Orexin A hormone and its relation to Coronary heart diseases

 

Rana F. Jasim¹*, Thikra A. Allwsh²

¹Department of Chemistry, Collage of Education for Girls, University of Mosul, Mosul, Iraq.

²Department of Chemistry, Collage of Science, University of Mosul, Mosul, Iraq.

 

ABSTRACT:

The research included determination of the level of orexin-A hormone in coronary heart patients and compared to the control group. The results showed that the normal mean level of Orexin  A hormone in serum was (344. 47± 6.51pg/mol) in control group for both sexes with ages range between (20-more of 50) year and there was a significant decrease in level of Orexin A in coronary heart patients compared to control group. Also, it has been found that Orexin A level was not affected by age and sex, while there was a significant decrease in hormone level according to body mass index (BMI), also a significant decrease in hormone level in smoking patients compared to non-smoking has been found. The results showed a significantly increase of activity enzymes Peroxidase, Lactate dehydrogenase, Creatine Kinase  and concentration  of Cyststin-C, Copper, total cholesterol, triglyceride, low density lipoprotein-cholesterol and very low density lipoprotein-cholesterol also the results showed a significantly decreasing of Arylestase activity and concentration of high density lipoprotein-cholesterol in patients compared to control group. Correlation coefficients of Orexin A hormone with some clinical parameters were examined in control and patients. The results showed that Orexin A has a significant negative correlation with concentration of total cholesterol, triglyceride, LDL-C, VLDL-C, cystatin-c, copper and peroxidase activity in control and patients group, and with Iron in control group. Also, there was a significant positive correlation with concentration of HDL-C and Arylesterase activity in control and patients group. It was concluded that low level of Orexin A hormone is a risk of coronary heart disease and Orxine A has a critical role in coronary heart disease. Moreover, the level of the hormone could be used as a marker for coronary heart disease and its role of obesity.

 

 

 

INTRODUCTION:

Coronary heart disease (CHD) is one of the  major cardiovascular diseases affecting the world's population ¹. Although we are in the twentieth century, but coronary heart disease is still the leading cause of death and disease in all of the developed and developing countries². There may be a correlation between coronary heart disease and the detected hormone Orexin A³.

 

Orexin A protect against atherosclerosis by inhibition the releasing of colony stimulating factor 1(CSF1) from neutrophils cells, CSF1 increase macrophage cells which have a pro-inflammatory functions^4,5.

 

Orexin, also known as hypocretins, was first discovered in the hypothalamus in 1998 by two groups of researchers. They found the same new peptides using different methods⁶, One group called  it Orexin, came  from the Greek word "Orexis" which means appetite, and the other group called it  "Hypocretin" since it showed  sequence of  amino acid similar to secretin, the gut hormone⁷

 

Orexin is a neuropeptide generated by neurons distributed in lateral hypothalamic area. There are two forms of Orexins, Orexin-A (OXA) (consist of 33 amino acid with two intra-chain disulfide bond, molecular weight 3562 Da) and Orexin B(OXB) (liner peptide contain 28 amino acid which are 46% identical with OXA, molecular weight 2937 Da)⁸. Researchers found that OXA is of great importance biologically than OXB⁶.

 

 

Orexin neurons spreads significantly throughout the brain and the spinal cord⁹. Orexin have several physiological functions, Orexin  is known to play an important  role in controlling  appetite and increasing mammalian food intake¹⁰, Also OXA regulate the secretion of endocrine, metabolism and the function of the heart and blood vessels³.   

 

AIM OF RESEARCH:

Since there are few previous studies in Iraq about OXA hormone, so it was suggested to study it in control group and coronary heart patients and the relationship between OXA and some clinical parameters.

