|Year : 2020 | Volume
| Issue : 9 | Page : 403-410
Opuntia humifusa aqueous extract alleviates ethanol-induced gastric ulcer in a mouse model
Chi-Yeol Yoo1, Hyeong-U Son2, Alshammari Fanar1, Hee-Jung Choi1, Md Badrul Alam3, Sang-Han Lee2
1 Department of Food Science and Biotechnology, Kyungpook National University, Daegu 41566, Korea
2 Department of Food Science and Biotechnology; Food and Bio-Industry Research Institute, Kyungpook National University, Daegu 41566, Korea
3 Food and Bio-Industry Research Institute, Kyungpook National University, Daegu 41566, Korea
|Date of Submission||29-Oct-2019|
|Date of Decision||09-Dec-2019|
|Date of Acceptance||15-Apr-2020|
|Date of Web Publication||30-Jul-2020|
Department of Food Science and Biotechnology; Food and Bio-Industry Research Institute, Kyungpook National University, Daegu 41566
Source of Support: None, Conflict of Interest: None
Objective: To investigate the effect of Opuntia humifusa aqueous extract on gastric ulcers.
Methods: An ethanol-induced model was used to examine the protective effect of Opuntia humifusa against gastric ulcers. The gastric ulcer index was evaluated via clinical observation and image analysis. Various inflammatory indicators were determined by RT-PCR and Western blotting assays.
Results: The gastric ulcer index was reduced to 8% in the group treated with Opuntia humifusa aqueous extract compared with that in the control group. RT-PCR analysis revealed that MUC5AC expression was reduced to 39% in the control group compared with the non-treated group, whereas the omeprazole and Opuntia humifusa aqueous extract-treated groups increased the expression to 95% and 79%, respectively. Moreover, the expressions of various cytokines including TNF-α, IL-1β, and IL-6 were increased in the control group, while decreasing in Opuntia humifusa aqueous extract-treated group. Opuntia humifusa aqueous extract also suppressed the expressions of iNOS, COX-2, and its transcription factor NF-κB and increased mucus content considerably as compared to the control group.
Conclusions: These results suggest that Opuntia humifusa aqueous extract is suitable as an alternative remedy for gastric ulcer treatment.
Keywords: Opuntia humifusa; Gastric ulcer; Gastric lesions; Image analysis; Mucin
|How to cite this article:|
Yoo CY, Son HU, Fanar A, Choi HJ, Alam MB, Lee SH. Opuntia humifusa aqueous extract alleviates ethanol-induced gastric ulcer in a mouse model. Asian Pac J Trop Biomed 2020;10:403-10
|How to cite this URL:|
Yoo CY, Son HU, Fanar A, Choi HJ, Alam MB, Lee SH. Opuntia humifusa aqueous extract alleviates ethanol-induced gastric ulcer in a mouse model. Asian Pac J Trop Biomed [serial online] 2020 [cited 2020 Dec 5];10:403-10. Available from: https://www.apjtb.org/text.asp?2020/10/9/403/290131
| 1. Introduction|| |
Gastric ulcers have a profound impact on human health worldwide. Gastric ulcers combine to duodenal ulcers to form peptic ulcer disease, which is the most common gastrointestinal disorder.
Major factors that impact gastric ulcers are stress, alcohol, tobacco, steroids, food irritants, nutritional deficiencies, and Helicobacter pylori infection,. However, deterioration of antioxidant capacity owing to an increase in reactive oxygen species (ROS) and prostaglandin production is associated with ulcerative lesions. Acute gastric ulcers are reportedly induced by a single dose of indomethacin, ethanol, aspirin, and water immersion-restraint stress; such models have been used to monitor a variety of factors that affect gastric injury treatment, thereby demonstrating that different factors/agents can cause different types of ulcers. Moreover, gastric ulcers are treated by signal mediators, including prostaglandins, cytokines, and matrix metalloproteinases, which are needed to maintain gastrointestinal homeostasis for regulating gastrointestinal lesion development or healing mechanism.
