Evaluation of Anticolitis and Antioxidant Properties of Bixa orellana (Bixaceae) Leaf Hydroethanolic Extract on Acetic Acid-Induced Ulcerative Colitis in Rats.

Background: Ulcerative colitis is an idiopathic inflammatory bowel disease characterized by tissue damage, diarrhea, anemia, and loss of body weight. Tissue damage occurs as a result of uncontrolled activation of the immune response and an increase in free radicals, which have a strong effect on the pathogenesis of inflammatory bowel disease. The incidence and prevalence of this inflammatory disease continue to increase worldwide. Maceration of Bixa orellana leaves in palm wine is used in traditional medicine to treat diarrhea, dysentery, and hemorrhoids in the Adamaoua region of Cameroon. Objective: The present work evaluated the preclinical effects (ie, antioxidant, hematological, and histological activities) of the hydroethanolic extract of Bixa orellana leaves in an in vivo, rat acetic acid-induced ulcerative colitis model. Methods: Thirty-six female rats weighing between 165 and 180 g were fasted for 18 hours and then anesthetized with ether. A dose of 1 mL acetic acid (5%) was administered rectally through a catheter in all rats except the normal control group, which received distilled water (1 mL) instead. Treatments began 48 hours after rectal administrations of acetic acid or water, and all animals were treated twice daily for 7 days. The normal control group and the colitis control group received PO distilled water (10 mL/kg), the positive control received orally loperamide (5 mg/kg, and the 3 test groups received orally the hydroethanolic extract of Bixa orellana at 100, 200, and 400 mg/kg, respectively. During treatment, the number of diarrheal stools and weight change were assessed. At the end of the treatment, the animals were put to death under ether anesthesia. Blood was collected postmortem for evaluation of hematological and antioxidant parameters. The abdomen was opened via a midline incision and the colon was removed and emptied of all contents to assess histological and antioxidant parameters. Results: During treatment, the number of diarrheal stools was significantly decreased from day 3 in animals treated with 100 (P < 0.05), 200 (P < 0.05), and 400 (P < 0.01) mg/kg extract compared with the colitis control group. The change in body weight of all extract-treated rats decreased significantly from day 3 (-5.55%; P < 0.05) to day 8 (-13.80%; P < 0.01) compared with the normal control. In the colitis control, this change ranges from -6.15% on day 2 to -15.13% on day 8. Extract treatment with 100, 200, and 400 mg/kg significantly reduced (P < 0.05) the number of lesions and the relative weight of the colon. The levels of red blood cells, neutrophils, and total white blood cells decreased in the colitis control group, whereas treatment with the extract at doses of 100, 200, and 400 mg/kg was associated with a significant increase in these hematological parameters. Catalase and superoxide dismutase activity and glutathione concentrations all increased significantly (P < 0.01) in blood and colon in all extract-treated animals, whereas levels of malondialdehyde and nitric oxide were significantly decreased (P < 0.01) compared with the colitis control animals. Conclusions: The hydroethanolic extract of Bixa orellana leaves had protective effects against acetic acid-induced ulcerative colitis in rats that was associated with inhibited production of free radicals believed to be responsible for oxidative stress, hematological disorders, and tissue damage in this animal model.

Anxiolytic and anti-colitis effects of Moringa oleifera leaf-aqueous extract on acetic acid-induced colon inflammation in rat.

Moringa oleifera decoction is believed to alleviate gastrointestinal tract diseases. This study investigated antioxidant and anxiolytic activities of its leaves aqueous extract on acetic acid-induced colitis in rats. Rats (36) were randomly divided into six groups and received (20 days) distilled water, 10 mL/kg; Moringa oleifera leaf-aqueous extract (25, 50, and 100 mg/kg) or Loperamide (5 mg/kg). On days 1, 8, 17, and 20, behavioral parameters were evaluated. Colitis was induced (day 15, except in normal group) through acetic acid (4%, 1 mL) intra-rectal administration. After sacrifice (day 21), lesion number, weight/length ratio of the colon were recorded. Oxidative stress biomarkers were evaluated. On day 20, Moringa oleifera (100 mg/kg) reduced the number of head dipping and the duration in opened arms, respectively 2.00 ± 0.37 and 5.00 ± 0.37 s against 14.50 ± 0.72 and 2.17 ± 0.48 s in the control. It decreased colon weight/length ratio: 112.29 ± 9.46 against 185.93 ± 5.28 mg/cm in the control; malondialdehyde level (P < 0.01) and nitric oxide concentration (P < 0.001), in the brain: respectively 25.60 ± 0.60 and 36.34 ± 1.19 against 34.00 ± 0.33 and 46.17 ± 3.25 µmol/mg of tissue in the control. In the serum, the extract (50 mg/kg) significantly (P < 0.05) increased the catalase activity (0.10 ± 0.00 against 0.03 ± 0.00 µmol/mg of protein in the negative control group). At 100 mg/kg, it increased (P < 0.001) reduced glutathione concentration to 5.07 ± 0.31 against 3.26 ± 0.08 µmol/mg of protein in the negative control group. The improvement on colitis pathophysiology, the antioxidant and the anxiolytic effects noted therefore suggest that Moringa oleifera can be a potential source of drugs alleviating anxiety and oxidative stress associated to ulcerative colitis.

Anti-Shigellosis Activity of Cola anomala Water/Ethanol Pods Extract on Shigella flexneri-Induced Diarrhea in Rats.