 

MATERIAL AND METHODS:

This  study included (100) healthy subjects (49 Females, 51 Males),with age between (20-50< year) and (125) patients (64 Females,61 Male) with coronary heart disease from Ibn- sena education hospital- cardio care unit (CCU) and  AL-Salam education hospital-intensive care unit (ICU) in Mosul city at age between (36-50<year)  with coronary heart disease. Fasting and non-fasting blood were taken, and the serum were separated and used to estimate the following clinical parameters:

1.   Orexin-A hormone: Was measured by competitive-enzyme linked immunosorbent assay (ELISA) technique¹¹ using Elabscience biotechnology, INC Kit (USA), and the analysis was performed in the immunity laboratory in Al-Salam hospital in Mosul city using  BIO-TEK INSTRUMENT, INC, USA.

2.   Cystatin-C concentration: Was determined by immunochromatographic rapid method using Veda lab kit. This analysis was performed using VEDALAB Easy Reader+^®, CE, France.

3.   Lactate dehydrogenase (LDH) activity: Was determined by SFBC modified method using BIOLABO Kit (France).

4.   Creatine Kinase (C.K) activity: Was determined by enzymatic colorimetric method using RANDOX kit (U.K).

5.   Arylesterase activity: Was determined by enzymatic hydrolysis of phenyl acetate to phenol and acetic acid^12,13.

6.   Peroxidase activity: Was determined by enzymatic oxidation of hydrogen peroxidase¹⁴.

7.   Copper and Iron concentration: Were determined by atomic adsorption technique using atomic adsorption spectrometer, A Analyst 400, in administrative institute-Department of pathological analyzes.

8.   Total Cholesterol (TC): Was determined by enzymatic colorimetric method using BIOLABO Kit (France).

9.   Triglycerides (TG): Was estimated by enzymatic colorimetric method using BIOLABO Kit (France).

 

10. High density lipoprotein-cholesterol (HDL-C): Was estimated by precipitation method using BIOLOABO kit (France).

11. Low density lipoprotein-cholesterol (LDL-C): Was calculated by following friedewald equation: LDL-C (mmol/L) =Total cholesterol–HDL-C –TG/2.2¹⁵.

12. Vary low density lipoprotein –cholesterol (VLDL-C): Was calculated using the equation: VLDL-C(mmol/L) =TG (mmol/L)/2.2¹⁶.

13. Calculation of atherogenic index (AI): Was calculated using the equation: AI =log (TG/HDL-C)¹⁷.

14. Calculation of antiatherogenic index (AAI): Was calculated using the equation: AAI =(HDL×100/TC-HDL-C)¹⁸.

15. Body Mass Index (BMI): Was calculated as weight in kilogram divided by square height in meters¹⁹.

 

DATA ANALYSIS:

The data obtained in this study were analyzed using Statistical Package for Social Sciences (SPSS)

·       Standard statistical methods were used to determine the mean and standard error.

·       T-test was used to compare between two parameters.

·       One-way Anova is used to compare between more than two parameters.

·       Liner regression analysis [Pearson correlation coefficient (r)] was used to find the        

·       Relationship between different chemical parameters.

·       P-Value ≤ 0.05 was considered to be statistically significant²⁰.

 

RESULT AND DISCUSION:

1-Orexin Level in coronary heart patients compared to control group:

The results in table (1) showed that the normal mean level of OXA hormone is (344.47±6.51pg/ml) in control group. This was approximate to the results showed by²¹ who showed that the normal levels of OXA in normal subjects was (338.48±20.24pg/ml) and consistent with the range (230-376 pg/ml) found by²². The results also showed that the coronary heart patients have a significantly lower Orexin A level compared to control group as in figure (1).

 

Figure (1): Orexin A level at coronary heart patients compared to control

These results were in agreement with^4,5. The cause of these decreases in coronary heart patients might due to increase in lipids in atherosclerosis²³, or might be also due to increase of oxidative stress which cause reduce in Orexin A level by effecting on neurons containing Orexin¹⁰ or due to stress which cause inhibition of Orexin²⁴.

 

Table (1): The Level of Orexin A hormone in control group and coronary heart patients according to age and sex

**Significant difference at p≤ 0.01, S.E.= Standard error.

 

2-Orexin A Level in coronary heart patients compared to control group according to the age and sex:

The results in table (1) showed that there were a gradual decrease non-significantly in Orexin A Level with age for each sexes in coronary heart patients and control group, these results were coordinate with those found by¹¹.