Additionally, mucin is a major polymeric glycoprotein component of the mucosal surface (10-μm sized granules with a fiber-like membrane), which plays an important role in preserving mucosal expression integrity. It is present in various systemic secretions from the gastrointestinal mucosa, respiratory mucosa, female genital mucosa, middle ear, and conjunctiva. Moreover, mucin is a polypeptide in which oligosaccharides are linked by O-linked glycosylation, and the sugar structure of mucins has different composition ratios depending on the tissue and disease. Moreover, mucus represses cell adhesion of cancer cell growth, immune recognition, and gastric epithelial cells. In addition, the risk of a malignant transformation increases and adversely affects intrusion and metastatic ability. In these cases, steroids and non-steroidal antiinflammatory drugs are generally used for treating mild symptoms. However, systemic long-term administration may cause side effects such as immunosuppression and gastric ulcer. From this viewpoint, it is important to search for natural anti-inflammatory products whose safety has been established. Additionally, in Western medicine, omeprazole is a widely used drug for gastric ulcer treatment; human and animal experiments have revealed that omeprazole can be substituted by benzimidazole, which is used to treat gastric ulcer and gastric hyperplasia because of the latter’s strong ability to inhibit gastric acid secretion.
Opuntia humifusa (O. humifusa), a member of the Cactaceae family, is a perennial plant and widely distributed across semi-arid regions, such as Central and South America. O. humifusa has also been cultivated in large quantities in Asan and Uiseong, Republic of Korea, and is known as “Cheon-nyun-cho” in Korean. It has biological benefits such as radical scavenging and anti-inflammatory activities, exerts streptozotocin-mediated anti-diabetes effect and anti-asthmatic effect by regulating the expression levels of IL-4 and IL-13 as well as ameliorates acute pancreatitis. In particular, the plant contains mucilage, which consists of carbohydrates in the form of pectin, arabinose, galactose, galacturonic acid, and rhamnose, and can ameliorate gastric ulceration. Therefore, our research focused upon the effect of O. humifusa aqueous extract on gastric ulcers.
| 2. Materials and methods|| |
The following reagents were purchased: ethyl alcohol (99.9%, Merck), TRIZOL reagent (Invitrogen, Carlsbad, CA), omeprazole (O104, Sigma-Aldrich, St. Louis, MO), caffeic acid (C0625, Sigma- Aldrich), ferulic acid (PHR1791, Sigma-Aldrich), gallic acid (G7384, Sigma-Aldrich), chlorogenic acid (C3878, Sigma-Aldrich), quercetin (Q4951, Sigma-Aldrich), and rutin (R5143, Sigma- Aldrich).
2.2. Sample preparation
O. humifusa (1 kg) was obtained from Uiseong Cluster Association, Republic of Korea; The Association Group supervised cultivation to manage standard O. humifusa. The voucher specimen (2017- OH) was deposited in the Food Enzyme Biotechnology Laboratory, Kyungpook National University, Daegu, Korea. O. humifusa was dried and homogenized in a commercial mixer (JSHR-270D, JSR, Gongju, Korea). The partial remaining powder (100 g) was mixed with distilled water at a 1:10 w/v ratio and extracted using a shaking incubator (Korea) at 45 °C for 24 h. After extraction, the mixture was filtered using WHATMAN filter paper No. 1 and lyophilized (Lyophilizer, Ilshin BioBase, Dongducheon, Korea), after which it was dissolved in distilled water for further investigation.
2.3. Animal care
Five-week-old specific pathogen-free male BALB/c mice (average body weight, 20-23 g) were purchased from Samtaco Korea (Osan, Korea). The experimental animals were subjected to laboratory purification for 1 week in the animal breeding room of Kyungpook National University Experimental Animal Research Center. The mice were freely fed a pellet-type solid feed Samtaco Korea (Osan, Korea) and allowed to drink water freely.
2.4. Ethical statement
This experimental procedure was performed strictly according to in-house guidelines and the current protocol conformed with the Guidelines Research and Ethical Issues of the International Society of Pain Medicine. Kyungpook National University Industry Foundation (KNUIF-2017-0069).
