{"id":259,"date":"2015-01-20T07:50:08","date_gmt":"2015-01-20T07:50:08","guid":{"rendered":"http:\/\/biomedpharmajournal.org\/?p=259"},"modified":"2017-01-04T07:41:11","modified_gmt":"2017-01-04T07:41:11","slug":"phytochemical-analysis-and-anti-inflammatory-activity-of-pisonia-grandis-r-br","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol1no1\/phytochemical-analysis-and-anti-inflammatory-activity-of-pisonia-grandis-r-br\/","title":{"rendered":"Phytochemical Analysis and Anti-Inflammatory Activity of Pisonia Grandis R.Br."},"content":{"rendered":"

Introduction<\/strong><\/p>\n

The World Health Organization (WHO) estimated that 80% of the population of developing countries still relies on traditional medicines, mostly plant drugs, for their primary health care (Fransworth et al., 1985).\u00a0 Plant products also play an important role in the healthcare systems of the remaining 20% population, mainly residents of developed countries.\u00a0 The scientific data generated by research on the plants serves as a valuable tool for identifying plant species and for characterization of the pharmacological active constituent for their biological activities.\u00a0 In the search for new plant it is always important to screen for its activity as first step.\u00a0 Once the plant is identified for beneficial biological activity it is imperative to collect supporting scientific\u00a0\u00a0 data generated through pharmacognostic and phytochemical properties of plant under investigation.<\/p>\n

Based on this we have worked on the leaves of Pisonia grandis<\/em> R.Br. (Nyctaginaceae) to find out anti-inflammatory activity. P. grandis<\/em> is an herb claimed to be used for treatment of wound healing, algesia, ulcer and antibacterial activity (Kirithikar and Basu 1990; Prabu et al., 2008).\u00a0 Alcohol and aqueous extracts were taken and analyzed for anti-inflammatory activity.\u00a0 This study was aimed giving vital information regarding pharmacological and phytochemical activities.\u00a0 As a positive step is taken in this direction for achieving betterment of mankind.\u00a0 A small step has been taken through this research.<\/p>\n

Materials and Methods<\/strong><\/p>\n

Plant collection<\/strong><\/p>\n

The leaves of Pisonia grandis<\/em> were collected in Chengalpattu (Tamil Nadu) in June 2006 and authenticated by a botanist Dr. P. Jayaraman at the Plant Anatomy Research Centre (PARC), Chennai, Tamil Nadu, India.\u00a0 A voucher specimen has been deposited at the museum of the department of pharmacognosy, Madras Medical College, Chennai, India.<\/p>\n

Preparation of extracts<\/strong><\/p>\n

The freshly collected leaves of Pisonia grandis<\/em> were shade dried and then coarsely powdered in a blender.\u00a0 The coarse powder was successively extracted in an aspirated bottle with ethanol, water by cold maceration for 3-7 days.\u00a0 After decantation and filtering through What Mann filter paper no.41 nearly 81% of the solvent was removed by distillation over boiling water bath and remaining under reduced pressure.\u00a0 The extracts so obtained were further dried in vacuum desiccators and the residue obtained from various extracts was used for further studies by preserving it in refrigerator.<\/p>\n

Phytochemical studies<\/strong><\/p>\n

The presence of phytochemicals, triterpenoids (Noller\u2019s test), flavones (Shinadow\u2019s test), steroids (Libermann-Burchard test), proteins (Biuret test), furans (Ehrlich\u2019s test), alkaloids (Dragendroff\u2019s reagent), anthraquinones (Borntrager\u2019s test), gums, tannins (5% ferric chloride), saponins (Frothing test), phenols and sugars were evaluated according to the method described by Edeogal et al., 2005.<\/p>\n

Animals<\/strong><\/p>\n

Inbred Male and female Wistar albino rats (160-200 g) were procured from the animal experimental laboratory of Madras Medical College and used throughout the study.\u00a0 The study was conducted after obtaining Institutional animal ethical committee\u2019s clearance (20\/236\/Aug\u2019 2006).\u00a0 The animals were maintained in colony cages at 25\u00b12o<\/sup>C, relative humidity of 45-55% maintained under 12 h light and dark cycles.\u00a0 The animals were fed with standard animals feed (Hindustan Lever Ltd.) and water adlibitum.\u00a0 All the animals were acclimatized for a week before use and they were maintained in hygienic environment in our animal house.<\/p>\n