 

3-Orexin A Level in coronary heart patients compared to control group according to BMI:

In table (2) the results showed that there was a significant decrease in Orexin A level in over weight and obese compared with under and normal weight in control and patients group. These results were in consistent with²⁵ that Orexin A level are negatively correlated with BMI in human. Orexin A promote obesity resistance through enhance spontaneous physical activity and energy expenditure⁸ so mice with deficient Orexin neuron have obesity of  late-onset  despite reduced food intake, also Orexin enhancing energy expenditure through modulating locomotor activity and brown adipose tissue thermogenesis²⁶.

 

Table (2): The Level of Orexin A hormone in control group and coronary heart patients according to BMI

* Significant difference at p≤ 0.05   ** Significant difference at p≤ 0.01   

 

4-Orexin A Level in coronary heart patients compared to control group according to the smoking:

The results in table (3) showed that there was a significant decrease in Orexin A level in smoker compared to non-smoker  and no significant differences in quit smoker compared to non-smoker in patients and control group, these results were in agreement with those found by²⁷ the cause of reduction due to the effect of nicotine on Orexin function by influence on weight regulation²⁸ also due to the negative correlation between nicotine craving and Orexin levels²⁷.

 

Table (3): The Level of Orexin A hormone in coronary heart patients according to smoking

* Significant difference at p≤ 0.05

 

The concentration of some clinical parameters in control group and coronary heart patients:

The research also involves an estimation of some clinical parameters in coronary heart patients compared with control group to find the relationship between Orexin A and some clinical parameters:

 

1-Cystatin C concentration:

Cystatin C is an inhibitor of cysteine protease produced by all cells and secreted in to the blood stream, filtered through the glomerulus. The results in table (4) demonstrate that the patients have a significant increase in cystatin C compared to control. These results were in agreement with^29,30, the cause due to an imbalance between Cystatine protease and Cystatin C in atherosclerotic lesion occur during atherogenesis²⁹  also due to association of Cystatin C with inflammation and atherosclerosis that Inflammatory cytokinase and atherosclerosis stimulate the production of  lysosomal cathepsin that contribute to disruption of plague, and Cystatin C is inhibitor of cathepsin, so increase Cystatine C concentration may be associated with inhibition of these cathepsins³¹.

 

2-Creatine Kinase (C.K) activity:

The results in table (4) showed a significantly increasing in C.K activity in patients compared to control group, these results were consistent with those found by³². The cause due to myocardial necrosis and death of myocardium cell which cause efflux addition amounts of enzyme³³.

 

3-Lactat Dehydrogenase (LDH) activity:

LDH is one of enzymatic biomarkers in the diagnosis of CHD in patients who reach hospital³³, the results in table (4) show a significant increase in LDH activity in patients compared to control group. These results were in agreement with those found by³⁴ the cause of LDH elevation is myocardial necrosis and release of enzyme from myocardial itself and then appears in serum.

 

4- Peroxidase activity:

As showed in table (4) a significantly increasing in serum peroxides activity in the patients compared to control group, this might be due to the increase of oxidative stress in coronary heart patients and increase reactive oxygen species³⁵.

 

5- Arylesterase activity:

There was a significant decrease in arylesterase activity in the patients compared to control group as shown in table (4). These results were in agreement with those found by^13,35,36 the cause might be due to reduction in HDL level since arylesterase associated to HDL which decreased in patients.

 

6- Copper and Iron concentration:

The concentration of copper (Cu) showed a significant increase in patients compared to control group as shown in table (4), these results were consistent with  those found by³⁷ the cause may be due to the pro-oxidation effect of Cu by fenton-type redox reaction, which result in oxidative cell damage³⁷. While there was no significant difference in Iron concentration between patients and control group.