2.5. Ethanol-induced gastric ulceration model
Ethanol-induced gastric ulceration model was performed as described previously, with minor modifications. The mice were randomly divided into four groups (n=5), namely, normal control, ethanol control (ethanol induction and saline treatment), omeprazole group (ethanol induction and omeprazole treatment, 3 mg/kg) and O. humifusa aqueous extract-treated group (ethanol induction and O. humifusa aqueous extract, 100 mg/kg via p.o. injection with 200 μL saline). The concentration of O. humifusa aqueous extract was fixed since 100 mg/kg treatment was sufficiently effective against gastric ulcer in our previous study (data not shown). Omeprazole was used as the positive control. The animals were fasted and allowed to consume only purified water for 18 h. Before ethanol induction, mice were subjected to the oral administration of each sample and maintained without water ad libitum for 30 min. All mice groups, except normal control, were orally administered with 8 mg/ kg ethanol. After 30 min, the mice were intoxicated and laid down because of ethanol induction, but no death occurred. The mice were then euthanized by CO2 gas, and subsequently, gastric tissues were isolated.
2.6. Image analysis of gastric ulcers
Using image analysis, we examined striped erythema or pointlike lesions and various damaged gastric tissues for measuring ulcer index scores. Photographed image was divided into ulcer lesion or none because of its distinct color. Ulcer area was calculated as selected pixel by Image J program. To maintain the consistency of the measured images, we controlled the light source and the digital light raw file. We took pictures in a dark room using a controlled box in a restricted space to avoid environmental disturbances caused by sunlight (Hyundai Fomax E400, Seoul, Korea). A Nikon digital camera was used (D5100; Nikon Co., Tokyo, Japan). The Nikon digital camera D5100 was equipped with AF-S Micro NIKKOR 40 mm f/2.8 G lens (Nikon Co., Tokyo, Japan) and a Nikon ML-L3 wireless remote-control system (Nikon Co., Tokyo, Japan). In the first experiment, the subject was placed in a small studio with a diffuser and a background fabric cover, followed by a color image scale (QP card 203, QPcard AB, Helsingborg, Sweden). Color images are innovative tools that control color profiles and enable color reproduction. All photograph data were conducted on same condition for calculating gastric lesions area. Images of gastric mucosal organs were saved as original files and colors were adjusted using a QP Calibration v1.21 software (QPcard AB, Helsingborg, Sweden).
2.7. Measurement of gene expression (MUC5AC, TNF-α, IL-6, IL-1β, COX-2, GAPDH) by RT-PCR
Total cellular RNA was prepared using TRIZOL® reagent according to the manufacturer’s instruction. After mice were euthanized, gastric tissues were isolated and the total RNA was extracted as described. One mL of TRIZOL® reagent was mixed with frozen stomach tissues (approximately 100 mg). The total RNA content was analyzed by projecting a multi-label counter (VICTOR-3 spectrophotometer, Perkin Elmer, Turku, Finland) at 260 nm. After the course of the reaction, each product was analyzed using 1% agarose gel electrophoresis by performing EtBr staining. The gene product was identified by comparing with the 1-kb DNA ladder (Solgent in Korea). For semi-quantitative analysis, the relative intensities of each set of gene products were quantified using a Molecular Imager (ChemiDoc XRS+, Bio-Rad, Philadelphia, PA, USA). PCR reaction was carried out in a programmable thermal cycler (Perkin Elmer, model 9700).
2.8. Immunohistochemical analysis
Immunohistochemical analysis was performed as described in the previous experiment . Briefly, the stomach tissue was cut into larger curvatures and embedded in paraffin. Thereafter, the section was cut to a size of 5 mm and placed on a slide warmer for 16 h. Xylene was used for deparaffinization, after which each section was serially diluted with 70%–100% alcohol for 60 min. After washing, the sections were incubated with 10% goat serum at RT for 1 h to avoid non-specific binding. The sections were subsequently stained with antibody (iNos and NF-κB).
