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Wound Healing Activity of Kaempferia Rotunda Linn Leaf Extract

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Wound Healing Activity of Kaempferia Rotunda Linn Leaf Extract Int.J.Curr.Microbiol.App.Sci (2013) 2(12): 74-78 ISSN: 2319-7706 Volume 2 Number 12 (2013) pp. 74-78 http://www.ijcmas.com Original Research Article Wound healing activity of Kaempferia rotunda Linn leaf extract Syed Aman Imam, Saroj Kumar Rout, Niranjan Sutar*, Uma Shankar Sharma and Ranju Sutar PG Department of Pharmacognosy, Sir Madanlal Institute of Pharmacy, Etawah -206001, India *Corresponding author A B S T R A C T K e y w o r d s Kaempferia rotunda Linn is an Indian Ayurvedic plant. The present study reports that investigations were done on the aqueous and methanolic extract of Kaempferia Acute toxicity; rotunda Linn leaf by using the incision wounds and excision wounds modals study Excision in rat. At the end of the day the study revealed that the aqueous extract and wounds; methanolic extract displayed significant wound healing activity and justifying the Incision traditional claim. wounds. Introduction The wound may be defined as a loss or Conversely, chronic wounds have breaking of cellular and anatomic or proceeded through portions of the repair functional continuity of living tissues. process without establishing a functional Healing of wound is a biological process anatomic result (Cohen et al., 1999; Joan that is initiated by trauma and often et al., 2003). terminated by scar formation. The process of wound healing occurs in different Many medicinal plants are used in the phases such as coagulation, epithelization, several ways like food, medicine and of granulation, collegenation and tissue other economical and social importance. remodeling. In India, there has been The plant Kaempferia rotunda Linn interest in the potential of medicinal plant belongs to the family Zingiberaceae also for development of drugs with wound named bhuichampaka (Sanskrit), healing properties as taught in a popular bhuchampa (Hindi), and blackhorm form of Indian medicine known as (English). It is a fragment aromatic herb Ayurveda (Jain et al., 2006). with a tuberous rhizome distributed throughout India .In some districts of Two broad categories exist for the Maharashtra the powder root is popular in classification of wounds: chronic and mumps and also said to be used in the acute. Acute wounds undergo a complex form of poultice, promotes suppuration. interactive process involving a variety of The main constituent crotepoxide is useful cell types that leads to a healed wound. 74 Int.J.Curr.Microbiol.App.Sci (2013) 2(12): 74-78 for the inhibition of tumors (Kupchun et 25 -30 gm were employed for this study al., 1969). Phytochemical the plant has and were procured from institute animal been attributed to contain flavonoids, house. They were fed with standard diet crotepoxide, chalcones, quercetin, and water and housed in cages at room flavonols, ß sitosterols, stigmasterol, temperature (30±2°C) with a 12 h light syringic acid, protocatechuic acid and and dark cycle. Ethical clearance required some hydrocarbons have been previously for the animal experiment was obtained reported (Pai et al., 1970). from institute animal ethical committee (IAEC). Materials and Methods Acute toxicity studies (LD50) Plant material Acute toxicity study of aqueous and The leaves of Kaempferia rotunda Linn methanolic extract of Kaempferia rotunda were collected from local market of was performed in albino rats according to etawah (Uttar Pradesh). The plant was OECD guidelines 423 (OECD, 1996). The identified at Pharmacognosy department animals were kept fasting for overnight of Sir Madanlal Institute of Pharmacy, providing only water, after administration Etawah (UP), and India. The voucher aqueous and methanolic extract orally at specimen of Kaempferia rotunda Linn (kr- doses of 5 2000 mg/kg. Animals were 14) has been preserved in our herbarium then allowed free access to food and water for further collection and reference. and observed a period of 48 hrs for signs of acute toxicity. The number of deaths Preparation of extracts within this period was recorded. The leaves were washed thoroughly, dried Wound models under a shade and pulverized. The coarse powder was extracted with aqueous and The studies were carried out using 95% methanol using a soxhlet apparatus. ether anesthetized rats and their back was The Kaempferia rotunda extracts were shaved. The selected animals were divided dried using a rotary vacuum evaporator into two groups of six in each. All the test and stored in a desiccators until further samples were applied topically models at use. two different dose levels of 250 and 500 mg/kg body weight. Phytochemical screening Incision wounds The aqueous and methanolic extract of Kaempferia rotunda were subjected to Two Para vertebral incisions (6 cm long) phytochemical screening (Kokate, 1994; were made through the full thickness of Khandelwal, 2004) to identify the nature the skin on either side of the vertebral of chemical constituents present in the column of the rat (Lee, 1968). Wounds plant material. were closed with interrupted sutures, 1 cm apart. The sutures were removed on the Animals seventh day. Wound breaking strength was measured in anesthetized rats on the Wister albino rats of weighing about 150- tenth day after wounding (Morton and 200 gm and adult albino mice of weighing Malone, 1972). 