 

7-Total Cholesterol (TC), Triglyceride (TG), VLVL-C, LDL-C and HDL-C concentration:

The results in table (4) showed a significant increase in TC, TG, VLDL-C and LDL-C concentration while there was a significant decrease in HDL-C concentration in patients compared to control group, these results were in agreement with^13,35,38.

 

8- Atherogenic (AI) and antiatherogenic (AAI) index:

Also there was a significant increase in atherogenic index in patients compared to control group and these results due to increase TG and decrease HDL-C in patients compared to control group¹⁷. The results also demonstrate a significant decrease in antiatherogenic index in patients compared to control group because of decrease HDL-C and increase TC in patients compared to control group^35,38.

 

Table (4): The concentration of some clinical parameters in control group and coronary heart patients

* Significant difference at p≤ 0.05   ** Significant difference at p≤ 0.01  ***Significant difference at p≤ 0.001      

Correlation between level of Orexin A hormone and some parameters in control group and coronary heart patients:

As showed in table (5) there was a negative correlation non-significantly between Orexin A level and C.K and LDH activity. While a negative correlation significantly  has been found between cystatin C concentration in control group and patients, The cause might by due to that the Orexin A has an anti-inflammatory  effect and ameliorate inflammatory markers³⁹ and there is close correlation between Cystatin C and inflammatory marker or it may be due to that Cystatin C is a sensitive marker of renal dysfunction in early state which may be involved in CHD process and Orexin A has a renoprotective effect³⁰.

 

On the other hand the results in table (5) showed that  there was a significant  negative correlation between Orexin A level and Peroxidase activity in control and patients group, the cause might be due to elevated oxidative stress which cause increase in reactive oxygen species such as H₂O₂  and increase in Peroxidase activity⁴⁰ while oxidative stress cause decrease in Orexin A level¹⁰. Also it has been found a appositive correlation between Orexin A level and arylesterase activity in control and patients group, this can be attributed to the oxidative stress which decrease the activity of enzyme and Orexin A level^13,35. The results in table (5) showed a significant negative correlation between Orexin A level and the concentration of Copper in control and patients group and the concentration of Iron in control group, this might be due to the oxidative effect of these mental which cause decrease in Orexin A level³⁷.

 

Table (5): The correlation between Orexin A hormone level, and Clinical parameters in control group and coronary heart patients  

*Correlation is significant at p ≤ 0.05 

**Correlation is significant at p ≤ 0.01

 

A significant negative correlation appeared between orexin A level and the concentration of TC, TG, VLDL-C and LDL-C  in control and patients group, these results in agreement with that found by⁴¹, this might be due to the possible direct role for Orexin in adipose tissue metabolism, Orexin A  decrease hormone sensitive lipase (HSL) in adipose tissue and inhibit lipolysis also decrease glycerol release from adipose tissue by re-esterification of fatty acid⁴² also Orexin A increase insulin-stimulated glucose uptake and increase lipogenesis by enhance the rate of glucose uptake⁴³. On the other hand, a significant positive correlation was demonstrated between Orexin A level and the concentration of HDL-C in control and patients group, these result is consistent with⁴⁴ and this might be due to the effect of Orexin A which cause increase lipoprotein lipase activity⁴³ or may be due to that Orexin A promote adiponectin which correlation positively with HDL-C⁴⁵.

 

Finally, it has been demonstrated a significant negative correlation between Orexin A level and AI in control and patients group, these results due to negative correlation between Orexin A level and the concentration of TG and positive correlation between Orexin A level and the concentration of HL-C^41,44. While there was a significant positive correlation between Orexin A level and AAI in control and patients group, these results due to positive correlation between Orexin A and HDL-C and negative correlation between Orexin A and TC^41,44.

 

CONCLUSIONS:

It was concluded that Orexin A hormone has a major role in coronary heart disease and there is relationship between Orexin A and BMI and could be used as a marker for coronary heart disease and obesity.

 

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Accepted on 16.06.2020           © RJPT All right reserved

Research J. Pharm. and Tech 2021; 14(3):1417-1422.