| Objective:1. To establish and evaluate a sandwich ELISA for detecting the concentration of apoM in human serum.And we measure the serum levels of apoM in healthy persons with this method and establish the reference ranges of apoM in healthy persons.2. We measure the serum levels of apoM in type 2 diabetes to explore if the serum apoM levels play a possible role for type 2 diabetes complicated atherosclerosis.Method:1.We have developed a sandwich ELISA with anti apoM monoclonal antibody for coated antibody, horseradish peroxidase (Horseradish Peroxidase, HRP) labeled anti apoM monoclonal antibody for enzyme-labeled antibody. This method is evaluated through a series of the methodology test.2. We have established reference values of apoM for healthy individuals in our laboratory through the measurement of 483 healthy samples.3.We measure the serum levels of apoM by enzyme linked immunosorbent assay(ELISA) in 117 patients with type 2 diabetes and 111 control subjects. The serum levels of hs-CRP by latex-enhanced immunoturbidimetric assay. And the lipid profiles are determined by Hitachi 7170A analyzer. Compared with the serum levels of apoM, hs-CRP and the lipid,then analyze the correlations between apoM and the other indicators in the two groups.Results:1.The linear range of this ELISA for detecting the concentration of apoM in human serum presents 31.25ng/ml～500ng/ml with the lowest detectable concentration 23.92ng/ml, the interference to apoM detection due to Lp(a), apoAI and apoB is small in our study. Between-assay variation and within-assay variation in the ELISA are less than 10%,respectively.2. The apoM reference interval for women is 22.55±11.60μg/mL,for men the reference range for apoM is 21.44±12.71μg/mL. The whole reference interval of apoM is 22.06±12.10μg/mL.In healthy individuals, the serum apoM levels are positively related to both TC(r=0.106,p<0.05) and HDL-C (r=0.395,p<0.01), but negatively correlated to TG(r=-0.186,p<0.05).3.The serum apoM concentrations are lower in type 2 diabetes compared to healthy controls(23.36±13.55μg/mL vs.26.86±10.40μg/mL),there is statistically significant difference between them (p <0.05).In type 2 diabetes, the serum apoM levels are positively related to both TC(r=0.235,P<0.05) and HDL-C (r=0.183,p<0.05). While the serum apoM levels are a negative correlation with the serum hs-CRP levels (r=-0.185,p<0.05). The correlations in control subjects are the same.Conclusion:1.We have established a sandwich ELISA with high sensitivity and specificity to measure the apoM concentration in human serum, and establish the reference range of apoM for healthy individuals.2.The serum levels of apoM are lower in type 2 diabetes compared to control group; in type 2 diabetes, the serum apoM levels are positively correlated with both TC and HDL-C, but negatively correlated with hs-CRP. These results suggest that apoM might be involved in the metabolism of cholesterol and HDL in vivo, moreover, apoM maybe participate in inflammation by inhibiting inflammatory factors such as hs-CRP, apoM maybe play a role in protecting against atherosclerosis. In conclusion,the lower serum apoM levels have a possible predictivity to type 2 diabetes complicated atherosclerosis. |
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