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Latin American Journal of Biotechnology and Life Sciences
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2021.06.02.9
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INVESTIGATION / RESEARCH
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The effect of consuming Pokea clam meat on nitric oxide plasma levels in hypertensive patients in Sampara District, Konawe District


 
 
I Putu Sudayasa1, Suryani As’ad1, Rosdiana Natsir1, Venny Hadju1,  Mochammad Hatta1, Muh. Nasrum Massi1, Burhanuddin Bahar1, Sri Rahmadhani1, Yusminah Hala2,La Ode Alifariki3*
Available from: http://dx.doi.org/10.21931/RB/2020.06.02.9
(registering DOI)

 
 
ABSTRACT

 
The high number of vitamins and minerals in Pokea meat encouraged us to determine the effect of consuming Pokea meat on Nitric Oxide Plasma levels in patients with hypertension and normotension. This study aimed to analyze Pokea meat consumption (Batissa violacea var. celebensis von Martens) on plasma oxide (NO) levels in hypertensive patients in Sampara, Konawe District. This research uses an observational analytic method with a case-control study design through molecular biology approach. The sample comprises 60 people consisting of 30 case samples and 30 control samples using the purposive sampling technique. Laboratory examination data is on NO plasma levels. Statistical analysis used data analysis use-dependent t-test. The distribution of Pokea meat consumption variables in the Hypertension group respondents had a mean value of 35.14 ± 17.66, while in the Non-Hypertension group of respondents was 41.10 ± 19.82. In the variable nitric oxide, the Hypertension group had a mean and standard deviation of 69.48 ± 42.78 µmol / L while the Non-Hypertension group had a mean and standard deviation of 262.8 ± 39.90 µmol / L. The statistical test analysis results showed an effect of Pokea consumption on plasma NO levels (p = 0,000). Pokea Consumption Influences NO Plasma Levels in Hypertension Patients, and there are also differences in NO Plasma Levels in Patients with Hypertension and non-hypertension in Sampara District, Konawe District, Southeast Sulawesi.
Key-words: hypertension, nitric oxide plasma, Pokea clam, sampara
 
 
 
INTRODUCTION

 
 
Pokea clams or Batissa violacea var. celebensis von Martens (1897) are bivalves belonging to the Corbiculidae family and are a type of freshwater clam fish located in the Southeast Sulawesi region 1. Pokea is scattered in several large islands in Indonesia such as (West Papua, Sumatra, Sulawesi, Java), while in Sulawesi, particularly in Southeast Sulawesi's waters, especially in large rivers such as Pohara River, Lasolo River, Roraya River, Laeya River 2.
 
In 100-gram clam meat contains Clam meat is rich in omega-3 and omega-6 fatty acids and contains 59 kilocalories of energy 3. The amount of protein in shellfish is an average of 8 grams. This protein plays an essential role in the formation of enzymes in the body, the formation of organ cells and muscles, the formation of hormones, repair damaged cells in the body, regulate metabolism, form the immune system, and as a source of energy 3,4.
 
Yenni and colleagues' research regarding the proximate analysis of Pokea clam meat showed 50.48% protein content, 6.86% fat, 29.13% carbohydrate, and 5.53% fiber, and 2.70% water. Pokea meat contains the highest protein content of more than 50%, the moderate fat content of more than 5%, and more than 20% of carbohydrate content based on its dry weight. The nutritional content of Pokea is equivalent to several types of mollusks, which are empirically believed to be aphrodisiacs, increase reproductive vitality, treat fever, jaundice, and reduce blood pressure overcome hypertension 5.
 
Suryana, in her study, found a significant difference in respondents' Nitric Oxide levels in the Non-Hypertension and Hypertension groups with a value of p <α (p = 0.023). The mean value of Nitric Oxide levels in the Hypertension group was lower than Non-Hypertension. Likewise, Tadei and colleagues' results also show that the mechanism of blood vessel endothelial dysfunction is associated with a decrease in plasma NO levels 6.
 
