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ORIGINAL ARTICLE
Year : 2022  |  Volume : 13  |  Issue : 4  |  Page : 296-300  

Ethanolic extract of Lannea coromandelica stem bark: Histopathology and infiltration of inflammatory cells in the rat's gastric


Department of Biology Education, Faculty of Teacher Training and Education, Tadulako University, Palu, Indonesia

Date of Submission04-Jul-2022
Date of Decision25-Jul-2022
Date of Acceptance08-Aug-2022
Date of Web Publication10-Oct-2022

Correspondence Address:
Dr. Achmad Ramadhan
Soekarno-Hatta Km. 9. City of Palu, Sulawesi Tengah
Indonesia
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/japtr.japtr_475_22

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  Abstract 


This study determined the efficacy of extract of the stem bark of Lannea coromandelica (ESBLc) on histopathology and inflammatory cell infiltration in the gastric of rats induced by mefenamic acid. We grouped 20 rats (Rattus norvegicus) into 5; Group 1 (positive control, mefenamic acid + sucralfate suspension), Group 2 (negative control, mefenamic acid), Group 3 (mefenamic acid + ESBLc 1575 mg/kg), Group 4 (mefenamic acid + ESBLc 3150 mg/kg), and Group 5 (mefenamic acid + ESBLc 3600 mg/kg). The dose of mefenamic acid used was 23.25 mg/kg, given orally for 7 days. Gastric histopathological observations were carried out qualitatively, and inflammatory cell infiltration was analyzed quantitatively by one-ay ANOVA. The qualitative and quantitative analysis results showed that ESBLc had efficacy in restoring damaged gastric tissue of rats; statistically, 3150 mg/kg and 6300 mg/kg effectively reduced inflammatory cell infiltration. ESBLc recovered the function of gastric organs of Rattus norvegicus L. induced by mefenamic acid, including improved mucosa and reduced inflammatory cell infiltration in the gastric. The doses of ESBLc, which effectively reduced inflammatory cell infiltrations, were 3150 mg/kg and 6300 mg/kg BW.

Keywords: Ethanolic extract, gastric, histopathology, inflammatory, Lannea coromandelica


How to cite this article:
Ramadhan A, Herman H, Sutrisnawati S. Ethanolic extract of Lannea coromandelica stem bark: Histopathology and infiltration of inflammatory cells in the rat's gastric. J Adv Pharm Technol Res 2022;13:296-300

How to cite this URL:
Ramadhan A, Herman H, Sutrisnawati S. Ethanolic extract of Lannea coromandelica stem bark: Histopathology and infiltration of inflammatory cells in the rat's gastric. J Adv Pharm Technol Res [serial online] 2022 [cited 2022 Nov 28];13:296-300. Available from: https://www.japtr.org/text.asp?2022/13/4/296/358222




  Introduction Top


Various steroidal and nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly implemented in treating inflammatory diseases,[1] including mefenamic acid NSAIDs. About 7 days of intervention of 23.25 mg/day may cause gastric abnormalities in the form of acute inflammation and erosion of the gastric mucosal epithelium in rats.[2] Treatment of stomach inflammation usually uses drugs from the antacid class; H2 antagonist drugs work by reducing the production of stomach acid. Examples of ulcer drugs included in this drug class are cimetidine, famotidine, ranitidine, sucralfate, and several other drugs.[3]

Some currently available agents used to treat ulcers have not given the desired results, and their usefulness is limited due to many side effects. Therefore, alternative control methods are needed.[4] Plant-based products seem promising in the new quest for a better ulcer treatment. Several medicinal plants reported having anti-ulcer properties were tested on experimental animals, including Musa paradisiaca, Loranthus micranthus, and Acalypha wilkesiana.[5] Lannea coromandelica belongs to the Anacardiaceae family and includes tropical trees, widely distributed in tropical countries, including India and Bangladesh,[6] conventionally various parts of this plant are used by several ethnic communities in Central Sulawesi Province, Indonesia as a medicinal plant. The bark is beneficial for treating bruises, wounds, gout, ophthalmic ulcers, ulcerative stomatitis, sprains, odontalgia, dysentery, and diarrhea.[7] In addition, the decoction of plant leaves was found effective as cough medicine, ulcer medicine, and appetite enhancer.[8]

L. coromandelica plants contain several phytochemical compounds, including saponins and flavonoids.[9] The ethanolic extract of the bark of Java (L. coromandelica) contains secondary metabolites of flavonoids, carbohydrates, gums, mucilages, and tannins.[10] The chemical content of the bark of L. coromandelica contained 13 compounds identified as quercetin, including aralia cerebroside, (2S,3S,4R,10E)-2-[(2'R)-2′-hydroxytetracosanoyl amino]-10-octadecene-1, 3, 4-triol,-sitosterol palmitate, sitosteryl-3β-glucopyranoside-6′-O-palmitate, 5,5′-dibuthoxy-2,2′-bifuran, protocatechuic acid, myricadiol, isovanillin, p-hydroxybenzoic acidethyl ester, stearic acid, palmitic acid, and trans-cinnamic acid.[11]

This study determined the efficacy of extract of the stem bark of L. coromandelica (ESBLc) on histopathology and inflammatory cell infiltration in the gastric of rats induced by mefenamic acid.