Table 1:\u00a0Phytochemical constitutes of Pisonia grandis<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Chemical constituents<\/strong><\/td>\nAlcohol extract<\/strong><\/td>\nAqueous extract<\/strong><\/td>\n<\/tr>\n
Terpenoids<\/td>\n+<\/td>\n–<\/td>\n<\/tr>\n
Flavones<\/td>\n+<\/td>\n–<\/td>\n<\/tr>\n
Steroids<\/td>\n+<\/td>\n–<\/td>\n<\/tr>\n
Proteins<\/td>\n–<\/td>\n–<\/td>\n<\/tr>\n
Furan<\/td>\n+<\/td>\n–<\/td>\n<\/tr>\n
Alkaloids<\/td>\n–<\/td>\n–<\/td>\n<\/tr>\n
Anthraquinone<\/td>\n+<\/td>\n–<\/td>\n<\/tr>\n
Gum<\/td>\n–<\/td>\n–<\/td>\n<\/tr>\n
Tannins<\/td>\n+<\/td>\n+<\/td>\n<\/tr>\n
Saponins<\/td>\n+<\/td>\n–<\/td>\n<\/tr>\n
Phenols<\/td>\n–<\/td>\n–<\/td>\n<\/tr>\n
Sugars<\/td>\n–<\/td>\n–<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Acute toxicity study (Ecobian, 1997)<\/strong><\/p>\n

The procedure was followed by using OECD (Organization of Economic Cooperation and Development) guidelines 423 (Acute toxic class method).\u00a0 Male wistar rats weighing 160-200 gm were used for the study.\u00a0 The starting dose level of poly herbal formulation was 2000 mg\/kg body weight p.o.\u00a0 Dose volume was administered 0.1 ml\/10 gm body weight to the rate which was fated overnight with water adlibitum.\u00a0 Food was with held for the further three to four hours after administration of the drug.\u00a0 Body weight of the rats before and after termination were noted and any changes in skin and fur, eyes and mucous membrane and also respiratory, circulatory, autonomic and central nervous system and locomotors activity and behavior pattern were observed and also sign of tremors, conversion, salivations, diarrhea, lethargy, sleep and coma were noted.\u00a0 The onset of toxicity and signs of toxicity are also noted.<\/p>\n

Anti-inflammatory evaluation<\/strong><\/p>\n

The following experimental protocol was used to access anti-inflammatory activity on carrageenan induced paw edema rats (Wister et al., 1962).\u00a0 The animals were divided into six groups.\u00a0 Each group composed of six animals.<\/p>\n\n\n\n\n\n\n\n\n\n
Group<\/strong><\/td>\nAnimals<\/strong><\/td>\n<\/tr>\n
I<\/td>\nAnimals received 1% carboxy methylcellulose 10 ml\/kg p.o<\/td>\n<\/tr>\n
II<\/td>\nAnimals received aqueous extract of Pisonia grandis<\/em> 200 mg\/kg p.o<\/td>\n<\/tr>\n
III<\/td>\nAnimals received aqueous extract of Pisonia grandis<\/em> 400 mg\/kg p.o<\/td>\n<\/tr>\n
IV<\/td>\nAnimals received alcoholic extract of Pisonia grandis<\/em> 200 mg\/kg p.o<\/td>\n<\/tr>\n
V<\/td>\nAnimals received alcoholic extract of Pisonia grandis<\/em> 400 mg\/kg p.o<\/td>\n<\/tr>\n
VI<\/td>\nAnimals administered reference standard diclofenac sodium 5 mg\/kg p.o<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Paw edema was induced by injecting 0.1 ml of carrageenan in physiological saline into sub-plantar tissue of rat right hind paw of each rat.\u00a0 The aqueous and alcoholic leaf extract of Pisonia grandis<\/em> was administrated orally 30 minutes prior to carrageenan administration.\u00a0 The paw volume was measured at intervals of 1st<\/sup> hour, 2nd<\/sup> hour, 3rd<\/sup> hour and 4th<\/sup> hour by the mercury displacement method using plethysymograph.\u00a0 The percentage inhibition of paw volume in drug treated group was compared with the carrageenan control group (Group I).\u00a0 The diclofenac sodium was used as a reference standard.<\/p>\n