2.9. HPLC analysis
Experiments were performed on a Shimadzu Prominence HPLC system (Shimadzu, Kyoto, Japan). The extract samples were eluted at a rate of 10 mg/mL and filtered through a syringe filter (0.2 μm, Pall Life Sciences, Ann Arbor, MI). A C18 column (4.6 × 250 mm, 5 μm) was used, with a flow rate and column temperature of 1 mL/min and 30 °C, respectively; the detection wavelengths were set at 254 and 324 nm. The mobile phase was 1% formic acid: MeOH at a ratio of 80:20.
2.10. Statistical analysis
Data are expressed as mean ± standard deviation (SD). Statistical values were calculated by ANOVA with Tukey post-hoc test program using SPSS 21.0 software. P<0.05 was considered statistically significant.
| 3. Results|| |
3.1. Effect of O. humifusa aqueous extract on gastric ulcer
Severe lesions in the stomach were phenotypically confirmed. As shown in [Figure 1]A, gastric ulcer lesions resulted in a light peach to red coloration in the inner tissues lining the stomach. Red-colored areas were clearly seen in the images, demonstrating tissue damage in the ethanol control group, and red-colored spots/lesions were measured in black color using Image J for alternative comparison. For easily comparing the area values, white and black colors through binary were shown [Figure 1]. Image analysis revealed that the omeprazole-treated group (positive control) showed a significant decrease in the positive area, whereas the O. humifusa aqueous extract-treated group showed the index was reduced to 8% in the positive area compared with the ethanol group [Figure 1]B. These results indicated that the activity of O. humifusa aqueous extract was similar to omeprazole.
|Figure 1: Effects of Opuntia humifusa (O. humifusa) aqueous extract on mucosal tissues in an ethanol-induced gastric ulcer model. (A) Images of gastric ulcers in the experimental groups. (B) Positive area was measured according to gastric lesion index as shown in Figure 2A and expressed as the intensity relative to control. Data are expressed as mean ± standard deviation (SD, n=5 animals/group). #P<0.05 and *P<0.01 compared with the normal control group and the ethanol-administered control group, respectively.|
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3.2. Relevant gene expression by RT-PCR
Expression of MUC5AC was decreased to 39% in the ethanol control group compared with the normal control group. Treatment with O. humifusa aqueous extract as well as omeprazole increased the expression to 79% and 95%, respectively [Figure 2]B. Additionally, the ethanol control group increased TNF-α, IL-6, IL-1β and COX-2 expressions, while omeprazole and O. humifusa aqueous extract decreased these expressions [Figure 2]C, [Figure 2]D, [Figure 2]E, [Figure 2]F. The effect of O. humifusa aqueous extract was comparable to that of omeprazole.
|Figure 2: Analyses of RT-PCR for measuring the inhibition of mRNA expression by O. humifusa aqueous extract. Comparative analysis of mRNA expression in ethanol-induced ulcer lesions after treatment with O. humifusa aqueous extract was performed using RT-PCR. (A) the detected PCR band. (B-F) the gene expressions of MUC5AC, TNF-α, IL-6, IL-1β, and COX-2 mRNAs, respectively.|
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3.3. Immunohistochemical results
The distribution of responsive molecules in stomach tissues was measured by periodic acid Schiff (PAS) staining. In the normal control group, dark pink coloration was observed on the surface of the stomach tissue sections owing to PAS staining. The control group showed a reduced number of lesions in the barrier spot [Figure 3]. For more accurate determination of the symptoms, image analysis was performed with a similar protocol as followed for measuring gastric ulcer index. A viscous molecule, such as a polysaccharide, was depicted as a black area, and the other parts were depicted as a white area [Figure 3]. Quantification of these black areas revealed a 87% reduction in the ethanol group compared with the normal control group, whereas 14% and 29% reduction were observed in the omeprazole and O. humifusa aqueous extract-treated groups, respectively, indicating a protective effect against gastric ulcer [Figure 3]. Moreover, inducible nitric oxide synthase (iNOS) expression in stomach tissues was observed after ethanol induction, although it was not observed in the normal control group. The results of the antibody-bound area were shown in [Figure 4]. Omeprazole treatment showed a 57% reduction in antibody-bound areas compared with an assumed value of 100% for the ethanol group. In contrast, O. humifusa aqueous extract reduced iNOS expression more significantly [Figure 4]. Moreover, O. humifusa aqueous extract restored NF-κB expression towards normal [Figure 5]. Image analysis further confirmed the results of quantitative analysis [Figure 5].