Hypertension is ranked first among the 10 highest non-communicable diseases compared to acute respiratory infections in Southeast Sulawesi. Specifically, in the City of Kendari, hypertension ranks the third most disease 7.
 
The high number of vitamins and minerals in Pokea meat encouraged us to determine the effect of consuming Pokea meat on Nitric Oxide Plasma levels in patients with hypertension and normotension.

 
 
METHODS

 
 
This study is an observational analytic with a case-control design conducted in March-August 2019, in the Sampara District area. Using a purposive sampling technique, 60 respondents were selected, consisting of 30 case samples and 30 control samples. Sources of data regarding hypertension patients were obtained from the Sampara Health Center. Blood pressure confirmation is measured with a Sphygmomanometer. The research instrument was a Pokea meat consumption questionnaire, Semi-Quantitative Food Frequency Questionnaire (FFQ-SQ). Nitric Oxide Plasma levels are measured through venous blood collection and examined at the Hasanuddin University clinical laboratory approved by the Medical Faculty of Hasanuddin University (230/UN4.6.4.5.31/PP36/2019). Data analysis used a dependent t-test. Based on the Kolmogorov Smirnov test using SPSS V. 16.00, the data were normally distributed.
 


 
 
RESULTS

 
 
Analysis of general characteristics of respondents
 
Distribution of respondent characteristics based on gender, age group, level of education, occupation, ethnicity, residence, as listed in Table 1.
 
 

 
Table 1: Frequency Distribution of Respondent Characteristics

 
 
Table 1 shows the frequency distribution based on respondents' characteristics that the most gender in the Hypertension group was 19 women (40.4%), and in the Non-Hypertension group, the most female sex was 28 people (50.6%).
 
The most dominant age group in the Hypertension group is 40-46 years, as many as 8 people, and in the Non-Hypertension group, most at 40-46 years, as many as 10 people. Most types of work are housewives with 15 people and 26 people in the Non-Hypertension group. In the education category, the highest number of respondents in the Hypertension group was SMA, as many as 15 people in the Non-Hypertension group as many as 13 people are in SMA level of education.
 
Based on ethnicity, the most respondents were Tolaki tribe, with 27 people in the Hypertension group and 29 people in the Non-Hypertension group. Respondents in the Hypertension group mostly lived in Andepali and Andadowi villages with 10 people, while respondents in the Non-Hypertension group mostly lived in Andepali Village with 19 people. The respondents' general characteristics will have the distribution of particular characteristics from the examination of lipid profiles, plasma NO levels, and eNOS gene expression
 
.
 
Analysis of Special Characteristics of Respondents
 
Specific characteristics regarding the results of examining Pokea meat consumption, plasma NO levels, as listed in Table 2

 
 

 
Table 2: Distribution of Pokea meat consumption pattern and plasma Nitric Oxide levels in Hypertension and Non-Hypertension

 
 
Table 2 shows the distribution of Pokea meat consumption variables where respondents in the Hypertension group had a mean value of 35.14 ± 17.66, while in the Non-Hypertension group, it was 41.10 ± 19.82. The results also showed the amount of Pokea meat consumption in the Hypertension group was 13.25 gr at the minimum number and 67.86 at the maximum, while in the Non-Hypertension group, it was 13.25 for the minimum and 78.57 for the maximum.
 
Nitric oxide levels in the Hypertension group had Mean and Standard Deviations of 69.48 ± 42.78 µmol / L while those in the Non-Hypertension group had mean and standard deviations of 262.8 ± 39.90 µmol / L. The minimum and maximum values ​​of nitric oxide levels in the Hypertension group were 16.16 ± 255.46 µmol / L while in the Non-Hypertension group with values ​​87.83 ± 309.55 µmol / L.
 