This study was undertaken to evaluate the activities of water/ethanol Cola anomala pods extract. In vitro antimicrobial susceptibility was determined by the disk diffusion method; the minimum inhibitory concentration and minimum bactericidal concentration were determined by agar dilution technique. In vivo, shigellosis was induced in healthy Wistar albino rats by oral administration of Shigella flexneri inoculum, 12 × 108 CFU/mL. At the onset of diarrhea, infected and normal control animals were subdivided into various groups treated with distilled water, with water/ethanol Cola anomala pods extract at 25, 50, or 100 mg/kg, or with ciprofloxacin, 2.5 mg/kg. After one-week treatment, rats were sacrificed, and blood and colon were collected. Blood was used for blood cell count. A portion of the colon served for histological studies while homogenate from the remaining part was centrifuged and the supernatant was collected for the determination of NO, PGE2, IL-1ß, and TNF-α levels. In vitro, water/ethanol Cola anomala pods extract showed to be bactericidal, with a minimum inhibitory concentration of 2.0 mg/mL and a minimum bactericidal concentration of 3.0 mg/mL. In diarrheic rats, the extract significantly (P < 0.01) increased the white blood cells and significantly (P < 0.01) decreased stool Shigella density from the first to the seventh day of treatment. It partially restored the structure of eroded intestine epithelium and prevented weight loss; the dose dependently and significantly (P < 0.001) decreased NO, IL-1ß, and TNF-α production in the colon and was found to have no significant effect on PGE2 production. These results support the use of this plant in traditional medicine in the treatment of gastrointestinal ailments.

Effect of the Hydroethanolic Extract of Bixa orellana Linn (Bixaceae) Leaves on Castor Oil-Induced Diarrhea in Swiss Albino Mice.

OBJECTIVE: The treatment of diarrheal diseases is a serious problem in developing countries, where population generally uses medicinal plants. The leaves of Bixa orellana have been reported to be traditionally used in the treatment of diarrhea by local people in the district of Khulna in Bangladesh. The aim of this study was to investigate the effects of the hydroethanolic extract of Bixa orellana leaves on castor oil-induced diarrhea in mice. METHODS: The powder of the leaves of Bixa orellana was macerated in ethanol/water mixture (20/80) for 48 hours and then filtered. The filtrate obtained was lyophilized, and the solutions to be administered to the animals were prepared. To induce diarrhea, animals orally received castor oil (1 mL/100 g bw). To determine the effective doses, each mouse received, 30 minutes after the administration of castor oil, one of the single oral doses of hydroethanolic extract of Bixa orellana leaves: 0, 25, 50, 100, and 200 mg/kg bw. The mass, number, and frequency of stool diarrhea were measured and recorded per hour for five hours. The effect of the hydroethanolic extract of Bixa orellana leaves on the intestinal transit was evaluated by measuring the distance traveled by the charcoal meal in thirty minutes. The effects of the aqueous extract of hydroethanolic extract of Bixa orellana leaves on intestinal secretion were evaluated by measuring the volume of the intestinal content and by dosing the electrolytes (Na+, K+, and Cl-) in the intestinal content by the colorimetric method. RESULTS: The extract produced significant (P < 0.01) decreases, respectively, 35.52%, 54.47%, 74.80%, and 87.80% in the severity of diarrhea. The extract at 100 and 200 mg/kg bw showed a significant (P < 0.01) decrease of castor oil-induced enteropooling (61.08% and 65.41%), and only the 200 mg/kg bw exhibited significant (P < 0.01) reduction on intestinal transit (24.46%) as compared to standard drug. CONCLUSIONS: The hydroethanolic extract was found to be effective against castor oil-induced diarrhea in experimental mice at 50, 100, and 200 mg/kg bw which provides evidence that could justify its traditional use.

Antioxidant activity of dichloromethane fraction of Dichrocephala integrifolia in Salmonella typhi-infected rats.

OBJECTIVE: Infectious diseases such as typhoid fever lead to the formation of free radicals which can damage the body. Many medicinal plants have antioxidant molecules that neutralize free radicals. The present work evaluated the antioxidant activity and histopathological effects of the dichloromethane fraction of Dichrocephala integrifolia in Salmonella typhi-infected rats. METHODS: The S. typhi-infected rats concurrently received daily doses of D. integrifolia extract at doses of 25, 50 and 100 mg/kg body weight or ciprofloxacin (5 mg/kg body weight) for 15 days. Body temperature was measured daily during infection and treatment periods. At the end of treatment period, the animals were sacrificed and biological responses including hematological parameters, superoxide dismutase and catalase activities, and glutathione, malondialdehyde and nitric oxide concentrations were evaluated. RESULTS: The elevated body temperature induced by infection was significantly decreased in animals treated with 25, 50 or 100 mg/kg of the extract. Platelet levels decreased slightly in infected rats, while treatment with the dichloromethane fraction of D. integrifolia significantly increased platelet levels; this response was greater than that elicited by ciprofloxacin. The doses of 50 and 100 mg/kg of the dichloromethane fraction of D. integrifolia notably decreased monocyte and neutrophil values. Activity of superoxide dismutase and catalase and levels of glutathione in the tissues of treated animals were increased significantly (P < 0.01), while malondialdehyde and nitric oxide levels were significantly decreased (P < 0.01), following treatment with the dichloromethane fraction of D. integrifolia. CONCLUSION: The results of this study show that the dichloromethane fraction of D. integrifolia has protective effects against a series of pathological conditions initiated by oxidation and tissue damage in the course of a S. typhi infection.