Biological Effect of Some Fast Foods Fortified with Red Chili Pepper (RCP) on Peptic Ulcer, Lipids Profile, Liver and Kidneys Functions View PDF

Fast food including spicy chicken and nuggets with red chili pepper are said to be not good for health because these foods rich in fat and cholesterol, and the high intake is associated with increased blood cholesterol levels and coronary heart disease (CHD). There is relation between red meat consumption and CHD risk factors. Dietary intake of saturated fat comes from fast food, snack food, oils and spreads, therefore in present study, biological studies were carried on selected fast foods using Sprague Dawley rats.

Chili peppers have been part of the human diet in the Americas since at least 7500 BC and perhaps earlier. There is archaeological evidence at sites located in southwestern Ecuador that chili peppers were well domesticated more than 6000 years ago [1].

In the previous century peptic ulcer was an uncommon disorder. However, during the early of the 1900s, the disease began to increase greatly. In the 1940s at least 10% to 15% of men were reported to have duodenal ulcers, with un- reported cases in women and children adding to those figures. This increasing seems to have peaked about the mid-1950s, and over the past 15 to 20 years. The knowledge on the disease process and its medical management has progressed rapidly.

Increasing of acid output, Helicobacter pylori (H. Pylori), non-steroidal anti- inflammatory drugs (NSAIDs) and stress are the basic risk factors in peptic ulcer disease [2]. Several studies found that capsaicin could have an anti-ulcer protective effect on stomach infected with H. pylori by affecting the chemicals the stomach secretes in response to infection [3-5].

The pain is the predominant symptom of peptic ulcer, although it may be absent in 25% of gastric ulcer patients. And upper gastro-intestinal hemorrhage may be the presenting sign of peptic ulcer disease, anemia from chronic blood loss, repeated vomiting and weight loss possibly happen [6].

There are many plants used for treatment of peptic ulcer, such as vegetables, fruits, and herbs. These substances had been experimented for treatment of peptic ulcer. But when we mention that spices use for healing ulcers, most people not only criticize us but also interpose with our speech. A lot of studies had been done on behalf of spices especially red chili pepper, some scientists did not find any medicinal effect of these spices on healing ulcers, as that of Myers BM, et al. (1987).

On the other hand, other scientists found that some spices especially red chili pepper has a good medicinal effect on healing ulcers. They found that red chili pepper involves functional substances called capsaicin, which has a gastro-protective effect against experimental gastric mucosal injury in animals [8-11].

Research in humans found that "after adding chili to the diet, the LDL, or bad cholesterol, actually resisted oxidation for a longer period of time, (delaying) the development of a major risk for cardiovascular disease" [12].

On the contrary, Alasdair GT, et al. (1981) remarked that hot and spicy food as long as uses aspirin is the main causes of chronic gastritis leading to ulcer disease. In this respect, red and black pepper incriminated in the increase of acid secretion [14,15].

All hot chili peppers contain phytochemicals known collectively as capsaicinoids.

• Capsaicin was shown, in laboratory settings, to cause cancer cell death in rats [16].
• Capsaicin in chilies has been found to inhibit chemically induced carcinogenesis and mutagenesis in various animal models and cell culture systems [1].
• Research on mice showed that chili (capsaicin in particular) may offer some hope of weight loss for people suffering from obesity [17].
• Researchers used capsaicin from chilies to kill nerve cells in the pancreases of mice with Type 1 diabetes, thus allowing the insulin producing cells to start producing insulin again [18].
• Research on humans found that "after adding chili to the diet, the LDL, or bad cholesterol, actually resisted oxidation for a longer period of time, [delaying] the development of a major risk for cardiovascular disease" [12].
• The amount of insulin required to lower blood sugar after a meal is reduced if the meal contains chili pepper (Blood sugar and spice Science News 2007).
• Capsaicin could have an anti-ulcer protective effect on stomachs infected with pylori by affecting the chemicals the stomach secretes in response to infection.

By combining an anesthetic with capsaicin, researchers can block pain in rat paws without causing temporary paralysis. This anesthetic may allow patients to be conscious during surgery and may also lead to the development of more effective chronic pain treatments [19].

