Radiomimetic drug - ‘Bleomycin' induced DNA damage repair by Alstonia scholaris bark extracts – A G2 assay based evidence

Authors

  • Sumitra Chakraborty Dept of Zoology, Sheth M. N. Science College, Patan, Gujarat, India
  • Meonis Pithawala C G Bhakta Institute of Biotechnology, UKA TARSADIA UNIVERSITY, Gopal Vidyanagar, Bardoli Mahua Road, Tarsadi-394350, Gujarat, India

Abstract

Objective: The present study initiated to know the probable DNA damage repair by A. scholaris extracts using Bleomycin-induced chemosensitive G2 assay in in-vitro cultured human blood lymphocytes.

Methods: The plant extracts were from bark, stem and leaves and extraction solvents used were double distilled water and methanol. The experiments were carried out with blood samples collected from 12 healthy volunteers. A total of 28 culture vials from each individual were set up for lymphocyte cultures. These culture tubes (vials) were divided into two groups of fourteen each. The first group was labeled as ‘G0' while the second as ‘G2'. In both groups, initial vials (‘A') were kept untreated so as to act as a control, the second vial labeled (‘B') received treatment of an alone dose of BLM (15μg/ml), while vials labeled ‘C' to ‘N' were treated with varied combinations. At the 71st hour, the cells were treated with 100 μl of 0.001% colchicine. The cultures were terminated at the 72nd hour of incubation. Routine air-dried preparations were made. Each slide was blind coded, conveniently stained in 2% Giemsa and scored under 100X oil immersion lens.

Results: The results indicated that aqueous as well as methanolic bark extracts had enough potential to repair DNA damage caused by Bleomycin. The extracts (both aqueous as well as methanolic) from stem or leaves were not that promising.   

Author Biographies

Sumitra Chakraborty, Dept of Zoology, Sheth M. N. Science College, Patan, Gujarat, India

Assistant Professor, Dept of Zoology

Meonis Pithawala, C G Bhakta Institute of Biotechnology, UKA TARSADIA UNIVERSITY, Gopal Vidyanagar, Bardoli Mahua Road, Tarsadi-394350, Gujarat, India

Associate Professor,

References

Nadkarni A. Indian Materia Medica. 3rd ed. Popular Book Depot: Mumbai, India, 2014.p.243

Joshi SG. Cesalpinaceae. In: Medicinal plants, Oxford and IBH publishing Co. Pvt. Ltd, New Delhi, India, 2000.p.119.

Dutt AT. Total alkaloids in Alstonia scholaris. Sci Cult. 1944; 9: 555-556.

Boonchuay W, Court WE. Alkaloids of Alstonia scholaris from Thailand. Planta Medica. 1976; 29(4): 380-390.

Rahman AU, Alvi KA. Indole alkaloids from Alstonia scholaris. Phytochemistry. 1987; 26(7): 2139-2142.

Kam TS, Nyeoh KT, Sim KM, Yoganathyan K. Alkaloids from Alstonia scholaris. Phytochemistry. 1995; 45(6): 1303-1305.

Hadi S, Bremner JB. Initial studies on alkaloids from Lombok medicinal plants. Molecules. 2001; 6(2):117-129.

Mahidol C, Prawat H, Prachyawarakorn V, Ruchirawat S. Investigation of some bioactive Thai medicinal plants. Phytochemistry Reviews. 2002; 1(3): 287-297.

Dai YY, Yang W, He R, Zhou FL, Mao YL. Resource status, seed germination characteristics and alkaloids contents of Alstonia scholaris in Yunnan. Journal of West China Forestry Science. 2008; 1(1): 73-76.

Cai XH, Shang JH, Feng T, Luoa XD. Novel alkaloids from Alstonia scholaris. Zeitschrift für Natur for schung. 2010; 65b: 1164-1168.

Jain L, Pandey MB, Singh S, Singh AK, Pandey VB. A new indole alkaloid from Alstonia scholaris. Natural Product Research. 2009a; 23(17):1599-1602.

Jain L, Singh S, Pandey MB, Pandey VB. Alkaloids of Alstonia scholaris. Journal of Indian Chemical Society. 2009b; 86: 758-760.

Khyade MS, Kasote DM, Vaikos NP. Alstonia scholaris (L.) R. Br. and Alstonia macrophylla Wall. ex G. Don: A comparative review on traditional uses, phytochemistry and pharmacology. Journal of Ethnopharmacology. 2014; 153(1): 1–18.

Chauhan JS, Chaturvedi R, Kumar S. Isookanin-7-O-Alpha-LRhamnopyranoside: a new flavanone glycoside from the Roots of Alstonia scholaris. Indian Journal of Chemistry – Section B Organic and Medicinal Chemistry. 1985; 24(2): 219.

