Chinese Journal of Physiology

ORIGINAL ARTICLE
Year
: 2020  |  Volume : 63  |  Issue : 4  |  Page : 156--162

Role of dietary maize formulations in the healing of experimental acetic acid induced ulcerative colitis in male rats


Serah Funke Ige1, Mayowa J Adeniyi2, Alabi Timilehin Olayinka1, Idowu Christiana Kehinde1,  
1 Department of Physiology, Faculty of Basic Medical Sciences, College of Health Sciences, Ladoke Akintola University of Technology, Ogbomosho, Oyo State, Nigeria
2 Department of Physiology, Faculty of Basic Medical Sciences, College of Medical Sciences, Edo University, Iyamho, Edo State, Nigeria

Correspondence Address:
Dr. Serah Funke Ige
Department of Physiology, Faculty of Basic Medical Sciences, College of Health Sciences, Ladoke Akintola University of Technology, Ogbomosho, Oyo State
Nigeria

Abstract

Dietary factors do not only contribute to remission of diseases but also play important roles in the progression of medical conditions. We investigated the effect of different formulations of maize diets on the healing of experimental acetic acid-induced ulcerative colitis (UC) in male rats. Thirty-five (35) male Wistar rats (150–170 g) were randomly divided into control (CTR), UC, UC + high maize diet (HMD), UC + low maize diet (LMD), and UC + maize-free diet (MFD) groups. CTR, UC, UC + HMD, UC + LMD, and UC + MFD groups were administered different formulations of dietary maize ranging from 0% to 70%. Body weight change (BWC), colon weight, macroscopic ulcer score, catalase, glutathione (GSH), tumor necrosis factor-α (TNF-α), myeloperoxidase, diarrhea score, superoxide dismutase (SOD), Ki-67 expression, and histological studies were done. Results were analyzed using SPSS 23. UC + LMD and UC + MFD groups showed a duration-dependent reduction in negative BWC, respectively. When compared with UC group, UC + LMD and UC + MFD significantly increased (P < 0.05) GSH and SOD respectively but had no effect on TNF-α and diarrhea score. UC + HMD increased diarrhea and macroscopic ulcer scores with Ki-67 expression highest in UC + MFD. The study indicated that consumption of either LMD or maize-free diet by colitic rats relatively enhanced healing of UC.



How to cite this article:
Ige SF, Adeniyi MJ, Olayinka AT, Kehinde IC. Role of dietary maize formulations in the healing of experimental acetic acid induced ulcerative colitis in male rats.Chin J Physiol 2020;63:156-162


How to cite this URL:
Ige SF, Adeniyi MJ, Olayinka AT, Kehinde IC. Role of dietary maize formulations in the healing of experimental acetic acid induced ulcerative colitis in male rats. Chin J Physiol [serial online] 2020 [cited 2020 Sep 21 ];63:156-162
Available from: http://www.cjphysiology.org/text.asp?2020/63/4/156/293583


Full Text

 Introduction



In addition to diarrhea, ulcerative colitis (UC) is characterized by oxidative stress, increase in ulcer score and high negative body weight change (BWC).[1],[2],[3] Other markers of UC include increase in positive acute phase protein such as C-reactive protein,[4],[5] transferrin, ceruloplasmin, sialic acid, and α1-antitrypsin and increased expression of interleukins 6, 8, 1β, transforming growth factors β and tumor necrosis factor-α (TNF-α).[6],[7]

Studies on UC healing in experimental models indicated a reduction in ulcer score through stimulation of angiogenesis,[8] decrease in inflammatory, and oxidative markers such as TNF-α,[9] myeloperoxidase (MPO), and reactive oxygen species and reduction in diarrhea score.[10]