The objectives of present work were to study:

• Biological effect of chili pepper (RCP), broast chicken and nuggets on rats with peptic ulcer.
• Biological effect of RCP, broast chicken and nuggets on fasting serum glucose, lipids profile, liver and kidneys functions for hypercholesterolemic rates.
• Histopathological effects of RCP, broast chicken and nuggets on internal organs of normal and rats with peptic ulcer.

Stages of the Work

The work of the current project is divided into 2 parts

• Biological effect of RCP, broast chicken and nuggets on rats with peptic ulcer. (included in these report).
• Biological effect of RCP, broast chicken and nuggets on hypercholesterolemic rates and hypoglycemic rates. (to be done in the next stage).

The first part (on rats with peptic ulcer)

Materials

• Plants, spicy foods: Red chili pepper (Capsicum frutescens), family were purchased from local market of KSA.
• Aspirin: Aspegic (Mmiriya Pharmaceutical Industries, Cairo) injection was prepared by dissolving one vial in 25mL distilled water to obtain solution. A volume of 1mL of this solution was orally given (at the level 200 mg/kg body weight) for one day to induce acute gastric ulcer in male albino rats.
• Diet: The rats were fed as ratio (a basal diet devoid from starch) composed of wheat bran, soya bean powder 44%, fish meal, molasses, fibers 3.3%, sodium chloride, calcium carbonate, calcium phosphate, methionine and ash (net protein 22% and fats 4.7%). The diet was fed and water was provided ad libitum for the experimental period.
• Rats: sixty-six adult male albino rats (170+5g B.Wt., each) of Sprague Dawley Strain were obtained from animal house of the Faculty of Medicine, Umm Al-Qura University (Makkah, KSA).

Preparation of Aqueous Extracts

The clean RCP (Capsicum frutescens), was ground using porcelain grinder to pass through sieve-mesh pores of 1mm diameter. The extract of red chili was prepared by mixing 1g powdered leaves with 100 mL distilled water. The mixture was boiled for 10 min. and left to cool for 15 minutes. The aqueous extract was filtered using filter paper to remove the particulate matter (0.2 mm) then the filtrate was freely dried (lyophilized) and reconstituted in 1.5 mL of distilled water (100 mg/kg body weight).

Grouping Design and Feeding of Rats

The experiment was performed in animal house of the Faculty of Medicine, Umm Al-Qura University. Rats were housed in wire cages in a room maintained at 25±2? and kept under normal healthy conditions for two weeks. All rats were fed for one week on basal diet before starting the experiment for acclimatization. After one-week period, rats were divided into two main groups. The first group (n=6 rats) was fed on the basal diet only as a control negative (healthy rats). All rats in the second main group (n=30 rats) were given orally aspirin at a dose of 200 mg/kg B.Wt., for induction of acute gastric ulcer according to Agrawal AK, et al. (2000). Rats with (aspirin-induced gastric ulcer) were disported into five groups (n=6 rats for each group) as the following:

Group 1: Control negative -ve group was fed on basal diet (non treated rats).

Group 2: Control positive +ve group fed on basal diet basal diet+Aspirin (Asp.) 200 mg/kg B.Wt.

Group 3: positive rats fed on basal diet containing 30% spicy nuggets.

Group 4: Positive rats fed on basal diet containing 30% spicy chicken.

Group 5: Positive rats fed on basal diet+oral RCP extract at doses of 300 mg/kg B.Wt.

Group 6: Positive rats fed on basal diet+oral RCP extract at doses of 600 mg/kg B.Wt.

The rats remained without food for one day prior to ether anesthesia (except for water) to avoid mixing of food with gastric secretions.

Measurement the length of gastric ulcer: At the last day of experimental period, all rats were fasted for 12-14hrs and only allowed for drinking water. In the morning of the next day, all rats were sacrificed, and their stomachs were tied around both openings (cardiac & pyloric sphincters) and injected by distilled water (3mL). The gastric juice was then collected in sterilized tube. The stomachs were opened longitudinally, washed with saline and examined under dissecting microscope for ulcer. The length of gastric ulcer was measured and expressed as mean±SD for each group. The curative ratio was then calculated for each treated group according to the method described by Akhtar AH, et al. (1995) using the following equation:

• Curative ratio (CR) = (LC - LT / LC) x 100

Where: LC = length of ulcer in control positive group

             LT = length of ulcer in treated group

Measurement the Volume of Gastric Juice

Gastric juice was collected according to the methods of Hiromichi N, et al. (1991). Abdomen was incised and both the stomach and duodenum were exposed and a fistula made by a poly ethane tube inserted into the stomach from a small incision made in the duodenum and held in place by a ligature around pylorus. In addition, esophagus was clamped to prevent reflux and loss of the gastric mucosa in tubes and centrifuged at 500 rpm for 5 min. The volume of gastric juice was measured by graduated cylinder and expressed as mL.

