THE EFFECT OF PLASMA DIETARY CONTROL OF AKALINE PHOSPHATASE AND PROTEIN LEVEL IN RATS

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THE EFFECT OF PLASMA DIETARY CONTROL OF AKALINE PHOSPHATASE AND PROTEIN LEVEL IN RATS

ABSTRACT

This project research estimated the effect of Acid-base diet on plasma concentration (Alkaline Phosphatase and total protein level) in rat. The rats were induced with acidosis and alkalosis by supplementation of their diets. The level of Alkaline phosphatase and total protein were determined using colorimetric method and selectra-proS automated machine respectively, the results obtained for the ALP from the Acidotic groups were 334.7 ± 5.91, 294.2 ± 3.57, 436.2 ± 16.10 and those induced alkalosis were 209 ± 2.50, 162.1 ± 3.04 and 302.8 ± 2.80 and that of a control group is 198.7 ± 1.37 all measured in iu/L. the result of total protein obtained are 7.51 ± 0.17, 6.87 ± 0.25 and 6.47 ± 0.15 for the group induced acidosis and 6.43 ± 0.17, 7.76 ± 0.17 all measured in g/dL for the control group. The ALP level of the group supplemented with acidifying agent increases above normal level when compared to the normal range. And the group supplemented with alkalizing agent increases too but the values are within the normal range (60 – 270 ui/L ). The total protein level in groups induced with acidosis compared to control shows no significant difference (p>0.05) and so also the group induced with alkalosis but all the value are within the normal range (6.0-8.0g/dL). Therefore composition of diet can strongly affect acid and base balance thereby elevating ALP level in plasma and does not significantly affect total protein level.

CHAPTER ONE

1.0       Introduction

1.1       Background of the study

Life on earth depends on appropriate pH levels in and around living organism and cells. Human life requires a lightly controlled pH level in the serum of about 7.4 (a slightly alkaline range of 7.35 to 7.45 to survive (Waugh et al 2007).

As comparism, in the past 100 years with increasing industrialization; the pH of the ocean was dropped from 8.2 to 8.1 because of increase in carbon dioxide deposition. This has a negative compact on life in the ocean as described by (Waugh et al 2007) and (University, Brimingham 2010) and may lead to the collapse of the coral reefs confirmed (Hoegh-Guldberg, et al., 2007). Even the pH of the soil in which plants are grown can have considerable influence on the mineral content of the food we eat (as minerals are used as buffers to maintain pH. The ideal pH of soil for the best overall availability of essential nutrients is between 6 and 7. Acidic soil below pH of 6 may have reduced calcium and magnesium and soil above pH 7 may result to chemically unavailable iron, manganese, copper, zinc. Adding dolomite and  manure are ways of raising pH in an acid soil environment when the pH is below 6 (Dam-ampai, et al., 2005).

When it comes to the pH and not acid load in the human diet, there has been considerable change from the hunter-gather civilization to the present (Strohle, et al., 2010). With agricultural revolution (last 10,000) years and even more recently with industrialization (last 200 years), there has been decrease in Potassium (K) compared to Sodium (Na) and an increase in Chloride compared to bicarbonate found in the diet (Sebastain, et al., 2002).

1.2       Acid and base homeostasis

Acid-Base homeostasis is the part of human of homeostasis concerning the proper balance between acids and bases, also called pH. The body is very sensitive to its pH level, so strong mechanisms exist to maintain it outside the acceptance range of pH, proteins are denature and digested, enzymes lose their ability to function and death may occur (Caroline, et a.,l 2013).

The body’s acid-base balance is normally tightly regulated by buffering agents, the respiratory system and the renal system and the renal system, keeping the arterial blood pH between 7.38 and 7.42 (Medline Plus and Caroline, Nancy 2013).

Several buffering agents that reversibly bind hydrogen ions and impede any change in pH exist. Extracellular buffers include bicarbonate and Ammonia, whereas Proteins and Phosphate act as intercellular buffers, the relationship between multiple buffers in the same solution is described by the Isohydric principle. The bicarbonate buffering system is especially key as carbon dioxide (CO₂) can be shifted through carbonic acid (H₂CO₃) to hydrogen ions and bicarbonate (HCO^–₃) as shown below

H₂O + CO₂                       H₂CO₃                               H⁺ + HCO^–₃

(Garrett, et al.,2010)

Acid-Base imbalance that overcomes the buffer system can be compensated in the short term by changing the rate of ventilation. This alters the concentration of carbon dioxide in blood, shifting the above reaction according to Le Chatelier’s principle, which in turn alters the pH. For instance, it the blood pH drops too low (acidemia), the body will compensate by increasing breathing (medical encyclopedia metabolic acidosis) thereby expelling CO₂ and shifting the above reaction to the left such that fewer hydrogen ions are free, thus the pH will rise back to normal for alkaline, the opposite occurs. (Waugh, et a.,l 2007).

The kidneys are shower to compensate, but renal physiology has several powerful mechanisms to control pH by the excretion of excess acid or base. In response to acidosis, tubular cells reabsorb more bicarbonate from the tabular fluid, collecting duct cells secret more hydrogen and generate more bicarbonate, and Ammoniagenesis leads to increased formation of the NH₃ buffer. In responses to alkalosis, the kidney may excrete more bicarbonate by decreasing hydrogen ion secretion from the tabular epithelical cells, and lowering rate of glutamine metabolism and ammonia excretion (caroline,et al.,2013).