 
 
Table 3: Analysis of the Effects of Pokea meat consumption on plasma Nitric Oxide levels in hypertensive patients
 
 
Table 3 shows the effect of Pokea meat consumption on nitric oxide levels with a mean value of 69,481 and p of 0,000.
 


 
Figure 1: Differences in levels of Nitric Oxide Plasma in patients with hypertension and not hypertension
 
 
Figure 1 shows the differences in nitric oxide levels (NO) in the Hypertension and Non-Hypertension groups, where the levels of nitric oxide (NO) in the Hypertension group are lower than those in the hypertension respondents.

 
 
DISCUSSION

 
 
Distribution of consumption of Pokea meat, nitric oxide plasma levels
During the initial interview with respondents (Hypertension and Not-Hypertension), it was revealed that 66.7% consumed Pokea meat 1-3 times and> 3 times a day as much as 33.3% of respondents. We also found that the Non-Hypertension group had a higher Pokea meat consumption pattern of 41.10 grams per day than the Hypertension group with a consumption pattern of 35.14 grams per day.
 
Based on the results of our observations on the community in the village of Andepali, Sampara District, Konawe District, 85% of respondents found that they often consumed Pokea meat in the last three months. with a proportion of 58% of respondents stating that they eat Pokea clam meat as much as 1-3 a day and as many as 57% consume Pokea with a portion of ½ -1 cup. Around 46% of the community obtains Pokea from the market or traveling vendors, and the others 78% process Pokea directly. In general, the type of processed Pokea consumed is in the form of satay Pokea (39%) and boiled Pokea (8%). The community manages Pokea meat by boiling 67%, processing it by frying 43%, sautéing 33% 8.
 
Research conducted by Rasyid, et al. on the chemical composition of Pokea meat in the Sampara area, Konawe Regency, Southeast Sulawesi obtained results from t-test analysis of the essential amino acids content of fresh and boiled Pokea meat; the only lysine was significantly different. The study also found non-essential amino acids, glutamic acid, serine, glycine, and cysteine, which differed significantly in composition in Pokea meat 9.
 
The total amount of amino acids in Pokea meat is relatively the same after boiling, while non-essential amino acids decreased significantly by 29.43%. In fresh meat and boiled Pokea identified as many as 25 types of fatty acids. The content of fatty acids in Pokea skin after boiling is relatively stable. Based on n6 / n3 and PUFA / SFA ratio, the ratio of fresh, boiled, and dry Pokea peels can be said to be healthy and safe for consumption. Identifying the extract is correct because it is proven to provide positive reaction results containing protein and carbohydrates 9.
 
Consumption of foods high in saturated fatty acids increases cholesterol levels in the blood and leads to atherosclerosis, affect the decrease in Nitric Oxide levels, leading to hypertension. Initially, LDL will be oxidized by superoxide produced by NAD (P) H macrophages oxidize. Ox-LDL stimulates several redox-sensitive processes that harm endothelial function. Through inhibition of eNOS activity and NO inactivation, ox-LDL may reduce NO bioavailability 10.
 
There was a difference between hypertension and non-hypertension groups in the variable level of oxide, where the non-hypertensive group was significantly higher than the hypertension group. This study's results are supported by previous research, which shows significant differences in respondent's Nitric Oxide levels in the Non-Hypertension and Hypertension groups with a value of p <α (p = 0.023). The mean value of Nitric Oxide levels in the Hypertension group was lower than Non-Hypertension 11. Similarly, studies also show that the mechanism of vascular endothelial dysfunction is associated with a decrease in plasma NO levels 6.
 
NO levels are synthesized by nitric oxide synthase (NOS) from L-arginine. After being synthesized, NO undergoes diffusion from endothelial cells into vascular smooth muscle cells and causes an increase in intracellular cyclic guanosine monophosphate (cGMP). This increase in cGMP will trigger the relaxation of vascular smooth muscle 12.
 