Histopathological Study

Specimens from stomachs were collected from rats of all experimental groups at the end of the experimental period, fixed in 10% neutral buffered formalin (pH=7.0), dehydrated in ethyl alcohol, then cleared in xylol and embedded in paraffin. 4-6 microns thickness sections prepared and stained with heamtoxylin and eosin for examining both for and glandular parts of the stomach [23].

Statistical analysis has been achieved using statistical package for social science program [24].

The present study was designated to clear out the effect of aqueous extracts of RCP and some spicy foods on healing acute gastric ulcer induced by aspirin in rats. The parameters were length of gastric ulcer, volume of gastric juice, pH, and histopathological examination of fore and glandular parts of stomachs. The effect of spicy nuggets, spicy chicken and aqueous extract of RCP (Capsium frutescens) at two doses on the length of gastric ulcer in rats were shown. The obtained results showed that:

• A single oral administration of aspirin (200 mg/kg B.Wt.) induced gastric ulcer in rats.
• The mean length of gastric ulcer in control +ve group was higher compared with in control -ve group (normal rats) being 7.27±093 and zero mm, respectively.
• Oral administration of RCP extract at a dose of 600 mg/kg B.Wt., for 7 days after aspirin (200 mg/kg B.Wt.) caused high decrease in the length of gastric ulcer (1.71±0584 mm). On the other hand, the lowest decrease of gastric ulcer length was happened in rats fed on 30% spicy nuggets, which was 4.35±0.088 mm.
• The curative ratios of ulcerated rats with oral administration of RCP extract at a dose of 600 and 300 mg/kg B.Wt., 30% spicy chicken and 30% spicy nuggets were 76.48, 71.14, 43.19 and 40.17 respectively. The highest curative ratio obtained in a dose of 600 mg/kg B.Wt., of RCP.
• The volume of gastric juice obtained from rats given aspirin-induced gastric ulcer (control +ve group) at a dose of 200 mg/kg B.Wt., was 0.69±008mL (P<0.001) compared with 0.23±0.005 mL in normal rats (control -ve group).
• Oral administration with RCP extracts at a dose of 600 mg/kg B.Wt. for 7 days after given aspirin (200 mg/kg B.Wt.) caused the highest decrease in the volume of gastric juice in rats, but oral administration with 30% spicy nuggets for 7 days after given aspirin (200 mg/kg B.Wt.) caused the lowest decrease in the volume of gastric juice in rats. The gradual ascending data were 0.31±0.004 mL (P<0.001), 0.34±0.006 mL (P<0.001), 0.39±0.010 mL (P<0.001) and 0.41±0.008 mL (P<0.01) for 600 and 300 mg/kg B.wt. of RCP and 30% spicy chicken and nuggets compared with +ve 0.69±008mL respectively.
• Effect of spicy nuggets, spicy chicken and aqueous extract of RCP (Capsium frutescens) at two doses on the pH of gastric juice collected from stomachs of rats. It is clear from data illustrated that oral administration of aspirin at a dose of 200 mg/kg B.Wt., (control +ve) decreased the pH of gastric juice (mean 3.47±07) (p<0.001) compared with (4.87±0.049) of normal rats (control -ve).
• Data showed that oral administration of RCP extract at dose of 600 and 300 mg/kg B.Wt., for 7 days after aspirin (200 mg/kg B.Wt.) reflected maximal value of pH level 4.82±0.061 and 4.02±0.060 resp. (p<0.001) compared with control (+ve). However, oral feeding of 30% spicy nuggets for 7 days after aspirin (200mg/kg B.Wt.) caused no significant decrease value (3.48±0.07).