  Materials and Methods Top


The bark of L. coromandelica was obtained from the Sigi Regency, Central Sulawesi Province, Indonesia, and subsequently identified by a laboratory assistant at the Biology Education Laboratory, Tadulako University, for 3 months. A total of 500 g of dried simplicia stem bark of L. coromandelica was put into a maceration container, and 96% ethanol was added until the bark was submerged. Next, stirred and placed for 3 × 24 h and filtered to get the filtrate. Finally, the filtrate was concentrated with a rotary evaporator and weighed to calculate the yield. Phytochemical tests on the bark extract of L. coromandelica included alkaloids, saponins, polyphenols, and flavonoids.

Effects of ethanolic extract of Lannea coromandelica stem bark on rats

The white male rats of the Wistar strain, weighing 200–250 g, aged 8–10 weeks, and as many as 20 individuals were used in this study. During adaptation (7 days), they were treated and given ad libitum feed. The induction agent used to irritate the stomach was mefenamic acid 500 mg, a type of NSAID. The dose given to rats was 23.25 mg/day. These doses cause abnormalities in the stomach as acute inflammation and erosion of the gastric mucosal epithelium.[12] Mefenamic acid was administered orally for 7 days. Furthermore, the rats were orally given the ethanolic Experimental animal care and research protocols center on values and guidelines approved by the Laboratory Animal Care and Use Guidelines. We grouped the animals into 5, according to the dose [Table 1]. The selection of doses given to experimental animals was based on traditional use in the community as an ulcer reliever and has been converted to the use of experimental animals. Histological slide method of gastric organs based on the Theory and Practice of Histological Techniques guidebook.[13]
Table 1: Inflammatory cell infiltration in the stomach of experimental animals after being given extract of the stem bark of Lannea coromandelica

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Data analysis

The data from the observation of gastric histopathology were analyzed qualitatively, and the changes in gastric histology (inflammatory cell infiltration) were tested for normality with the Shapiro–Wilk test. The difference was identified using one-way ANOVA test with a 95% confidence level, followed by Duncan's multiple range test at α=0.05.


  Results Top


Phytochemicals of ethanolic extract of Lannea coromandelica stem bark

The results of the phytochemical test showed that ESBLc was positive in alkaloids, polyphenols, saponins, and flavonoids [Table 2].
Table 2: Phytochemicals of ethanol extract of Lannea coromandelica bark

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Histopathology of white rat's gastric

According to the gastric histology [Table 3] and [Figure 1], [Figure 2], there was desquamation of the gastric mucosal epithelium, mucosal erosions, and inflammatory cell infiltration with severe damage in Group 2 (negative control). Furthermore, Groups 3 and 4 had mucosal desquamation and erosion at a moderate level, but the inflammatory cells decreased. Moreover, Groups 1 and 5 experienced cell mucosal desquamation and erosion, with inflammatory cell infiltration in the mild category, which improved [Figure 1].
Table 3: The results of the qualitative evaluation of the gastric histopathological picture of white rats based on the treatment

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Figure 1: Histopathology of white rat's gastric. (a) (Group 1 positive control) reduced epithelial cell desquamation (blue arrow) and inflammatory cell infiltration (yellow arrow). (b) (Group 2 negative control) found epithelial cells desquamation and inflammatory cell infiltration. (c) (Group-3) Epithelial cell desquamation and inflammatory cell infiltration were still visible. (d) (Group-4) Epithelial cell desquamation was decreasing. (e) (Group-5) Normal epithelial cell desquamation, very less inflammatory cell infiltration (×100)

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Figure 2: Histopathology of white rat's gastric. (a) (Negative control) moderately severe mucosal erosion (black arrow). (b) (Positive control) found mucosal erosion. (c) (Group-3) Mucosal erosions are visible. (d) (Group-4) mucosal erosion was decreasing. (e) (Group-5) mucosal cell erosion began to appear normal (×100)

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White rat's gastric inflammatory cell infiltration

ESBLc had significant activity against white rats' gastric recovery induced mefenamic acid (P < 0.05). The decrease in inflammatory cell infiltration after ESBLc was given as the dose increased. The administration of 3150 and 6300 mg/Kg ESBLc effectively suppressed the amount of inflammatory cell infiltration [Table 1].