Nitric oxide (NO) is an Endothelial Derived Releasing Factor (EDRF) that acts as a vasodilator and lubricant to prevent the attachment of Low-Density Lipoprotein (LDL) and blood cells. Based on this study's results, in patients with hypertension, it was found that NO levels were lower compared to patients without hypertension.
 
These results are consistent with the theory that NO plays a role in regulating blood pressure and decreasing NO's availability as an essential factor during endothelial cell aging 13.

 
 
Analysis of the Effect of Consuming Pokea on Nitric Oxide Plasma Levels
 
 
In figure 1 visible difference in levels of nitric oxide (NO) in the hypertension and non-hypertensive groups where the levels of nitric oxide (NO) in the Hypertension group are lower than those of the hypertensive respondents. This is influenced by the production of depressed NO levels in hypertensive respondents.
 
NO levels play a role in blood vessel walls, precisely in endothelium vasodilation, platelet activity inhibition, and smooth muscle cells' proliferation and migration 14. If NO homo-sensitivity is disrupted by high-fat food in the blood, vasodilation will be disrupted so that the diameter of the endothelium cannot widen, causing a rise in blood pressure.
 
In the analysis of Pokea meat consumption on nitric oxide levels, it was found that there was an effect with a p-value of 0,000, while the expression of the endothelial Nitric oxide synthase (eNOS) gene also affected, with a p-value of 0,000.
 
This study seems to be in line with research (Suryana, 2017), which shows that there are significant differences in respondent's Nitric Oxide levels in the Non-Hypertension and Hypertension groups with a value of p <α (p = 0.023).
 
The mean value of Nitric Oxide levels in the Hypertension group was lower than Non-Hypertension. This is following several other studies that show that in patients with primary hypertension, endothelial-dependent vasodilation occurs due to decreased availability of Nitric Oxide 15.
 
We found higher levels of nitric oxide in people who were not hypertensive, although the average value of nitric oxide levels in hypertensive patients was above the expected value of 69,481 µmol / L; however, many hypertension respondents were below average values ​​(25–45 µmol / L), whereas in the group of not hypertension, the average value of NO was 262.8 µmol / L.
 
According to the results of the study in Padang City, showed that the majority of respondents had low levels of physical activity and lowed NO plasma with an average of 26.3 ± 15.2 µmol / L. Respondents with mild physical activity had low plasma NO levels (61.7%) compared to normal plasma NO levels (38.3%). There is a significant relationship between physical activity and plasma NO levels (p = 0.007) 16.
 
The enzyme eNOS (nitric oxide synthase enzyme) is expressed by the NOS3 gene (nitric oxide synthase-3). This enzyme plays a role in NO production, which has a vasodilating effect on blood vessels. At present, three NOS3 gene polymorphisms have been identified associated with the incidence of essential hypertension, namely Glu298Asp, -786T> C, and intron 4a4b 17.
 
Individual carriers of NOS3 gene polymorphism show decreased eNOS expression so that it can reduce NO production. NO, together with Angiotensin II, which acts as vasoconstriction, collaborates in regulating blood pressure balance. Increased Angiotensin II and decreased NO activity because continuous vasoconstriction means increased vascular resistance and blood pressure, ultimately causes hypertension 18.
 
A study conducted by Sulastri et al. in the Minangkabau ethnic community showed that the Glu298Asp allele eNOS3 gene polymorphism was not related to hypertension incidence Minangkabau ethnic group. NO plasma levels in patients with hypertension are at the lowest normal levels. Omega-3 plasma is associated with plasma NO levels only in hypertensive patients with GT heterozygous alleles 19.
 
According to the results of Sulastri et al.'s research, this supports the recommendation of omega-3 consumption in hypertensive patients with heterozygous GT alleles to increase plasma NO levels. In contrast, it is expected for normotensive sufferers to reduce the intake of saturated fatty acids because it can suppress plasma NO levels 19.
 