Microscopical examination of stomach from control untreated rats revealed the normal histological structure of the stomach which consists of four layers; mucosa, submucosa, musculosa and serosa. The surface epithelial cells which are mucus secreting cells formed a continuous epithelial sheet that covered the gastric surface and line pits of gastric glands. All the cells of the surface, gastric pits and gastric glands were intact and normal without evidence of erosion or hemorrhage in the gastric mucosa. Conversely, stomach of rat from group 2 showing necrosis of the glandular mucosa associated with sloughing of laminal epithelium these superficial erosions were multiple, small and not reaching the muscularis mucosa. The sub epithelial tissues showed hemorrhage and edema with increased number of the blood vessels. There are mononuclear cells infiltrating lamina propria. Also, the stomach of this group showed destruction of gastric glands, edema in lamina propria as well as mononuclear cells infiltration. Gastric glands are reduced in number, which are abnormal in morphology and distribution. Some areas showed hyperplasic gastric glands other areas showed gastric ulceration. Covering a bier like network of hyperplasic gastric pits with destruction of upper part of gastric glands. Stomach of rat from group 3 (Asp.) +30% spicy nuggets showing destruction of gastric glands, edema in lamina propria as well as mononuclear cells infiltration. Stomach of rat from group 4 (Asp.) +30% spicy chicken showing destruction of gastric glands, edema in lamina propria as well as mononuclear cells infiltration. In Stomach of rat form group 5 (Asp.) +RCP 300 showing the normal histological structure and edema in lamina propria. There was a renewal of epithelium to line the surface of the stomach and gastric pits again, which accompanied with significant increase in the number of gastric glands. Other areas showed return of these gastric glands to the normal pattern (Tables 1-3).

Table 1: Effect of spicy nuggets, spicy chicken and aqueous extract of RCP at two doses on the length of gastric ulcer in rats.

Table 2: Effect of spicy nuggets, spicy chicken and aqueous extract of RCP at two doses on the volume of gastric juice collected from stomachs of rats.

Table 3: Effect of spicy nuggets, spicy chicken and aqueous extract of RCP at two doses on the pH of gastric juice collected from stomachs of rats.

Work Design

Hypercholesterolemia grouping: The first group (n=6 rats) was fed on the basal diet only, as a control negative (C-) (healthy rats). The second group (n=36 rats) rats was feed for two weeks on the basal diet plus cholesterol 1.5% to induce hypercholesterolemia before starting the experiment (Figures 1-14). After two weeks feeding rats divided into 4 subgroups as follows:

Group 1: Control positive group C+, was fed on basal diet plus cholesterol 2% (non treated rats).

Group 2: C+ rats fed on (1.5% cholesterol diet) diet containing RCP extract at doses of 150mg/kg B.Wt.

Group 3: C+ rats fed on (1.5% cholesterol diet) diet containing RCP extract at doses of 300 mg/kg B.Wt.

Group 4: C+ rats fed on (1.5% cholesterol diet) diet containing RCP extract at doses of 600 mg/kg B.Wt.,

Group 5: C+ rats fed on basal diet + 30% chicken.

Group 6: C+ rats fed on basal diet + 30% nuggets.

Blood sampling: At the end of the experiment (45 days), rats fasted overnight and anesthetized with chloroform. Blood samples collected be in clean dry centrifuge tubes from hepatic portal vein. Blood centrifuged for 10 minutes at 3000 rpm to separate the serum, which was kept in tubes at -18°C till analysis. Internal organs taken, washed with saline solution, and dried with filter paper, then weighted. All rats weighted once weekly. At the end of the experiment, biological evaluation of the different diets carried out by determination of body weight gain % (BWG %) and food efficiency ratio (FER) according to Chapman DG, et al. (1959) using the following formulas.

Biochemical Analysis

 Enzymatic colorimetric method was used to determine blood glucose according to Trinder P (1969). Colorimetric method was used for the determination of total cholesterol according to Allain CC, et al. (1974). Determination of HDLc were carried out according to the method of Friedewald WT (1972) and Gordon T, et al. (1977). Enzymatic colorimetric method was used to determine triglycerides according to Young D, et al. (1975). The determinations of VLDLc and LDLc were carried out according to the method of Lee R, et al. (1996) as follows: very low-density lipoprotein (VLDLc = triglycerides /5) and LDLc = (Total cholesterol – HDLc – VLDLc). Colorimetric method was used to determine AST and ALT according to Reitman S, et al. (1957); while determination of alkaline phosphates ALP activity according to Haussament TU (1977).