|Figure 3: Immunohistochemical analyses by PAS staining. Immunohistochemistry was applied to compare the protective effects of O. humifusa aqueous extract via clinical observation or image analysis. (A) Dark violet blots indicate PAS-responsive cells. (B) Relative band intensities are shown in bar graph. Significant differences (P<0.05, ANOVA with Tukey post-hoc) when compared with the normal control (#) or the ethanol control (*) group. PAS: periodic acid Schiff.|
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|Figure 4: Expression of inducible nitric oxide synthase (iNOS) by immunohistochemical analysis. (A) Dark violet blots indicate iNOS-responsive cells. (B) Relative band intensities are shown in bar graph. Significant differences (P<0.05, ANOVA with Tukey post-hoc) when compared with the normal control (#) or the ethanol control (*) group.|
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|Figure 5: Expression of NF-κB by immunohistochemical analysis. (A) Dark violet blots indicate NF-κB-responsive cells. (B) Relative band intensities are shown in bar graph. Significant differences (P<0.05, ANOVA with Tukey post-hoc) when compared with the normal control (#) or the ethanol control (*) group.|
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3.4. HPLC analysis of O. humifusa aqueous extract
O. humifusa aqueous extract was analyzed for identifying the compounds by HPLC analysis [Figure 6]. The active components present in the O. humifusa aqueous extract included 384.1 μg/g gallic acid, 258.8 μg/g rutin, 147.3 μg/g chlorogenic acid, and 503.5 μg/g ferulic acid. However, quercetin and caffeic acid were not detected.
|Figure 6: HPLC analysis of O. humifusa aqueous extract. Standards (red line) were clearly separated and the sample (blue line) was loaded and compared with standards. (1) gallic acid, (2) chlorogenic acid, (3) caffeic acid, (4) ferulic acid, (5) rutin, and (6) quercetin.|
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| 4. Discussion|| |
We evaluated the gastroprotective effects of O. humifusa aqueous extract on an ethanol-induced animal model. The plant is well-known to contain higher concentrations of polyphenolics, flavonoids, and dietary fibers than any other subtropical plants. The extracts of O. humifusa exhibit anti-inflammatory,, anti-diabetic, anti-aging, skin whitening, anticancer, and antioxidative effects. However, the anti-ulcer effects of O. humifusa remain unclear. Therefore, we investigated the gastroprotective ability of this plant extract using an ulcer model.
Contrastingly, it has been well-documented that MUC5AC encodes mucin in human beings. A part of the conformational structure in MUC1 and MUC6 consists of small recurring fragments of amino acids; consequently, the core region of mucin undergoes structural modification, which in turn substantially modifies its function. Out of the nine investigated genes, so far, chromosome 11 was found as the locus of MUC5AC, MUC5B, and MUC6. Moreover, TNF-a, a very well-known cytokine that causes systematic inflammation, is the functional modulator of gastric mucosal lesions. Acute inflammation reaction caused gastric ulceration, which was followed by neutrophil infiltration into the gastric mucosa. It has recently been revealed that gastric mucosal lesions can cause activation of the caspase-3-induced apoptotic pathway,. Preliminary experimental observations reveal that gastroprotective effects of O. humifusa are considerably predominant with the inhibition of TNF-α, IL-1β, and IL-6, whereas these proinflammatory cytokines played a significant role in ethanol-instigated gastric ulcers. TNF-α expression levels were drastically decreased with O. humifusa aqueous extract treatment. In the inactivated state, NF-κB is retained in the cytoplasm IκB protein. When the tissues are exposed to various inflammatory stimulators, IκB kinase induces IκB protein phosphorylation and causes decomposition of IκB complexes. This event of NF-κB/IκB balance transferred to the nucleus and caused transcriptional activation of other proinflammatory cytokines including TNF-α and IL-6,. The present results suggested that the