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Investigation of plasma amyloid beta 1-42 levels and its correlation with some cardiovascular risk factors in people over forty years old

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Objectives: Amyloid beta 1-42 (Aβ 1-42) is related to several disorders including nervous, cardiovascular and metabolic disorder. We investigated plasma Aβ 1-42 level and examined its correlation with some cardiovascular risk factors.

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INVESTIGATION OF PLASMA AMYLOID BETA 1-42 LEVELS

AND ITS CORRELATION WITH SOME CARDIOVASCULAR RISK FACTORS IN PEOPLE OVER FORTY YEARS OLD

Nguyen Minh Hai*; Nguyen Minh Nui*; Ho Anh Son**

SUMMARY

Objectives: Amyloid beta 1-42 (Aβ 1-42) is related to several disorders including nervous, cardiovascular and metabolic disorder We investigated plasma Aβ 1-42 level and examined its correlation with some cardiovascular risk factors Subjects and methods: Level of Aβ 1-42 was measured in plasma of 40 people over forty years old without type 2 diabetes mellitus and hypertension by ELISA method Results and conclusions: Average plasma Aβ 1-42 level was 98.88 ± 92.45 pg/mL, median was 65.49 pg/mL with wide range from 45.83 to 494.32 pg/mL There was no significant correlation between plasma Aβ 1-42 level and cardiovascular risk factors such as age, plasma lipids

* Key words: Metabolic disorder; Amyloid beta

INTRODUCTION

Amyloid is a specific peptide family

including aggregating beta chains Small,

focal, clinically silent amyloid deposits in

the brain, heart, seminal vesicles, and joints

is a universal accompaniment of aging

These results in amyloidosis, structural

disorders, reduction and loss in function of

various organs in the body The underlying

molecular abnormalities may be either

acquired or hereditary and about 20 different

proteins can form clinically or pathologically

significant amyloid fibrils in vivo Brain

amyloid deposit is a major pathological

feature of Alzheimer disease Islet amyloid

deposits trigger type 2 diabetes mellitus

Amyloid also deposits in joints, peripheral

arteries, kidneys Aβ 1-42 is one of amyloid

molecular, which is considered to relate

with different diseases such as Alzheimer’s

disease, diabetes, cardiovascular diseases [2] Although many studies about Aβ 1-42 have been performed, its role and physical function is still unknown So far in Vietnam, there has been no investigation about the exact physical function of Aβ 1-42 Therefore, we carry out this study to:

- Examine the level of Aβ 1-42 plasma

in the people over 40 years old

- Investigate whether there is any relationship between plasma Aβ 1-42 level and cardiovascular risk factors

SUBJECTS AND METHODS

1 Subjects

* Selection criteria:

Study was carried out on 40 people over forty years old without type 2 diabetes

mellitus and hypertension

* 103 Hospital

** Military Medical University

Corresponding author: Nguyen Minh Nui (minhnuinguyen@gmail.com)

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2 Methods

- Cross-sectional study without control

group

- All people were clinically and paraclinically

examined by measuring weight, height,

blood pressure, level of glucose, creatinine,

acid uric, lipid in their blood Level of

Aβ 1-42 plasma was measured by ELISA

method in Physiopathology Department,

Vietnam Millitary Medical University

- Methods of Aβ 1-42 measurement [9]:

+ 5 mL of whole bood was taken into

tubes with anticoagulant (citrate) in the

morning Afterward, the sample was

centrifugated for 20 min at 2,000 rpm

Collect the supernatant carefully as plasma

sample If precipitate was seen during

reservation, the sample should be centrifugated

again

+ The Microelisa stripplate provided in

the ELISA kit has been pre-coated with an

antibody specific to Aβ 1-42 Standards or

samples were added to the appropriate

Microelisa stripplate wells and combined

to the specific antibody Then a Horseradish

Peroxidase (HRP) - conjugated antibody

specific for Aβ 1-42 was added to each

Microelisa stripplate well and incubated

Free components were washed away before

adding the Chromogen solution A and

Chromogen solution B to each well Blue

only appreared in those wells that contain

Aβ 1-42 and Horseradish peroxidase

conjugated Aβ 1-42 antibody and then it

turned yellow after addition of the stop

solution The optical density is measured

spectrophotometrically at a wavelength

of 450 nm The optical density value is

proportional to the concentration of Aβ 1-42 The concentration of Aβ 1-42 in the samples is calculated by comparing the optical density of the samples to the standard curve