Statistical analysis<\/strong><\/p>\n

All treated groups were compared with the control group (Group I) and the results were analyzed statistically using ANOVA and followed by Dunnet\u2019s test to identify the difference between treated groups and control.\u00a0 The data were considered significant at\u00a0 \u00a0p <0.05.<\/p>\n

Results and discussion<\/strong><\/p>\n

Phytochemical analysis<\/strong><\/p>\n

The result of phytochemicals is listed in the table 1.<\/p>\n

Anti-inflammatory activity<\/strong><\/p>\n

Carrageenan induced paw edema is the most widely used method to screen anti-inflammatory agents.\u00a0 The development of carrageenan induced edema is biphasic, the initial phase is attributed to release of histamine, 5-hydrozytryptamine and kinins in the first hour after injection of carrageenan and the most pronounced second phase is related to release of prostaglandin like substances in 2-3 hours.<\/p>\n

The aqueous and alcoholic extract of Pisonia grandis<\/em> shows significant anti-inflammatory activity against carrageenan injection (Table 2), suggesting that it predominantly inhibits the release of inflammatory mediators form phylogenic stimuli.\u00a0 However, further studies are necessary to identify and isolate the active constituents responsible for its anti-inflammatory activity and also there is a need to elucidate its mechanism of anti-inflammatory action.\u00a0 The present study clearly demonstrated the anti-inflammatory property of Pisonia grandis<\/em>.\u00a0\u00a0 The effects may due to flavonoids, steroids and tannins observed in the extracts.\u00a0 A detailed investigation into the potential plant constituents responsible for the anti-inflammatory property may provide scope for lead molecules that may be useful in treating humans effectively for inflammation.\u00a0 The acute toxicity study did not show any mortality at the dose level of 2000 mg\/kg body weight, so that the extract is characterized as \u201cX\u201d unclassified (OECD-423), this indicates the safety profile of the extracts.\u00a0 The anti-inflammatory activity of Pisonia grandis<\/em> leaf extracts exhibited at the dose level of 200, 400 mg\/kg p.o. exhibited significant (P<0.05) activity compared with the carrageenan treated animals.\u00a0 The reference standard diclofenac sodium also exhibited significant activity.<\/p>\n