  Discussion Top


In this study, the histopathological examination of rats' gastric tissue showed that ESBLc could repair gastric tissue that experienced gastric epithelial cell desquamation, mucosal erosion, and gastric cell infiltration induced by mefenamic acid. The results showed that there were differences in gastric histopathology of Wistar rats in the negative control group (Group 2) who were not given ESBLc with the group given ESBLc (G3, G4, and G5) and with group 1 (positive control). Normal microscopic images with the gastric wall structure, such as normal mucosa, submucosa, muscular, and serous layers, were found in Group 1 and Group 5. Microscopic images of the Wistar rat's gastric in the treatment group given mefenamic acid for 7 days showed infiltration of inflammatory cells of the edema gastric mucosal stroma [Figure 1]b. Mucous irritation occurs due to increased exfoliation of mucosal epithelial cells due to drugs that irritate, thereby reducing mucus secretion, a protective barrier against acidic substances, such as gastric acid. Mefenamic acid can irritate the gastric mucosa by inhibiting the biosynthesis of prostaglandins through the cyclooxygenase enzyme.[14] The mechanism of action of mefenamic acid, like other NSAIDs, is not fully understood, but involves inhibition of cyclooxygenases (COX-1 and COX-2). Mefenamic acid inhibits both isoforms of the cyclooxygenase enzyme (COX-1 and COX-2). This prevents the formation of prostaglandins, a key in protecting the integrity of the gastric mucosa by increasing local blood flow and promoting the synthesis and secretion of mucus and bicarbonate.[15] Decreased synthesis of prostaglandins causes a decrease in mucus and bicarbonate secretion, which results in damage to the gastric mucosa. In addition, prostaglandins are vasodilators that affect widening blood vessel walls to increase blood flow to tissues. Inhibiting prostaglandins can reduce the flow of blood circulation, one of which is to the stomach. Gastric tissue can experience ischemia, which can cause the mucosa to erode.[16]

NSAIDs' anti-inflammatory and analgesic effects are based on the inhibition of cyclooxygenase, thereby inhibiting prostaglandin synthesis.[17] It may decrease HCO3-secretion, weaken mucosal protection, and stop the inhibition of acid secretion. It also damages the mucosa locally through nonionic diffusion into the mucosal cells; hence, the inhibitory effect on platelet aggregation will increase the risk of bleeding. However, further research is still needed to determine the levels of prostaglandins in the blood of rats given ESBLc.

The progression of gastric tissue repair in Group 4 and Group 5 showed that ESBLc could repair gastric tissue. The improvement was due to the content of ESBLc, which is rich in phytonutrients. Based on qualitative phytochemical tests, the ethanolic ESBLc revealed alkaloids, polyphenols, saponins, and flavonoids. Several other studies have reported that these compounds positively affect their antioxidant properties.[18] This phytochemical agent has yielded positive results when tested for its anti-ulcer and gastroprotective properties. The presence of significant phytoconstituents in the ESBLc makes it a potential candidate for future investigations.

Based on the one-way ANOVA test, the infiltration of inflammatory cells in the stomach of rats given ESBLc induced by mefenamic acid showed a significant difference (P < 0.05) among the five treatments above. The results showed that there was a significant difference (P < 0.05) between the positive control treatment (Group 1) and the negative control (Group 2) and Group 3, while the comparison between the positive control (Group 1) and Group 4 and Group 5 was not. significantly different (P < 0.05). Thus, ESBLc at a dose of 3150 mg/kg (Group 4) and a dose of 6300 mg/kg BW (Group 5) effectively reduced the amount of inflammatory cell infiltration in the stomach of white rats. With reduced cell injury, the infiltration of inflammatory cells of lymphocytes and Polymorphonuclear leukocytes (PMNs) in the gastric mucosa is reduced.[19],[20] Based on the research results, the plant understudy can be a source of new anti-ulcer. However, further research should be carried out to isolate and identify the bioactive compounds and determine their specific activities.


  Conclusion Top


ESBLc recovered the function of gastric organs of white rats (Rattus norvegicus L.) Wistar strain induced by mefenamic acid includes improved mucosa and reduced inflammatory cell infiltration in the gastric. The doses of ESBLc, which effectively reduced inflammatory cell infiltrations, were 3150 mg/kg and 6300 mg/kg BW.

Acknowledgment

The authors respectfully thank the Biology Education Laboratory, Tadulako University, for technical support and test animals.

Ethical approvals

Prior approval to conduct animal experiments was obtained from the Ethics Committee of Tadulako University (No. 09692A/UN28.1.31/PT/2020).

Financial support and sponsorship

Universitas Tadulako.

Conflicts of interest

There are no conflicts of interest.



 
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    Figures

  [Figure 1], [Figure 2]
 
 
    Tables

  [Table 1], [Table 2], [Table 3]



 

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