* Statistical analysis: to initially explore the relationship between variable sets, scattergram blots using Microsoft Excell (2010), where a relationship was found Stata verson 12.0 was used to verify the statistical significances of the association Simple linear regression was initially used Pearson chi-square was used to determine the association between concentration

of Aβ 1-42 plasma and baseline participant characteristics

RESULTS

1 Characteristics of the subjects

40 people included 24 males (60%) and

16 females (40%) Mean age was 54.38 ±

11.23 years

2 Plasma Aβ 1-42 levels

Table 1: Plasma Aβ 1-42 level

Variable

Level of Aβ 1-42 plasma

(pg/ml)

Mean ± SD Median

(Min - max) Male (n = 24) 83.77 ± 58.17 64.46

(45.83 - 313.64) Female (n = 16) 121.55 ± 126.97 66.67

(47.59 - 494.32) Summarization 98.88 ± 92.45 65.49

(45.83 - 494.32)

Both mean and medium plasma Aβ 1-42

in male were not significantly lower than those in female (p > 0.05) Levels of Aβ 1-42 fluctuated from 45.83 pg/mL to 494.32 pg/mL

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3 Association between plasma Aβ 1-42 levels with age, blood glucose and

acid uric levels

* Association between plasma Aβ 1-42 levels and age:

Figure 1: Correlation between plasma Aβ 1-42 levels and age

No significant correlation between plasma Aβ 1-42 levels and age of participants was

found (p > 0.05)

* Association of plasma Aβ 1-42 level with blood glucose and acid uric levels:

A = 5.6765 * Glucose + 71.579

r = 0.043, p= 0.79

0

100

200

300

400

500

600

Glucose (mmol/L)

A = -0.311 * Uric + 209.48

r = 0.251, p = 0.11

0 100 200 300 400 500 600

Acid uric (µmol/L)

A B

Figure 2: Correlation between plasma Aβ 1-42 levels and glucose (A),

acid uric (B) levels

Plasma Aβ 1-42 levels did not correlate with either glucose or acid uric levels (p > 0.05)

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4 Association between plasma Aβ 1-42 levels with total cholesterol, HDL-C, LDL-C, and triglyceride levels

A = -20.324 * Cholesterol + 208.69

r = -0.022, p = 0.61

0

100

200

300

400

500

600

Cholesterol (mmol/L)

A = -7.332 * Triglyceride +114.19

r = -0.138, p = 0.43

0 100 200 300 400 500 600

Triglyceride (mmol/L)

A B

A= 111.17 * HDL - 40.721

r = 0.327, p = 0.08

0

100

200

300

400

500

600

HDL-C (mmol/L)

A = -37.177 * LDL + 222.65

r =-0.34, p = 0.44

0 100 200 300 400 500 600

LDL-C (mmol/L)

C D

Figure 1: Correlation between plasma Aβ 1-42 levels, cholesterol (A), triglyceride (B),

HDL-C (C) and LDL-C (D)

No significant correlation of plasma Aβ 1-42 plasma levels with cholesterol, triglyceride, HDL-C and LDL-C was observed

DISCUSSION

Aβ 1-42 is supposed to relate to a variety

of disorders such as Alzheimer’s disease,

type 2 diabetes mellitus In the body, Aβ 1-42

can be solute in cerebrospinal fluid and blood or deposits in brain, pancreas

We investigated whether there were any associations between plasma Aβ 1-42 levels and cardiovascular risk factors

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In our study, the mean (±SD) and median

plasma Aβ 1-42 were 98.88 ± 92.45 (pg/mL)

and 65.49 (pg/mL) The concentration of

Aβ 1-42 plasma fluctuated in a large range

between 45.83 (pg/mL) and approximately

500 (pg/mL) It was interesting to note that

participants who had the highest plasma

Aβ 1-42 level had normal levels of lipid,

glucose, acid uric in the blood with total

cholesterol = 5.17 (mmol/L), HDL-C = 1.8

mmol/L, LDL-C = 3 (mmol/L), LDL-C = 3

(mmol/L), triglyceride = 0.54 (mmol/L), acid

uric = 327 (µmol/L)

In the study by Kelvin et al, mean (±SD)

and medium of plasma Aβ 1-42 levels in

a group of 18 healthy adults were 29.1 ±

18.1 (pg/mL) and 27 (pg/mL) These results

were lower than our findings and the

concentration of plasma Aβ 1-42 levels

fluctuated in narrower range between 2.5

(pg/mL) and 62 (pg/mL) However, mean

age of the participants in Kelvin’s research

was 36, which was lower than the figure

in our study [4]