Effect of RCP on Organs Weight to Body Weight Ratio

 Data listed in Table 4 shows effect of RCP on organs weight to body weight ratio for control positive and different groups of hypercholesterolemic rats [34,38]. It could be noticed that the kidney and liver ratio were significant increase (P<0.05) for C +ve group rats compared with normal rats. These findings showed that there was significant increase (P<0.05) for liver weight to body weight ratio in control positive as compared to control negative group [39-41]. Hypercholesterolemic rats and fed on RCP 150 mg/kg B.Wt. and 30% nuggets showed significant (P<0.05) changes in liver weight to body weight ratio when compared to control positive [42].

Table 4: Effect of RCP on organs weight to body weight ratio for control positive and different groups of hypercholesterolemic rats.

Effect of RCP on Lipid Profile (mg/dl)

Table 5 shows that, the effect of RCP on lipid profile (mg/dl) for control positive and different groups of hypercholesterolemic rats. The results revealed that there was a significant decrease in lipid profile for control negative compared with control positive. Meanwhile, there was a significant decrease (P<0.01) in HDL of experimental animals fed on RCP 150 mg/kg B.Wt. and 30% nuggets when compared to positive group [43-48].

Table 5: Effect of RCP on lipid profile (mg/dl) for control positive and different groups of hypercholesterolemic rats.

Effect of Red Chills Pepper Extract on Atherogenic Indices for Hypercholesterolemic Rats

Atherogenic indices of control positive and different groups of hypercholesterolemic rats fed on RCP, broast chicken and nuggets were recorded (Table 6). Hypercholesterolemic rats (control positive groups) showed significant increase (P<0.01) in LDL/HDL ratio, while showed significant decrease (P<0.05) in T.C/LDL ratio and showed high significant increase (P<0.01) in TC/HDL ratio when compared to normal rats. In LDL/HDL ratio there were significant decrease (P<0.05) in RCP 600 mg/kg B.Wt while showed non-significant increase in TC/LDL ratio. At the same time, there were non-significant changes in breast chicken and nuggets when compared to control positive groups in neither LDL/HDL nor T.C/LDL ratio [49,50].

Table 6: Atherogenic indices of for control positive and different groups of hypercholesterolemic rats.

Effect of RCP on Serum Glucose (mg/dl)

Data in the Table 7 show the effect of RCP on serum glucose (mg/dl) for control positive and different groups of hypercholesterolemic rats. It is evident that the RCP 150, 300 and 600 mg/kg B.Wt. showed very slight decrease of the level of serum glucose. The mean values were (110.5, 102.5 and 107.9 mg/dl) respectively. Consequently, there were no significant difference in RCP 150, 300 and 600 mg /kg B.Wt. when compared to control positive. It was found that significant increase (P<0.05) of erum glucose could be noticed in the 30% nuggets treatment (123.8 mg/dl) compared to control positive group and the lower in 112.2 treatment (112.2 mg/dl) (Figures 15-18).

Table 7: Effect of RCP on serum glucose (mg/dl) for control positive and different groups of hypercholesterolemic rats.

• Nutritional and health educational programs should be organized and directed for the public to protect themselves from peptic ulcer and its complications.
• Spices and herbs have an effective ingredient, which can protect from ulcers, so we should add them to the meal as it is possible.
• Patients with ulcers can use red chili pepper for healing the disease, which might be used as a medicine herbs or add to food with the same percentage.
• The plants had been experimented for rats can also be used for patients with ulcer in hospitals and evaluate their nutritional states.
• Further studies should be carried out to investigate the influence of red chili pepper on their ability in correcting peptic ulcer.
• Restrict the consumption of spice chicken nuggets and spice chicken prost due to high serum cholesterol, triglycerides, LDL, and VLDL.
• While no restriction of chicken products to healthy people is not inevitable, they are advised to consume only reasonable amounts at reasonable frequencies, to avoid at least the risk of obesity, taking into consideration that many fast foods are high fat, high calories, high cholesterol and of high NaCl content.

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