downregulation of the NF-κB signaling pathway was mediated by ethanol-induced COXs, NOSs, and other inflammatory cytokines during gastric ulcer induction. Therefore, we investigated mRNA expression by examining COX-2, IL-6, IL-1β and TNF-α levels. The data unveiled that the downregulation of the NF-κB signaling pathway was mediated by ethanol-induced gastric COX isoenzymes and prostaglandins (data not shown). Moreover, COX-2 is predominant in inflamed areas. In a parallel experiment, it was concluded that regulation of gene expression between COX-1 and COX-2 as well as cytokines is closely associated with inflammation homeostasis,,. Screening of several food extracts confirmed the existence and expression of ulcer inflammatory biomarkers. COX-2 is an anti-inflammatory drug target. Moreover, IL-6 is a cytokine and is known to be a major regulator of acute and chronic inflammation as well as a stimulant of T and B cells. Cytokine stimulates neutrophils, macrophages, and lymphocytes in inflammatory sites to generate toxic products, ROS, and lysosomal enzymes that cause damage of various tissues during the development of gastric ulcer. Previous studies have demonstrated that mRNA levels of IL-6 are significantly higher in inflamed tissues of gastric ulcers. The overproduction of IL-6, IL-1β, and/or TNF-α is responsible for inflammatory disease progression, and antibody-based therapy has been shown to significantly reduce inflammation by blocking inflammatory signals.
Image analysis program is a fascinating tool for measuring phenotypic symptoms using photographic data, including immunohistochemical images, generated from computer programs, such as ImageJ, Adobe Photoshop, and Metamorph software. Because the process eliminates unnecessary information, which made it easy to confirm any differences, we can measure the individual areas of damaged tissues by concurrently estimating binary viewing of lesion index and image analysis. Moreover, image analysis has been applied to various biochemical data and is one of methods for analyzing amplified PCR bands or protein spots detected by antibodies after electrophoresis and quantitatively calculating the band intensities; it also allows easy counting of cells (or nuclei) with the mammalian cell culture feature,. Furthermore, because it is difficult to quantify immunohistochemical data by observation through naked eyes, image analysis is useful to quantitatively calculate the proportion of stained cells as well as stained areas. The advantages of the image analysis method were described in a previous study in which the results of image analysis were compared by employing the spectrophotometric method. When it was plotted as a graph based on a linear equation, the coefficient of determination (R-square) value indicated data validation.
In conclusion, the current experimental study provided evidence for the protective effects of O. humifusa aqueous extract in a mouse model with ethanol-induced gastric ulcers, which were mediated via suppression of gastric inflammation and oxidative stress. Additionally, this study also reported a high antioxidant activity of this plant extract. HPLC analysis showed a large number of peaks which validated the presence of active compounds. Although chemical drugs for gastric ulcers are commercially available at present, the use of natural materials is preferred. The abovementioned finding highlights the use of O. humifusa aqueous extract as an effective and secure strategy for regulating and managing gastric ulcers. Further information is warranted by exploring the probable potency of O. humifusa aqueous extract as an adjunct for managing gastric ulcers. Moreover, if certain obstacles pertaining to image analysis are overcome, it can be a long-lasting and useful tool for establishing phenotypic properties even for other target diseases.
Conflict of interest statement
The authors declare no conflicts of interest.
Authors’ contributions CYY executed overall on manuscript and experiment. HUS performed analytic calculation of figure data. AF, HJC and MBA contributed to in vivo experiment. SHL supervised this project as a corresponding author.
