Stability of serum high-density lipoprotein-microRNAs for preanalytical conditions
Hiroaki IshikawaHiroya YamadaNao TaromaruKanako KondoAyuri NaguraMirai YamazakiYoshitaka AndoEiji MunetsunaKoji SuzukiKoji OhashiRyoji Teradaira
20
Citation
30
Reference
10
Related Paper
Citation Trend
Abstract:
Background Recently, several studies have shown that microRNAs are present in high-density lipoprotein, and high-density lipoprotein-microRNA may be a promising disease biomarker. We investigated the stability of high-density lipoprotein-microRNAs in different storage conditions as this is an important issue for its application to the field of clinical research. Methods microRNAs were extracted from the high-density lipoprotein fraction that was purified from the serum. miR-135 a and miR-223, which are known to be present in high-density lipoprotein, were quantified by quantitative real-time PCR. The influence of preanalytical parameters on the analysis of high-density lipoprotein-miRNAs was examined by the effect of RNase, storage conditions, and freezing and thawing. Results The concentrations of microRNA in high-density lipoprotein were not altered by RNase A treatment (0–100 U/mL). No significant change in these microRNAs was observed after storing serum at room temperature or 4℃ for 0–24 h, and there was a similar result in the cryopreservation for up to two weeks. Also, high-density lipoprotein-microRNAs were stable for, at least, up to five freeze–thaw cycles. Conclusions These results demonstrated that high-density lipoprotein-microRNAs are relatively resistant to various storage conditions. This study provides new and important information on the stability of high-density lipoprotein-microRNAs.Keywords:
High-density lipoprotein
Low-density lipoprotein
The catabolism of very-low-density lipoprotein apoprotein B and its conversion to low-density lipoprotein was studied in five chow-fed cynomolgus monkeys following injection of radioiodinated homologous very-low-density lipoproteins. The mean (+/- SD) fractional catabolic rate of very-low-density lipoprotein apoprotein B was 0.97 +/- 0.20 h-1 and the mean (+/- SD) production rate was 0.76 +/- 0.20 mg X kg-1 X h-1. The percent of conversion of very-low-density lipoprotein apoprotein B to low-density lipoprotein ranged from 33 to 59%. In separate studies of low-density lipoprotein apoprotein B turnover performed using homologous radiolabeled low-density lipoprotein in five additional animals, the mean (+/- SD) fractional catabolic rate for low-density lipoprotein apoprotein B was 0.050 +/- 0.017 h-1 and the mean (+/- SD) apoprotein B production rate was 0.70 +/- 0.18 mg X kg-1 X h-1. Comparison of the total low-density lipoprotein apoprotein B production with that derived from very-low-density lipoprotein apoprotein B suggested that a large fraction of plasma low-density lipoprotein apoprotein B was derived from a source exclusive of circulating very-low-density lipoprotein apoprotein B. This was confirmed in two animals by simultaneous injection of radiolabeled very-low-density and low-density lipoproteins. Thus, a significant proportion of cynomolgus monkey low-density lipoproteins are produced either by direct hepatic secretion or by rapid conversion of lower-density lipoproteins before they appear in the peripheral circulation.
Catabolism
Low-density lipoprotein
High-density lipoprotein
Lipoprotein particle
Cite
Citations (39)
Low-density lipoprotein
High-density lipoprotein
Cite
Citations (20)
Cite
Citations (1)
Abstract Accumulation of rhodamine B isothyocyanate‐conjugated low density lipoproteins (R‐LDL) in cultured endothelial cells from human umbilical cord was studied with a fluorescence activated cell sorter. R‐LDL uptake was blocked at 0°C, inhibited by addition of excess of nonlabeled low density lipoprotein, high density lipoprotein‐2 or high density lipoprotein‐3. High density lipoprotein‐2 was about twice as effective in inhibition of R‐LDL uptake as high density lipoprotein‐3. Endothelial cells that formed a contact‐inhibited monolayer lost the ability to incorporate R‐LDL via a receptor‐mediated pathway. Using R‐LDL, it was possible to distinguish cells with different levels of R‐LDL incorporation.
Low-density lipoprotein
High-density lipoprotein
Cite
Citations (10)
Low-density lipoprotein
High-density lipoprotein
Intermediate-density lipoprotein
Cite
Citations (100)
Low-density lipoprotein
High-density lipoprotein
Lipoprotein particle
Cite
Citations (66)
Low-density lipoprotein
Aspartic acid
High-density lipoprotein
Intermediate-density lipoprotein
Glutamic acid
Cite
Citations (20)
4-Hydroxynonenal (HNE) is a major aldehydic propagation product formed during peroxidation of unsaturated fatty acids. The aldehyde was used to modify freshly prepared human low-density lipoprotein (LDL). A polyclonal antiserum was raised in the rabbit and absorbed with freshly prepared LDL. The antiserum did not react with human LDL, but reacted with CuCl2-oxidized LDL and in a dose-dependent manner with LDL, modified with 1, 2 and 3 mM-HNE, in the double-diffusion analysis. LDL treated with 4 mM of hexanal or hepta-2,4-dienal or 4-hydroxyhexenal or malonaldehyde (4 or 20 mM) did not react with the antiserum. However, LDL modified with 4 mM-4-hydroxyoctenal showed a very weak reaction. Lipoprotein (a) and very-low-density lipoprotein were revealed for the first time to undergo oxidative modification initiated by CuCl2. This was evidenced by the generation of lipid hydroperoxides and thiobarbituric acid-reactive substances, as well as by a marked increase in the electrophoretic mobility. After oxidation these two lipoproteins also reacted positively with the antiserum against HNE-modified LDL.
Low-density lipoprotein
Polyclonal antibodies
Thiobarbituric acid
Intermediate-density lipoprotein
4-Hydroxynonenal
Cite
Citations (80)
High-density lipoprotein
Reverse cholesterol transport
Low-density lipoprotein
Efflux
Apolipoprotein E
Cite
Citations (6)
The most accepted property of high-density lipoprotein is reverse cholesterol transport. However, other beneficial actions may contribute to the antiatherogenic role of high-density lipoprotein. This review addresses the action of high-density lipoprotein beyond reverse cholesterol transport.High-density lipoprotein cholesterol levels are inversely associated with coronary heart disease and other forms of vascular disease. Apart from transferring excess cholesterol to the liver, high-density lipoprotein exhibits favorable effects on oxidation, inflammation, thrombosis and endothelial function. Some of these actions are at least in part attributed to high-density lipoprotein-associated enzymes, such as paraoxonase and platelet-activating factor acetylhydrolase. However, high-density lipoprotein can become dysfunctional and proatherogenic under certain circumstances.Current data suggest that high-density lipoprotein possesses various properties beyond reverse cholesterol transport. However, many issues on the exact role of high-density lipoprotein remain unknown. Future research is needed.
High-density lipoprotein
Reverse cholesterol transport
Paraoxonase
Cite
Citations (200)