In 2000, Mehta and his colleagues

conducted a study on the patients older

than forty with Alzheimer’s disease and

control subjects Levels of Aβ 1-42 plasma

were measured by a sandwich

enzyme-linked immunosorbent assay Both the

medium of plasma Aβ 1-42 levels in group

of Alzheimer’s disease and control group

were higher than the results of our study

with 73 (pg/mL) (minimum = 25 pg/mL,

maximum = 880 pg/mL) and 81 (pg/mL)

(minimum = 25 pg/mL, maximum = 905

pg/mL), respectively The plasma Aβ 1-42

levels also fluctuated in a large range [6]

Metti and his colleagues (2012) used Innogenetics INNO-BIA assays to measure plasma Aβ 1-42 level in 988 community-dwelling elders who suffer from diabetes, stroke, myocardial infarction and hypertension Mean age of these patients was 74.0 ±

3 years The mean (±SD) and median of plasma Aβ 1-42 levels were 33.9 ± 9.7 and 32.83 (pg/mL) [7] The mean and plasma

Aβ 1-42 levels in our study were both similar

to Mehta and Metti’s findings Thus, the differences in plasma Aβ 1-42 levels in these studies were affected significantly

by age

High plasma cholesterol, triglyceride and LDL levels, as well as low plasma HDL levels have long been known to be associated with cardiovascular risk factors and have been identified as significant risk factors for Alzheimer’s disease HDL has been shown to carry Aβ 1-42 in plasma and cerebrospinal fluid [1] Plasma HDL has been reported to be lower in patients with Alzheimer’s disease and is highly correlated with lower cognitive scores from these patients [1, 3] Many studies indicated that HDL which interact with

Aβ 1-42 in plasma, can regulate Aβ 1-42 aggregation, degradation and toxicity [5, 8] Our study did not show significant associations between levels of total cholesterol, triglyceride, LDL-C or HDL-C and Aβ 1-42 levels This results were similar to those of Kelvin et al in 2005 [4] However, our study was performed in healthy individuals Whether there is any correlation between Aβ 1-42 levels and cardiovascular risk factors in diabetic patients

as well as in healthy people, which need

to be investigated in the future

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CONCLUSION

The mean and medium of plasma Aβ 1-42

levels in people over forty years old were

98.88 ± 92.45 (pg/mL) and 65.49 (pg/mL)

and plasma Aβ 1-42 levels fluctuated in a

large range between 45.83 (pg/mL) and

494.32 (pg/mL) No correlations between

Aβ 1-42 levels and several cardiovascular

risk factors such age, plasma glucose,

acid uric and lipids levels were observed

REFERENCES

1 Cole G M, Beech W, Frautschy S A et al.

Lipoprotein effects on Abeta accumulation and

degradation by microglia in vitro J Neurosci Res

1999, 57 (4), pp 504-520

2 David A Warrell, Timothy M Cox, Firth

Oxford University Press 2003, pp.163-167

density lipoprotein decreases beta-amyloid

toxicity in cortical cell culture Neuroreport

1997, 8 (5), pp.1127-1130

4 Kelvin Balakrishnana, Giuseppe Verdilea,

positively with increased body fat in healthy individuals Journal of Alzheimer’s Disease

2005, 8, pp.269-282

5 Kuo YM, Emmerling MR, Bisgaier CL et al

Elevated low-density lipoprotein in Alzheimer's disease correlates with brain abeta 1-42 levels Biochem Biophys Res Commun 1998, 252 (3), pp.711-715

6 Mehta PD, Pirttila T, Mehta SP et al.

Plasma and cerebrospinal fluid levels of amyloid beta proteins 1-40 and 1-42 in Alzheimer disease Arch Neurol 2000, 57 (1),

pp 5-100

7 Metti AL, Cauley JA, Newman AB et al.

Plasma beta amyloid level and depression in older adults J Gerontol A Biol Sci Med Sci

2013, 68 (1), pp.74-79

lipoprotein inhibits assembly of amyloid beta-peptides into fibrils Biochem Biophys Res Commun 2000, 270 (1), pp.62-66

9 Schmidt S, Matthew J, Nixon R, Mathews

immunosorbent assay in Sigurdsson, E.M., Calero, M & Gasset, M (eds.) Amyloid Proteins Methods in Molecular Biology: Humana Press 2012, pp.507-527

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