages were incubated with ox-LDL at the concentration of 150 mg/L for time course (0-36 h) and with ox-LDL at the different concentrations (0-300 mg/L) for 24 h, expression of MIF mRNA and protein were detected by RT-PCR and ELISA. RESULTS: The results showed that ox-LDL increased MIF gene and protein expression in macrophages in a dose and time-dependent manner. After the exposure of macrophage to ox-LDL, the expression of MIF mRNA level increased consistently with protein. CONCLUSION: MIF may play an important role in atherosclerosis.     

Hemocyanin lipid uptake in Polybetes pythagoricus is altered by fenitrothion

The spider very high density lipoprotein (VHDL), which contains hemocyanin as the major apoprotein, transports most of the circulating lipids. In this work, the effect of the pesticide fenitrothion (FS) on the ability of VHDL-apoproteins to uptake different lipids was investigated. For this, VHDL was delipidated using Triton X-100 and recombined with different radiolabeled lipids in the presence or the absence of FS. The oligomeric structural integrity was maintained after delipidation as shown by non-denaturating PAGE. In the presence of the insecticide, palmitic acid uptake decreased by 28.2 and 62.4% after treating the apolipoprotein with 10 and 20 ppm FS, respectively. Decreases in the uptake of cholesterol, triolein, and phosphatidylcholine caused by FS were 29, 23, and 31% using 10 ppm, and 40, 44, and 29% using 20 ppm FS, respectively. Fluorescence measurements with the hydrophobic probes diphenylhexatriene (DPH) and diphenylhexatrienyl-propionic acid (DPH-PA) indicate that FS induces a red shift, decreases the intensity and increases the anisotropy of the emission of these probes in the VHDL. These results indicate that insecticide binding to the lipoprotein enhances the environment polarity and restricts the mobility of these probes at their binding site. These changes at the hydrophobic VHDL binding sites could lead to the decreased affinity for lipids and hydrophobic ligands. It is inferred that FS could alter the normal lipid exchange between this lipoprotein and tissues.