Nitric oxide is a compound that can convey signals to the smooth muscle in blood vessels to relax, causing blood vessel dilation (vasodilation), which results in a decrease in blood pressure. Existence of Nitric Oxide (NO) is an Endothelial Derived Releasing Factor (EDRF) that acts as a vasodilator and lubricant to prevent the attachment of Low-Density Lipoprotein (LDL) and blood cells. Nitric oxide is produced by endothelial cells from the amino origin L-arginine in a catalyzed reaction by the enzyme Nitric Oxide Synthase (NOS). Nitric oxide can cause guanylyl cyclase in vascular smooth muscle to be inactive, resulting in the accumulation of mono phosphoinositide (cGMP) guanosine and relaxation 15.
 
Nitric Oxide, also known as nitrogen monoxide, is a crucial intermediate in the body's chemical cycle. In humans, Nitric Oxide compounds are chemical compounds that are important for transporting electrical signals in cells and function in physiological and pathological processes. Likewise, this compound can cause blood vessel dilation, or in medical terms, it is called a potent vasodilator to reduce blood pressure 20.
 
Nitric Oxide (NO) plays a role in the regulation and maintenance of blood vessel pressure. NO is produced by endothelial cells and has vasodilation and antiproliferation effects on vascular smooth muscle cells. The release of NO will trigger vascular smooth muscle relaxation. A decrease in NO can occur due to a decrease in the enzyme Nitric Oxide Synthase (NOS). Decreased NOS activity causes vasoconstriction and hypertension 12.
 
Sulastri et al.'s research results, in 2011, in the Minangkabau ethnic community, showed NO plasma levels of hypertension subjects 26.91 ± 15.40 µM / L and norm tension 25.79 ± 15.04 µM / L. Thymine substitution to cytosine at position 786 was found as much as 1.5% in the hypertension group and 9.2% in the norm tension group. Low plasma NO levels (67.2%) were found in subjects with TC heterozygous alleles. There was a significant correlation between antioxidant consumption (vitamin E) and plasma NO levels in hypertensive subjects with TC heterozygous alleles (p = 0.03). Consumption of vitamin E and carotenoids can increase plasma NO levels only in patients with hypertension and normotension who have eNOS3 gene polymorphisms with TC heterozygous alleles 19.
 
In patients with hypertension, besides the ability of the eNOS3 gene to synthesize NO decreases, an unbalanced diet will worsen NO production, but diets containing high antioxidants can increase the synthesis of NO plasma. One type of food that has been reported to contain high antioxidants is Pokea.

 
 
 
CONCLUSIONS
 
Consumption of Pokea Influences NO Plasma Levels in Hypertension Patients and there are also differences in NO Plasma Levels in Patients with Hypertension and non-hypertension in Sampara District, Konawe District, South-east Sulawesi.

 
 
 
Acknowledgments
 
The author would like to thank the parties who have contributed to the implementation of this research, especially the Dean of the UHO Faculty of Medicine, Chair of the UHO LPPM

 
 
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Received: 30 November 2020
 
Accepted: 20 January 2021

 
 
 
I Putu Sudayasa1, Suryani As’ad1, Rosdiana Natsir1, Venny Hadju1, Mochammad Hatta1, Muh. Nasrum Massi1, Burhanuddin Bahar1, Sri Rahmadhani1, Yusminah Hala2, La Ode Alifariki3*
1Postgraduate Study Program in Medicine, Postgraduate Program at Hasanuddin University, Makassar, South Sulawesi, Indonesia
2Postgraduate School of Negeri Makassar University, Makassar, South Sulawesi, Indonesia
3Department of Epidemiology, Faculty of Medicine, University of Halu Oleo, Kendari, Indonesia
Corresponding author: La Ode Alifariki, Alpukat I street, Aundonohu, Kendari city, South-East Sulawesi Province, Indonesia, +6285145272116, ners_riki@yahoo.co.id
 
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