Furthermore, many factors have been implicated in UC healing. For example, green tea polyphenol and Garciniakola crude extract have been shown to accelerate colitis recovery by improving antioxidant levels.[11],[12],[13] Dietary milk fat and high cholesterol diet were separately reported to aggravate colitis through the attenuation of colon unfolded protein response.[11],[14],[15] Daily consumption of meat and sugar has been associated with an increased incidence of UC in human studies.[16]

Conversely, animals fed fat-free diet recovered faster from experimental colitis.[17] Supplementation with probiotics and prebiotics elicited remission of colitis.[18] High fiber-containing foods have been extensively reported to improve UC healing due to their anti-inflammatory effect[16] with attendant facilitation of bowel movement.[19] Despite the fact that maize is one of the staple foods, its effect on UC healing is not well understood. The present study aimed at determining the effect of different formulations of dietary maize on UC healing using rat model.

 Materials and Methods



Animal care and management

Thirty-five male Wistar rats used for this research work were obtained from the Animal House, Department of Physiology, Ladoke Akintola University of Technology, Ogbomoso, Oyo State. They were kept in a plastic cage (37 cm × 27 cm) and lined with wood shavings. The animals were maintained in 12-h light/12-h dark cycle at room temperature.

Ethical certification

The study was conducted in line with the guidelines of National Institute of Health for the use of laboratory rats.

Experimental design

The animals were randomly grouped into five groups containing seven animals each:

Group I – Were given 0.3 ml of distilled water and fed normal feed mealGroup II – Were colitic rats fed normal feed mealGroup III – Were fed high maize diet (HMD) after colitis inductionGroup IV – Were fed low maize diet (LMD) after colitis inductionGroup V – Were fed maize-free diet after colitis induction.

The animals were given different formulations of maize diet for 7 days.

Formulation of the feed

The feed was obtained and formulated at Gloryvet Livestock Feedmill, Ogbomoso, Oyo State, Nigeria [Table 1]. The feed was formulated following the method by Abdulai etal.[20] with modifications.{Table 1}

Determination of body weight change

The animals were weighed every day during the research process and the change in body weight was determined by:

BWC (g) = Final weight (g) − Initial weight (g).

Induction of colitis

After the animals were fasted for 24 h, colitis was induced in Groups II–V through a single intra-colonic instillation of 7% acetic acid (1 ml/100 g of body weight). On the other hand, the animals in Group I were administered distilled water. All animals were sacrificed through cervical dislocation and the colons of the animals were excised.

Tissues preparation

After the 7th day of induction, all the rats used for the research were weighed and euthanized by cervical dislocation. They were dissected and the distal colon was excised, weighed, and washed in phosphate buffer. Half of the tissues were cut longitudinally for histological studies and Ki-67 expression. The other half was put in a sample bottle containing phosphate buffer.

The assessment of diarrhea and macroscopic ulcer scores

Stool consistency was monitored and scored daily according to the method of Masonobi etal.[21] with modification. Macroscopic damage was assessed by the scoring system of Zheng et al.[22]

Determination of antioxidant parameters and lipid peroxidation

The level of colonic superoxide dismutase (SOD) activity was determined by the method of Mistra and Fridovich.[23] Colonic catalase activity was determined using the method of Sinha.[24] The method of Beutler etal.[25] was used in estimating the level of colonic glutathione (GSH). The content of MDA in the homogenates was determined spectrophotometric ally using the method of Varshney and Kale.[26]

Determination of tumor necrosis factor-α and myeloperoxidase

Tumor necrotic factor α and MPO were assayed using enzyme-linked immunosorbent assay kits; Quantikine Elisa and Elabscience Biotechnology Inc., respectively.

Histological analysis

The colon was processed through graded alcohols into paraffin wax. Paraffin-embedded tissues were serially sectioned at 3 μm and stained with a modified Masson trichrome stain. The photomicrographs of the colon were then obtained.

