Pharmacological investigations

1. Anti-inflammatory activity: The objective of the present study was to investigate the anti-inflammatory property of the aqueous extract of the leaves of H. integrifolia. The hind-paw edema was produced in rats by the sub-planter injection of carageenan. The aqueous extract of H. integrifolia at dose (250 and 500 mg/kg p+.o) was given to observe % inhibition of paw edema which were comparable with indomethacin

(10 mg/kg p.o) used as a reference drug. The extract administered orally at doses of 250 and 500 mg/kg p.o produced a significant (P < 0.05) dose dependent inhibition of edema formation.

2. Anti-nausea activity: In a rat model, anti-emetic activity of ethanolic extract of leaves of H. integrifolia on cisplatin-induced nausea was investigated. Rats react to emetic/nausea-producing stimuli, such as cisplatin, with altered feeding habits, manifested by pica or increased consumption of kaolin. The authors measured pica in rats to quantify cisplatin-induced nausea, and to evaluate the anti nausea effect of pretreatment with ethanolic extract of H. integrifolia, which was given orally. Cisplatin at 3 mg/kg (i.p) induced significant pica accompanied by reduced food intake, suggesting the presence of nausea. Hence, this cisplatin dose was selected for testing the anti nausea activity of H. integrifolia. Cisplatin-induced pica decreased significantly when animals were pretreated with H. integrifolia at doses of 250 mg/kg p.o and 500 mg/kg p.o (P<0.01). H. integrifolia pretreatment decreased cisplatin induced kaolin intake in the rat model of simulated nausea. The findings suggest that H. integrifolia and its active constituent(s) may offer a therapeutic role in chemotherapy-induced emesis.

3. Antimicrobial, antioxidant and wound-healing activity: A. The methanolic extracts of H. integrifolia leaves and stem bark were studied for its wound-healing potential. Since wound healing is severely hampered by microbial infection and reactive oxygen species (ROS), this study was undertaken to evaluate antimicrobial and antioxidant activity apart from wound-healing activity. The antimicrobial property of the H. integrifolia was studied against six bacterial and five fungal strains using the agar well diffusion method and minimum microbicidal concentration and minimum inhibitory concentration were determined for each strain. Methanolic extract of stem bark showed a larger zone of inhibition (11.3-20.4 mm) than methanolic extract of leaves (9.6-14.9 mm).

The anti-oxidant activity was evaluated by DPPH free radical scavenging activity using HPLC method. The IC50 values obtained for methanolic extract of stem bark (TPC: 78.53 ± 1.26 mg/g) and methanolic extract of leaves (TPC: 57.71 ± 1.45 mg/g) were 37.66 ± 0.48 and 50.36 ± 0.59 ^g/well, respectively. In excision wound model, more than 90% wound healing was recorded in treated groups by 14 d of post surgery, where as only 62.99% was observed in the control group. In incision model, higher breaking strengths and higher hydroxyproline content in treated groups suggested higher collagen re-deposition than the control group. Further, histopathology studies conformed wound-healing activity of H. integrifolia.

B. In another study, the ethanolic crude extract of stem bark of H. integrifolia was screened for its antioxidant activity using a-tocopherol as standard antioxidant. The free radical scavenging potential of the extract was evaluated by two different antioxidant methods; ferric thiocyanate and thiobarbituric acid method. The ethanol extract was found to exhibit good antioxidant property.

C. The inhibitory effect of a phytochemical, 1, 4-naphthalenedione, isolated from H. integrifolia on p-lactamase is reported here. This compound was found to have a synergistic effect with the antibiotic amoxicillin against a resistant strain of Staphylococcus aureus. The enzyme was purified from the organism and incubated with the compound. An assay showed that the compound can inhibit the enzymatic activity of p-lactamase. Modeling and molecular docking studies indicated that the compound can fit into the active site of a-lactamase. Hence, the compound can serve as a potential lead compound for the development of effective p-lactamase inhibitor that can be used against p-lactam-resistant microbial strains.

4. Antidiarrheal activity: The ethanolic extract of leaves of H. integrifolia was studied for its antidiarrheal properties in experimental diarrhea, induced by castor oil and magnesium sulfate in mice. At the doses of 250 and 500 mg/kg per oral, the ethanolic extract showed significant and dose-dependent antidiarrheal activity in both models. The extracts also significantly reduced the intestinal transit in charcoal meal test when compared to atropine sulate (5 mg/kg; i.m.). The results showed that the ethanolic extract of leaves of H. integrifolia have a significant antidiarrheal activity and supports its traditional uses in herbal medicine.

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