Thomas PS, Kanaujia A, Ghosh D, Duggar R, Katiyar CK. Alstonoside, a secoiridoid glucoside from Alstonia scholaris. Indian Journal of Chemistry. 2008; 47B: 1298-1302.

Khyade MS, Vaikos NP. Pharmacognostical studies on the leaves of Alstonia scholaris R. Br. Journal of Pharmacy Research. 2009; 2(5): 858–861

Gupta RS, Sharma R, Sharma A, Bhatnager AK, Dobhal MP, Joshi YC, et al. Effect of Alstonia scholaris bark extract on testicular function of Wistar rats. Asian Journal of Andrology. 2002; 4(3): 175-178.

Dung NX, Ngoc RH, Rang DD, Nhan NT, Klinkby N, Leclercq PA. Chemical composition of the volatile concentrate from the flowers of Vietnamese Alstonia scholaris (L.) R. Br., Apocynaceae. Journal of Essential Oil Research. 2001; 13(6): 424-426.

Singh SK, Singh A. Toxic effect of Alstonia scholaris plant to fingerlings of Labeo rohita (Hamilton) in different conditions. World Journal of Zoology. 2010; 5(1): 41-46.

Kirtikar KR, Basu BD. Indian Medicinal Plants vol. II. Indian Press, Allahabad, India: Sudhindra Nath Basu M.B.: 1918. pp. 786–790.

Perry LM, Metzger J. Medicinal Plants of East and Southeast Asia. Cambridge, Mass: MIT Press: 1980.

Burkill IM. A Dictionary of Economic Products of the Malay Peninsula. 2nd ed. Ministry of Agriculture and Co-operatives, Kuala Lumpur, Malaysia: 1966.

Compiling Group of Yunnan Traditional Chinese Medicine. Yunnan Traditional Chinese Medicinal Plant. Yunnan People's Press: Kunming: 1977.

Changwichit K, Khorana N, Suwanborirux K, Waranuch N, Limpeanchob N, Wisuitiprot W, et al. Bisindole alkaloids and secoiridoids from Alstonia macrophylla Wall. ex G. Don. Fitoterapia. 2011; 82(6): 798–804.

Gandhi M, Vinayak VK. Preliminary evaluation of extracts of Alstonia scholaris bark for in vivo antimalarial activity in mice. Journal of Ethnopharmacology. 1990; 29(1): 51-57.

Keawpradub N, KirbyKirby GC, Steele JCP, Houghto PJ. Antiplasmodial activity of extracts and alkaloids of three Alstonia species from Thailand. Planta Medica. 1999; 65 (8): 690-694.

Misra CS, Pratyush K, Lipin Dev MS, James J, Veettil AT, Thankamani V. A comparative study on phytochemical screening and antibacterial activity of roots of Alstonia scholaris with the root, leaves and stem barks. International Journal of Research in Phytochemistry and Pharmacology. 2011; 1(2): 77–82.

Singh B, Sangwan P. Taxonomy, ethnobotany and antimicrobial activity of Alstonia scholaris (L.) R. Br., Carissa carandas L. and Catharanthus roseus (L.) G. Don. International Journal of Biotechnology and Bioscience. 2011; 1(1): 102–112.

Khan MR, Omoloso AD, Kihara M. Antibacterial activity of Alstonia scholaris and Leeatetramera. Fitoterapia. 2003; 74(7-8): 736–740.

Hussain A, Zaman MKA, Ramteke M. Antibacterial activity of trunk bark of Alstonia scholaris. Asian Journal of Pharmaceutical and Clinical Research. 2010; 3(4): 46–47.

Shah AJ, Gowani SA, Zuberi AJ, Ghayur MN, Gilani AH. Antidiarrheal and spasmolytic activities of the methanolic crude extract of Alstonia scholaris L. are mediated through calcium channel blockade. Phytotherapy Research. 2010; 24(1): 28–32.

Arulmozhi S, Mazumder PM, Ashok P, Narayanan LS. In Vitro Antioxidant and free radical scavenging activity of Alstonia scholaris Linn. R. Br. Iranian Journal of Pharmacology and Therapeutics. 2007a; 6(2): 191–196.

Ravi Shankar K, Ramesh KVRNS, Naveena P. Free radical scavenging activity of the flower and fruit extracts of Alstonia scholaris. Biosciences Biotechnology Research Asia. 2008; 5(1): 493–494.

Kumar A, Kaur R, Arora S. Free radical scavenging potential of some Indian medicinal plants. Journal of Medicinal Plants Research. 2010; 4(19): 2034-2042.