Immunohistochemical study

A section of the colon was excised and washed in 1.15% potassium chloride. The tissues were preserved in formalin for Ki-67 expression in the colon. Formalin-fixed and paraffin-embedded tissue samples of the colon were cut into 4-mm sections and dried overnight at 60°C. Representative sections were stained using hematoxylin and eosin prior to immunostaining. All the slides were examined and scored independently by the observers in blinded fashion.

Statistical analysis

Values were expressed as mean ± standard error of the mean. The comparisons were made using analysis of variance and Student's t-test. Statistical significance was accepted at P < 0.05.

 Results



Colonic weight and body weight change in colitic rats with different dietary maize formulations

The induction of UC using acetic acid caused a significant increase in colon weight when compared with control (CTR) group [Figure 1]a. In colitic rats, consumption of HMD or maize-free diet (MFD) increased colon weight significantly when compared with CTR group. When compared with colitic group, consumption of LMD by colitic rats caused a significant reduction in colon weight.{Figure 1}

Induction of UC using acetic acid caused a significant negative BWC when compared with CTR group [Figure 1]b. In colitic rats, administration of HMD, LMD or MFD significantly decreased BWC when compared with CTR. When compared with colitic group, HMD significantly increased negative BWC in UC rats 7 days' post-induction whereas LMD or MFDs significantly reduced negative BWC. There was a duration dependent decrease in negative BWC in UC rats fed LMD or MFD.

Changes in diarrhea and macroscopic ulcer scores in colitic rats with different dietary maize formulations

Induction of UC caused a significant increase in diarrhea score when compared with CTR group [Figure 2]a. Consumption of HMD, LMD or MFD by colitic rats resulted in a significant increase in diarrhea score when compared with ulcerative CTR group.{Figure 2}

Induction of UC caused a significant increase in macroscopic ulcer score when compared with CTR group [Figure 2]b. In colitic rats, consumption of LMD or MFD caused a significant reduction in macroscopic score when compared with colitic group. In colitis rats, consumption of HMD increased macroscopic ulcer score significantly when compared with colitic group.

Colonic antioxidant parameters in colitic rats fed with different dietary maize formulations

Induction of UC resulted in significant reduction in SOD [Figure 3]a. When compared with colitic group, consumption of LMD or MFD by colitic rats resulted in a significant increase in SOD.{Figure 3}

Induction of UC significantly decreased catalase [Figure 3]b. When compared with colitic group, consumption of HMD, LMD or MFD by colitic rats resulted in an increase in catalase.

When compared with colitic group, consumption of LMD or MFD by colitic rats significantly increased GSH [Figure 3]c.

Lipid peroxidation in ulcerative colitic rats fed with different dietary maize formulations

When compared with CTR, consumption of HMD, LMD or MFD by colitic rats resulted in increase in malondialdehyde [Figure 4].{Figure 4}

Inflammatory markers in ulcerative colitis rats fed with different dietary maize formulations

Induction of colitic using acetic acid caused a significant increase in MPO [Figure 5]a. When compared with colitic group, consumption of LMD by colitic rats led to a significant reduction in MPO.{Figure 5}

When compared with colitic group, consumption of LMD by colitic rats significantly decreased TNF-α [Figure 5]b.

Colonic histology of ulcerative colitis rats fed with different dietary maize formulations

CTR group showed normal mucosal layer with numerous intestinal glands [Figure 6]a. Colitis + normal diet showed focal area of mucosal ulceration, the lamina propria and submucosal layer with severe infiltration of inflammatory cells [Figure 6]b. Colitis + LMD showed normal mucosal layer with numerous intestinal glands, the lamina propria, and submucosal layer revealing moderate infiltration of inflammatory cells [Figure 6]c. Colitis + MFD showed normal mucosal layer with intestinal glands, the lamina propria, and submucosal layer with moderate infiltration of inflammatory cells [Figure 6]d. Colitis + HMD in [Figure 6]e showed acutely inflamed mucosal layer; the lamina propria, the glands and submucosal are severely infiltrated with inflammatory cells.{Figure 6}