75 Int.J.Curr.Microbiol.App.Sci (2013) 2(12): 74-78 Table.1 Incision modal Solvents Breaking strength(g) Control 211.333±4.780 aqueous extract of Kaempferia rotunda 250mg/kg 367.105±2.140 aqueous extract of Kaempferia rotunda 500mg/kg 327.653±1.865 Methanolic extract of Kaempferia rotunda 415.983±1.654 250mg/kg methanolic extract of Kaempferia rotunda 390.653±1.974 500mg/kg Table.2 Excision modal Wound model Excision % of wound contraction Parameter 4th 8th 12th Studies Control 23.577 ±0.165 33.473±0.079 53.667 ±0.123 aqueous extract of Kaempferia 25.373 ±0.164 46.587 ±0.096 80.387 ±0.033 rotunda 250mg/kg aqueous extract of Kaempferia 26.480 ±0.320 52.796±0.094 83.594±0.077 rotunda500mg/kg methanolic extract of Kaempferia 26.323 ±0.163 49.537 ±0.099 87.347,±0.075 rotunda250mg/kg methanolic extract of Kaempferia 26.870 ±0.320 59.757±0.077 93.503 ±0.074 rotunda500mg/kg Figure.1 Break strength 76 Int.J.Curr.Microbiol.App.Sci (2013) 2(12): 74-78 Figure.2 Excision % of wound contraction Excision wounds Acute toxicity test (LD50) A circular skin piece of full thickness Oral administration of graded doses (5 to (approximately 500 mm²) was removed 2000 mg/kg ) of the aqueous and from a predetermined dorsal area (Neuman methanolic extract of Kaempferia rotunda and Logan, 1950). The wounds were to rats did not produce any significant traced on 1 mm² graph paper on the day of changes in behavior, breathing, cutaneous wounding and subsequently on alternate effects, sensory nervous system responses days until healing were complete. Changes or gastrointestinal effects during the in the wound area were calculated, giving observation period. No mortality was an indication of the rate of wound recorded in any group after 72 hr of contraction. administering the extract to the animals. Statistical analysis Wound healing All the results are expressed as mean ± The results of plant in wound healing standard error. The data was analyzed property are given in the table-1 and table- statistically using ANOVA. 2. Result and Discussion Wound healing process consists of different phases such as granulation, Phytochemical screening collagenation, collagen maturation and The phytochemical tests revealed the scar maturation which are concurrent but presence of Flavonoids, crotepoxid, independent to each other. Hence in the chalcones, quercetin, protocatechuic acid, present study two different wound models -sistosterol, stigmosterol, syringic acid, were used. In the incision wound model, a and some hydrocarbons in methanol significant increase was observed in the extract. skin breaking strength of the aqueous and 77 Int.J.Curr.Microbiol.App.Sci (2013) 2(12): 74-78 methanolic extract treated group at both Khandelwal, K.R., 2004. Practical dose levels (Table 1). The study on the Pharmacognosy , 11th ed. Nirali excision wound animal model revealed Prakashan, Pune, 2004. that the drug extract affect the rate of Kokate, C.K., 1994. Practical wound contraction significantly at both Pharmacognosy, 4 th ed, Vallaph dose levels (Table 2). Prakashan, Delhi, 1994. Kupchun, S.M., R.J. Hemingway andSmith RM. Crotepoxide , a novel The above finding provides an insight into cyclohexane diepoxide tumor inhibitor the usage of Kaempferia rotunda Linn leaf from Croton macrostachys. J. Org. in traditional treatment of wounds, burns Chem. 34:3898-902. and reduces swelling. Lee, K.H., 1968. Studies on the mechanism of action of salicylates II. Acknowledgement Effects of Vitamin A on Wound Healing retardation action of aspirin. J. The present study is carried out in the Sir Pharm. Sci. 57:1238. Madanlal group of Intitutions and the Morton, J.J.P, and Malone, H.H. 1972. authors are very much thankful to Evaluation of vulnerary activity by chairman Mr. Vivek Yadav for providing open wound procedure in rats, Arch Int all facilities. Pharmacodyn. 196: 117. Neuman, R.E., and Logan, M.A. 1950. References The determination of collagen and elastin in tissues. J. Biochem. 186: 549 Cohen, I.K., R.F. Diegelman, R.Y. Dome, OECD, 1996. OECD Guidelines for the S,I, Schwartz, G.T. Shires, F.C. testing of chemicals, Test no.423; Spencer et al., 1999. Wound care and Acute Oral Toxicity Acute Toxic wound healing. In: Principles of Class Method, 1996. surgery. seventh edition. New York: Pai, B.R., N.N. Rao and Wariyor, N.S . McGraw-Hill. p. 263 95. 1970. Occurence of crotepoxide in Jain, V., V. Prasad and Pandey, R.S. Kaempferia rotunda linn. Indian. J. 2006. Wound healing activity of Chem. 8:468. Desmodium gangeticum in Different Wound Model.
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