Table 2:\u00a0 Anti-inflammatory activity of alcohol and aqueous extract of P. grandis<\/em> on carrageenan induced paw edema rats.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Groups<\/strong><\/td>\n1st<\/sup> hour<\/strong><\/td>\n2nd<\/sup> hour<\/strong><\/td>\n3rd<\/sup> hour<\/strong><\/td>\n4th<\/sup> hour<\/strong><\/td>\n<\/tr>\n
I<\/td>\n0.70\u00b10.16<\/td>\n0.67\u00b10.19<\/td>\n0.65\u00b10.11<\/td>\n0.58\u00b10.06<\/td>\n<\/tr>\n
II<\/td>\n0.58\u00b10.08*<\/td>\n0.52\u00b10.03*<\/td>\n0.51\u00b10.16*<\/td>\n0.44\u00b10.05**<\/td>\n<\/tr>\n
<\/td>\n(17.14%)<\/td>\n(22.38%)<\/td>\n(21.53%)<\/td>\n(24.13%)<\/td>\n<\/tr>\n
III<\/td>\n0.50\u00b10.06**<\/td>\n0.34\u00b10.06**<\/td>\n0.38\u00b10.09**<\/td>\n0.27\u00b10.06**<\/td>\n<\/tr>\n
<\/td>\n(28.57%)<\/td>\n(49.25%)<\/td>\n(41.53%)<\/td>\n(53.47%)<\/td>\n<\/tr>\n
IV<\/td>\n0.39\u00b10.06**<\/td>\n0.37\u00b10.05**<\/td>\n0.36\u00b10.07**<\/td>\n0.33\u00b10.05**<\/td>\n<\/tr>\n
<\/td>\n(44.28%)<\/td>\n(44.77%)<\/td>\n(44.61%)<\/td>\n(43.1%)<\/td>\n<\/tr>\n
V<\/td>\n0.38\u00b10.05**<\/td>\n0.36\u00b10.09**<\/td>\n0.35\u00b10.05**<\/td>\n0.19\u00b10.03**<\/td>\n<\/tr>\n
<\/td>\n(45.71%)<\/td>\n(46.26%)<\/td>\n(46.15%)<\/td>\n(67.24%)<\/td>\n<\/tr>\n
VI<\/td>\n0.16\u00b10.01**<\/td>\n0.14\u00b10.02**<\/td>\n0.16\u00b10.01**<\/td>\n0.13\u00b10.01**<\/td>\n<\/tr>\n
<\/td>\n(77.14%)<\/td>\n(76.11%)<\/td>\n(75.38%)<\/td>\n(77.58%)<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Values are mean \u00b1 S.D.\u00a0 Paw volume expressed in ml, comparison were made between Group I Vs Group II, III, IV, V and VI.\u00a0 Percentage protection given in parenthesis.<\/p>\n

* p<0.05, **p<0.001 statistical evaluation done by ANOVA followed by dunnet\u2019s test.<\/p>\n

Conclusion<\/strong><\/p>\n

The present study indicates that the plant contains potential anti-inflammatory components such as flavonoids, terpenoids and steroids that may be of use for development of phytomedicine for the therapy of inflammations.\u00a0 Further research work is to needed to establish the exact anti-inflammatory mechanism of action of alcoholic extract of Pisonia grandis<\/em>.<\/p>\n

Reference<\/strong><\/p>\n

    \n
  1. Ecobian, D.J., 1997.\u00a0 The basis of toxicity testing, 2nd<\/sup> Ed, CRC press, Newyork, 43-88.<\/li>\n
  2. Edeogal, H.O., Okwu, D.E., M baebie B.O., 2005.\u00a0 Phytochemical constituents of some Nigerian medicinal plants.\u00a0 African Journal of Biotechnology 4, 685-688.<\/li>\n
  3. Fransworth, N.R., Akerele, O., Bingel, A.S., Soejarto, D.D., Guo, Z., 1985.\u00a0 Medicinal plants in therapy. Bull WHO 63, 965-981.<\/li>\n
  4. Kirithikar, K.R., and Basu, B.D., 1990. Indian Medicinal plants. Publication and Information Division, CSIR, New Delhi, India, 2048-2050.<\/li>\n
  5. Prabu, P., Nappinnai, M., Ponnudurai, K., Prabhu, K., 2008.\u00a0 Evaluation of wound healing potential of Pisonia grandis R.Br.:\u00a0 A Preclinical study in wistar rats.\u00a0 The International Journal of Lower Extremity Wounds 7(1): 21-27.<\/li>\n
  6. Wister, C.A., Risley, E.A., and Nuss, G.W., 1962.\u00a0 Carrageenan induced in hind paw of the rats as an assay for anti-inflammatory drugs. Proc. Soc.<\/li>\n<\/ol>\n

     <\/p>\n","protected":false},"excerpt":{"rendered":"

    Introduction The World Health Organization (WHO) estimated that 80% of […]<\/p>\n","protected":false},"author":3,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[4],"tags":[],"class_list":["post-259","post","type-post","status-publish","format-standard","hentry","category-vol1no1"],"_links":{"self":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/259","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/comments?post=259"}],"version-history":[{"count":8,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/259\/revisions"}],"predecessor-version":[{"id":12975,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/259\/revisions\/12975"}],"wp:attachment":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/media?parent=259"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/categories?post=259"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/tags?post=259"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}