| References|| |
Li QJ, Wang ZG, Xie Y, Liu Q, Gao YX. Mechanistic evaluation of gastro-protective effects of KangFuXinYe on indomethacin-induced gastric damage in rats. Chin J Nat Med
Hiraishi H, Shimada T, Ivey KJ, Terano A. Role of antioxidant defenses against ethanol-induced damage in cultured rat gastric epithelial cells. J Pharmacol Exp Ther
Ozbakiş Dengiz G, Gürsan N. Effects of Momordica charantia
L. (Cucurbitaceae) on indomethacin-induced ulcer model in rats. Turk J Gastroenterol
Kim YS, Park HJ, Kim H, Song J, Lee D. Gastroprotective effects of Paeonia extract
mixture HT074 against experimental gastric ulcers in rats. Evid Based Complement Alternat Med
Khoder G, Al-Menhali AA, Al-Yassir F, Karam SM. Potential role of probiotics in the management of gastric ulcer. Exp Ther Med
Maity P1, Biswas K, Roy S, Banerjee RK, Bandyopadhyay U. Smoking and the pathogenesis of gastroduodenal ulcer-recent mechanistic update. Mol Cell Biochem
Hatazawa R, Ohno R, Tanigami M, Tanaka A, Takeuchi K. Roles of endogenous prostaglandins and cyclooxygenase isozymes in healing of indomethacin-induced small intestinal lesions in rats. J Pharmacol Exp Ther
Shin S, Jeon JH, Park D, Jang JY, Joo SS, Hwang BY, et al. Antiinflammatory effects of an ethanol extract of Angelica gigas
in a Carrageenan-air pouch inflammation model. Exp Anim
Yang EI, Lee CH, Che DN, Jang SI, Kim YS. Biological activities of water soluble polysaccharides from Opuntia humifusa
stem in high-fat-diet-fed mice. J Food Biochem
Kim HM. Antioxidant activity and skin safety evaluation effects of cactus cladodes (Opuntia humifusa) extract. Master Dissertation. Graduate School of Korea University; 2018
Hahm SW, Park J, Son YS. Opuntia humifusa
stems lower blood glucose and cholesterol levels in streptozotocin-induced diabetic rats. Nutr Res
Lee SY, Bae CS, Choi YH, Seo NS, Na CS, Yoo JC, et al. Opuntia humifusa
modulates morphological changes characteristic of asthma via
IL-4 and IL- 13 in an asthma murine model. Food Nutr Res
Choi SB, Bae GS, Park KC, Jo IJ, Seo SH, Song K, et al. Opuntia humifusa
ameliorated cerulein-induced acute pancreatitis. Pancreas
Han SH, Park K, Kim EY, Ahn SH, Lee HS, Suh HJ. Cactus (Opuntia humifusa)
water extract ameliorates loperamide-induced constipation in rats. BMC Complement Altern Med
Satoh H, Takeuchi K. Management of NSAID/aspirin-induced small intestinal damage by GI-sparing NSAIDs, anti-ulcer drugs and food constituents. Curr Med Chem
Fahmy NM, Al-Sayed E, Michel HE, El-Shazly M, Singab ANB. Gastroprotective effects of Erythrina speciosa
(Fabaceae) leaves cultivated in Egypt against ethanol-induced gastric ulcer in rats. J Ethnopharmacol
Xu P, Yang L, Yuan RY, Ye ZY, Ye HR, Ye M. Structure and preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum
sp. Int J Biol Macromol
Seo SH, Kim JH, Kim JW, Kye YC, Ahn HH. Better understanding of digital photography for skin color measurement: With a special emphasis on light characteristics. Skin Res Technol
Meznaric M, Carni A. Characterisation of flexor digitorum profundus, flexor digitorum superficialis and extensor digitorum communis by electrophoresis and immunohistochemical analysis of myosin heavy chain isoforms in older men. Ann Anat
Horne R, Weinman. Patients’ beliefs about prescribed medicines and their role in adherence to treatment in chronic physical illness. J Psychosom Res
Bhesh Raj Sharma, Park CM, Choi JW, Rhyu DY. Anti-nociceptive and anti-inflammatory effects of the methanolic extract of Opuntia humifusa
stem. Avicenna J Phytomed
Yoon JA, Hahm SW, Park JE, Son YS. Total polyphenol and flavonoid of fruit extract of Opuntia humifusa
and its inhibitory effect on the growth of MCF-7 human breast cancer cells. J Korean
Soc Food Sci Nutr
Kim YJ, Park CI, KIM SJ, Ahn EM. Antioxidant and inflammatory mediators regulation effects of the roots of Opuntia humifusa. J Appl Biol Chem
Noguera ME, Jakoncic J, Ermácora MR. High-resolution structure of intramolecularly proteolyzed human mucin-1 SEA domain. Biochim Biophys Acta Proteins Proteom
Lee C. Gastric mucosa protective activity of Aloe vera milk on ethanol-induced mouse model
. Master Dissertation. Kyungpook National University; 2013.