Arulmozhi S, Mazumder PM, Narayanan LS, Thakurdesai PA. In vitro antioxidant and free radical scavenging activity of fractions from Alstonia scholaris Linn. R. Br. International Journal of Pharm Tech Research. 2010; 2(1): 18–25.

Anurakkun NJ, Bhandari MR, Kawabata J. α-Glucosidase inhibitors from Devil tree (Alstonia scholaris). Food Chemistry. 2007; 103(4): 1319-1323.

Jagetia GC, Baliga MS. Evaluation of anticancer activity of the alkaloid fraction of Alstonia scholaris (Sapthaparna) in vitro and in vivo. Phytotherapy Research. 2006; 20(2): 103–109.

Sharma V, Mallick SA, Tiku AK. Anticancer activity of Devil tree (Alstonia scholarisLinn.) leaves on human cancer cell lines. Indian Journal of Agricultural Biochemistry. 2010; 23(1): 63–65.

Kamarajan P, Sekar N, Mathuram V, Govindasamy S. Antitumor effect of echitamine chloride on methyl cholonthrene induced fibrosarcoma in rats. Biochemistry International. 1991; 25(3): 491-498.

Arulmozhi S, Mazumder PM, Sathiyanarayanan L, Thakurdesai PA. Analgesic, anti-inflammatory and anti-ulcerogenic activities of fractions from Alstonia scholaris. Pharmacologia. 2012; 3(5): 132-137.

Karawya MS, Ammar NM, Hifnawy MS, Al-okbi SY, Doha AM, El-Anssary AA. Phytochemical study and evaluation of the anti-inflammatory activity of some medicinal plants growing in Egypt. Medical Journal of Islamic World Academy of Sciences. 2010; 18(4): 139-150.

Kulkarni MP, Juvekar AR. Effect of Alstonia scholaris (Linn.) R. Br. on stress and cognition in mice. Indian Journal of Experimental Biology. 2008; 47(1): 47-52.

Bhogayata K, Sharma PP, Patel BR. A clinical evaluation of Saptaparna (Alstonia scholaris L.R. Br.) on essential hypertension. AYU (An International Quarterly Journal of Research in Ayurveda). 2009; 30(3): 318-322.

Arulmozhi S, Rasal VP, Narayanan LS, Ashok P. Screening of Alstonia scholaris Linn. R. Br. for wound healing activity. Oriental Pharmacy and Experimental Medicine. 2007b; 7(3): 254-260.

Lin SC, Lin CC, Lin YH, Supriyatna S, Pan SL. The protective effect of Alstonia scholaris R. Br. on hepatotoxin-induced acute liver damage. American Journal of Chinese Medicine. 1996; 24(2): 153-164.

Iwo MI, Soemardji AA, Retnoningrum DS, Sukrasno UM. Immunostimulating effect of pule (Alstonia scholaris L.R. Br., Apocynaceae) bark extracts. Clinical Hemorheology and Microcirculation. 2000; 23(2-4): 177-183.

Shang JH, Cai XH, Zhao YL, Feng T, Luo XD. Pharmacological evaluation of Alstonia scholaris: anti-tussive, anti-asthmatic and expectorant activities. Journal of Ethnopharmacology. 2010; 129(3): 293-298.

Gupta U, Jahan S, Chaudhary R, Goyal PK. Amelioration of radiation-induced haematological and biochemical alterations by Alstonia scholaris (a medicinal plant) extract. Integrative Cancer Therapies. 2008; 7(3):155-161.

Gupta U, Chaudhary R, Goyal PK. Post-treatment effects of Alstonia scholaris extract against radiation induced biochemical alterations in swiss albino mice. Iranian Journal of Radiation Research. 2010; 8(3): 169-177.

Jahan S, Goyal PK. Protective effect of Alstonia scholaris against radiation-induced clastogenic and biochemical alterations in mice. Journal of Environmental Pathology, Toxicology, and Oncology.2010; 29(2): 101-111.

Handa SS, Khanuja SPS, Longo G, Dev-Dutt R. Extraction Technologies for Medicinal and Aromatic Plants. 1st ed. Italy: United Nations Industrial Development Organization and the International Centre for Science and High Technology: 2008.

Hungerford DA. Leukocytes cultured from small inocula of whole blood and preparations of metaphase chromosomes by treatment with hypotonic KCl. Stain Technology. 1965; 40(6): 333-338.

Painuli S, Kumar N. Prospects in the development of natural radioprotective therapeutics with anti-cancer properties from the plants of Uttarakhand region of India. Journal of Ayurveda and Integrative Medicine. 2016; 7(1): 62–68.

Kopjar N, Mioèiæ S, Ramiæ S, Miliæ M, Viculin T. Assessment of the radioprotective effects of amifostine and melatonin on human lymphocytes irradiated with gamma rays in vitro. Archives of Industrial Hygiene and Toxicology. 2006; 57(2): 155-163.