Ki-67 expressions in ulcerative colitis rats fed with different dietary maize formulations

As shown in [Figure 7], colitis group shows moderate expression of Ki-67 within the nuclei of the mucosa glands. Colitis + LMD group in [Figure 7]c shows mild expression of Ki-67 within the nuclei of the mucosa glands and inflammatory cells. Colitis + MFD group in [Figure 7]d shows severe expression of Ki-67 within the nuclei of the mucosa glands and inflammatory cells. Colitis + HMD group in [Figure 7]e shows slightly moderate expression of Ki-67 within the nuclei of the mucosa glands and inflammatory cells.{Figure 7}

 Discussion



Preponderance of studies has indicated the pros and cons of nondrug therapy in the remission of UC, a medical condition characterized by diarrhea, weight loss, elevation of inflammatory markers, and high ulcer score.[12],[16] The present investigated the effect of dietary maize formulations on healing of UC using rat model.

The significant reduction in ulcer score of UC rats that were administered LMD or MFD might be due to stimulation of angiogenesis.[8] Evidence from the immunohistochemical study also showed an increase in expression of Ki-67, a cellular marker of cellular proliferation.[27] For example, colitic rats that were administered maize-free diet exhibited the highest Ki-67 expression. This together with relative reduction in inflammatory cells implied that mucosal healing, which is the ulterior treatment goal of UC might be better achieved through consumption of diet with little or no maize content. Over the years, some researchers have advocated for the inclusion of fiber-rich foods in dietary panel for patients with inflammatory bowel diseases such as UC despite the fact that these foods demonstrate tendency of increasing intestinal motility.[16],[19] As far as the present study was concerned, our major focus was not the fiber containing potency of maize but to clarify whether formulations of maize diet enhance colitis healing.

Besides ulceration, UC is characterized by diarrhea.[1] The result of our study confirmed diarrhea as a feature of colitis. We also noticed that all groups that were fed with different formulations of maize diets exhibited significant increase in diarrhea score, even though the highest mean diarrhea score was observed in UC rats that were fed HMD. As far as the study was concerned, the influence of fiber-enriched foods on gut microta growth and the contribution of gut microbiota to immune function and inflammation cannot be ruled out.[28] Underlying mechanism of diarrhea might also include upregulation of cystic fibrosis transmembrane regulator of chloride ion transport.[29],[30]

We observed that consumption of LMD or MFD caused a reduction in negative body weight in UC. This might be due to the facilitative influence of LMD or MFD on UC healing and relative suppressing effect of these diets on diarrhea score. The present study also indicated a duration dependent decrease in negative BWC in UC rats that were fed with LMD or MFD. This finding implies that in UC rats, as the frequency of consumption of LMD or MFD increased, there was alleviation of weight loss.

Like previous studies,[2],[12] experimental acetic acid induced colitis resulted in antioxidant dyshomeostasis evidenced by reduction in SOD and catalase. However, when compared with UC group, consumption of LMD orchestrated a significant increase in catalase, SOD and GSH. This finding indicates that the anti-ulcer effect of LMD may be attributed to improvement in antioxidant strength.

The significant reduction in inflammatory markers such as MPO and TNF-α in UC rats that were fed with LMD provides additional mechanism through which remission of UC occurs. Previous works also indicated that UC healing was characterized by decrease in inflammatory markers such as TNF-α[9] and MPO species.[10]

The present study demonstrated the relative advantage of consumption of LMD or MFD by UC rats over HMD. Our study also indicated that the relative therapeutic advantage of LMD was due to increase in SOD and catalase and reduction in inflammatory markers such as MPO and TNF-α. Furthermore, further studies may be required to clarify the role of maize diet on Chrohn's disease, another inflammatory bowel disease.

 Conclusion



The study indicated that the consumption of LMD or maize-free diet by colitic rats relatively enhanced healing of UC.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

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