Ma L, Liu J. The protective activity of Conyza blinii
saponin against acute gastric ulcer induced by ethanol. J Ethnopharmacol
Almasaudi SB, El-Shitany NA, Abbas AT, Abdel-dayem UA, Ali SS, Al Jaouni SK, et al. Antioxidant, anti-inflammatory, and antiulcer potential of Manuka Honey against gastric ulcer in rats. Oxid Med Cell Longev
Arab HH, Salama SA, Omar HA, Arafa el-SA, Maghrabi IA. Diosmin protects against ethanol-induced gastric injury in rats: Novel anti-ulceractions. PLoS One
Subhash VV, Ho B. Inflammation and proliferation - a causal event of host response to Helicobacter pylori
Peng YC, Huang LR, Shyu CL, Cheng CC, Ho SP. Interaction of omeprazole and Helicobacter
pylori-induced nuclear factor-kappaB activation and mediators in gastric epithelial cells. J Chin Med Assoc
Mabrok HB, Mohamed MS. Induction of COX-1, suppression of COX-2 and pro-inflammatory cytokines gene expression by moringa leaves and its aqueous extract in aspirin-induced gastric ulcer rats. Mol Biol Rep
Aziz RS, Siddiqua A, Shahzad M, Shabbir A, Naseem N. Oxyresveratrol ameliorates ethanol-induced gastric ulcer via
downregulation of IL-6, TNF-α, NF-κB, and COX-2 levels, and upregulation of TFF-2 levels. Biomed Pharmacother
Elizalde-Velázquez A, Subbiah S, Anderson TA, Green MJ, Zhao X, Cañas-Carrell JE. Sorption of three common nonsteroidal anti-inflammatory drugs (NSAIDs) to microplastics. Sci Total Environ
Abdul Aziz, Karolin Kamel. Comparative evaluation of the anti-ulcer activity of curcumin and omeprazole during the acute phase of gastric ulcer. Food Nut Sci
Furukawa K, Takahashi T, Arai F, Matsushima K, Asakura H. Enhanced mucosal expression of interleukin-6 mRNA but not of interleukin-8 mRNA at the margin of gastric ulcer in Helicobacter pylori
-positive gastritis. J Gastroenterol
Subedi L, Lee SE, Madiha S, Gaire BP, Jin M, Yumnam S, et al. Phytochemicals against TNFa-mediated neuroinflammatory diseases. Int J Mol Sci
Prasad K, Prabhu GK. Image analysis tools for evaluation of microscopic views of immunohistochemically stained specimen in medical research-a review. J Med Syst
Huleihel L, BenYA, Milosevic J, Yu G, Pandit K, Sakamoto K, et al. Let-7d microRNA affects mesenchymal phenotypic properties of lung fibroblasts. Am J Physiol Lung Cell Mol Physiol
Rhim JH, Luo X, Xu X, Gao D, Zhou T, Li F, et al. A high-content screen identifies compounds promoting the neuronal differentiation and the midbrain dopamine neuron specification of human neural progenitor cells. Sci Rep
Ghali W, Vaudry D, Jouenne T, Marzouki MN. Lycium europaeum
fruit extract: Antiproliferative activity on A549 human lung carcinoma cells and PC12 rat adrenal medulla cancer cells and assessment of its cytotoxicity on cerebellum granule cells. Nutr Cancer
Owens P, Pick MW, Novitskiy SV, Chytil A, Gorska AE, Aakre ME, et al. Disruption of bone morphogenetic protein receptor 2 (BMPR2) in mammary tumors promotes metastases through cell autonomous and paracrine mediators. Proc Natl Acad Sci U S A
Rizzardi AE, Johnson AT, Vogel RI, Pambuccian SE, Henriksen J, Skubitz AP, et al. Quantitative comparison of immunohistochemical staining measured by digital image analysis versus pathologist visual scoring. Diagn Pathol
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6]