Copeland ES. Mechanisms of radioprotection-a review. Photochemistry and Photobiology. 1978; 28(4-5): 839-844.

Uma Devi P. Normal tissue protection in cancer therapy: progress and prospects. Acta Oncologica.1998; 37(3): 247–252.

Maurya DK, Devasagayam PA, Nair CKK. Some novel approaches for radioprotection and the beneficial effect of natural products. Indian Journal of Experimental Biology. 2006; 44 (2): 93-114.

Maurya DK, Balakrishnan S, Salvi VP, Nair CKK. Protection of cellular DNA from γ-radiation-induced damages and enhancement in DNA repair by troxerutin. Molecular and Cellular Biochemistry. 2005a; 280(1-2): 57-68.

Maurya DK, Salvi VP, Nair CKK. Radiation protection of DNA by ferulic acid under in vitro and in vivo conditions. Molecular and Cellular Biochemistry. 2005b; 280 (1-2): 209-217.

Vasin MV. Comments on the mechanisms of action of radiation protective agents: basis components and their polyvalence. Springer Plus. 2014; 3(1): 414.

Jahan S, Goyal PK. Protective effect of Alstonia scholaris against radiation-induced clastogenic and biochemical alterations in mice. Journal of Environmental Pathology, Toxicology, and Oncology.2010; 29(2): 101-111.

Gupta U, Jahan S, Chaudhary R, Goyal PK. Amelioration of radiation-induced haematological and biochemical alterations by Alstonia scholaris (a medicinal plant) extract. Integrative Cancer Therapies. 2008; 7(3):155-161.

Gupta U, Chaudhary R, Goyal PK. Radio-response to leucocytes in peripheral blood of mice against gamma irradiation and their protection by Alstonia scholaris extract. Nuclear Technology & Radiation Protection. 2011; 26(2): 126-133.

Gupta U, Agrawal NK, Chaudhary R, Goyal PK. Radioprotective role of Alstonia Scholaris extract against hematological dysfunctions in mice. Journal of Radioprotection Research. 2013; 1(1): 1-9.

Hussain A, Gogoi B, Ramteke A. Protective effects of Alstonia scholaris (L.) R. Br. bark extract against oxidative stress induced by Hydrogen Peroxide. Pharmacognosy Communications. 2013; 3(1): 16-20.

Jagetia GC, Baliga MS. Treatment with Alstonia scholaris enhances radiosensitivit in vitro and in vivo. Cancer Biotherapy and Radiopharmaceuticals. 2003;18(6): 917-929.

Jahan S, Goyal P, editors. Prevention of radiation-induced chromosomal aberrations in bone marrow of mice by Indian medicinal plant, Alstonia scholaris. DAE-BRNS life sciences symposium 2007 on DNA damage, repair and their implications: proceedings: 2007.

Hutchinson F. Sulfhydryl groups and the oxygen effect on irradiated dilute solutions of enzymes and nucleic acids. Radiation Research. 1961; 14(6): 721-731.

El-Zein RA, Monroy CM, Cortes A, Spitz MR, Greisinger A, Etzel CJ. Rapid method for determination of DNA repair capacity in human peripheral blood lymphocytes amongst smokers. BMC Cancer. 2010; 10(1): 439.

Samarth RM. Protection against radiation induced hematopoietic damage in bone marrow of Swiss albino mice by Mentha piperita (Linn). Journal of Radiation Research. 2007; 48(6): 523-528.

Silva JP, Gomes AC, Coutinho OP. Oxidative DNA damage protection and repair by polyphenolic compounds in PC12 cells. European Journal of Pharmacology. 2008; 601(1): 50-60.

Sisodia R, Singh S, Sharma K, Ahaskar M. Post treatment effect of Grewia asiatica against radiation-induced biochemical alterations in Swiss albino mice. Journal of Environmental Pathology, Toxicology and Oncology. 2008; 27(2): 113-121.

Sharma KV, Sisodia R. Evaluation of the free radical scavenging activity and radioprotective efficacy of Grewia asiatica fruit. Journal of Radiological Protection. 2009; 29(3): 429-443.

Li A, Schuermann D, Gallego F, Kovalchuk I, Tinland B. Repair of damaged DNA by Arabidopsis cell extract. The Plant Cell. 2002; 14(1): 263-273.

Published

01-03-2019

How to Cite

Chakraborty, S., & Pithawala, M. (2019). Radiomimetic drug - ‘Bleomycin’ induced DNA damage repair by Alstonia scholaris bark extracts – A G2 assay based evidence. Innovare Journal of Life Sciences, 7(2). Retrieved from https://journals.innovareacademics.in/index.php/ijls/article/view/29462

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