1.3       Acid – base imbalance

Acid – base imbalance occurs when a significant result causes the blood pH to shift out of the normal range (7.35 – 7.45). in fetus, the normal range differs based on which umbilical vessel is sampled (Umbilical vein pH is normally 7.25 – 7.45; umbilical artery pH is normally (7.18 – 7.38) an excess of acid in the blood is called alkaline or base in the blood is called alkalemia. The process that causes the imbalance is classified based on the etiology of the disturbance (respiratory or alkalosis). These are four basic processes. Metabolic acidosis, respiratory acidosis, metabolic alkalosis and respiratory alkalosis one or a combination may occur at any given time (Yeomans et al., 1985).

1.4       History

The role of the diet and its influence on the acidity of urine has been studied for decades, as physiologists have studied the kidney’s role in the body as regulatory mechanisms for controlling the acidity of body fluids. The French biologist Claude Bernard provided the classical observation of this effect when he found that changing the diet of rabbits from an herbivore (mainly plant) diet to a carnivore (mainly meat) diet changed the urine from more alkaline to more acid. Spurred by these observations, subsequent investigations focused on the chemical properties and acidity of constituents of the remains of foods combusted in a bomb calorimeter, described as ash the “dietary ash hypothesis” proposed that these foods, when metabolized, would leave a similar “acid ash” or “alkaline ash” in the body as those oxidized in combustion (Dwyer , et al., 1985).

Nutrition scientists began to refine this hypothesis in the early 20^th century, emphasizing the role of negatively charged particles (anions) in food. Diets high in chloride, phosphates and sulfates (all of which are anions) were presumed to be acid forming while diets high in potassium, calcium and magnesium (all of which are cations) were presumed to be alkaline forming. Other investigations showed specific foods, such as cranberries, prunes and plums had unusual effects on urine pH. While these foods provided an alkaline ash in the laboratory, they contained a weak organic acid, hippuric acid, which caused the urine to become more acidic instead (Dwyer, 2008).

1.5       Aim and objectives

Aim

This study is aimed at determining the effect of dietary management on the blood pH in albino wistar rat.

Objectives of the study

• To determine the effect of diet (Alkaline-acid) on alkaline phosphatase.
• To determine the effect of acid-base diet on total protein level.

1.6       Statement of problem

Diet and its effect on blood pH is an important public health challenge worldwide. Most people in the world have little or non-dietary life style without knowing the health implication, an alkaline diet also known as the alkaline ash diet, (alkaline acid diet, acid ash diet and the acid alkaline diet). Describe a group of loosely related diets on the belief that certain foods eaten can affect the acidity and pH of bodily fluids including the urine or blood, the diet and acid-base homeostasis, or the regulation of acid-base status of the body has been studied for decades, though the medical application of this hypothesis have largely focused on changing the acidity of urine, traditionally. This diet has advocated for avoiding meat, poultry, cheese, and grains in order to make urine more alkaline to prevent urinary tracts infections and kidney stones (nephrolithiasis). Acid ash has been considered as a risk factor for osteoporosis (Vangsness, et a.,l 2014).

Various risk factors that have been implicated in etiology (study of causation or origination) of blood pH is nutritional factor, poor nutritional lifestyle can cause accumulation of acid or alkaline in the blood which leads to acidemia or alkalemia respectively and also can leads to other health illness or problem, in this study, will show how diet affects the body homeostasis or blood pH, which may leads to increase or decrease in ALP and total serum protein thereby causing the body buffering system to over work itself which may cause the body system to illness health or certain diseases and causing an imbalance of the body fluids.

1.7       Scope of study

The study described in this project deal with toxicological implications of diet in blood pH in rat in other word looking into dietary modulation of the acid-base balance in rat. For this purpose the effects of a dietary load of acid or base induced changes is considered in this study and the effect of disturbances in the acid-base balance on plasma parameter routinely used in toxicity studies, to fill in this gap in knowledge, short and long-term toxicity studies were performed in rat feel with acidifying and alkalizing diets. And how these diet causes plasma parameters such as alkaline phosphate and total protein normality or abnormality in the blood.

1.8       Significance of study

The study of dietary management is very important in the sense that it enables one to identify the effect of acid-base forming diets on physiological pH, since many foods are alkaline-producing or neutral in nature but manufactured, processed foods are mostly acid producing, in other to maintain good health what one eat matters too, the body maintain correct pH in the blood at all cost by homeostasis but that is stressful for the body’s system and resources when diet is imbalance in terms of acid forming foods. These changes in alkalinity or acidity are caused by diet and lifestyle.

1.9       Limitation of the study

In carrying out this research, the researcher encountered some challenges. Urine pH, autopsy of the brain and NH₄⁺ concentration was supposed to be achieved but due to the problem encountered the researcher failed to achieve this. Some of the constraints encountered in the course of carrying out this research include;

• Lack of adequate finance
• Time constraints
• Inadequate materials in the library
• Also the epileptic power supply and
• Inconsistent availability of network posed problem. But the researcher was to gather enough material to justify the validity of the research.

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