Articles Service
Original
Antigenotoxic and Antimutagenic Effects of Andrographis paniculata , a Traditional Medicinal Herb against Genotoxicity of Cyclophosphamide: An In Vitro Study on Human Peripheral Lymphocytes
1Post-Graduate and Research Department of Biochemistry, Mohamed Sathak College of Arts and Science, University of Madras, Chennai, Tamil Nadu 600119, India 2Department of Zoology, Government Arts College, Periyar University, Salem, Tamil Nadu 636007, India
Prev Nutr Food Sci 2020; 25(3): 246-253
Published September 30, 2020 https://doi.org/10.3746/pnf.2020.25.3.246
Copyright © The Korean Society of Food Science and Nutrition.
Keywords
PDF
Standard view
Export citation
Share Article
Original
Prev Nutr Food Sci 2020; 25(3): 246-253
Published online September 30, 2020 https://doi.org/10.3746/pnf.2020.25.3.246
Copyright © The Korean Society of Food Science and Nutrition.
Antigenotoxic and Antimutagenic Effects of Andrographis paniculata , a Traditional Medicinal Herb against Genotoxicity of Cyclophosphamide: An In Vitro Study on Human Peripheral Lymphocytes
Ayyakkannu Purushothaman1, Parveen Sufiya1, Packirisamy Meenatchi1, Ramalingam Sundaram1, and Nallappan Saravanan2
1Post-Graduate and Research Department of Biochemistry, Mohamed Sathak College of Arts and Science, University of Madras, Chennai, Tamil Nadu 600119, India 2Department of Zoology, Government Arts College, Periyar University, Salem, Tamil Nadu 636007, India
Abstract
Andrographis paniculata (family: Acanthaceae) is a medicinal herb－used in Indian system of medicine (Ayurveda, Siddha, and Unani), traditional and folk systems to treat various illnesses. This study examined the phytochemical constituents of ethanol extract from A. paniculata and its protective effect against genotoxicity caused by cyclophosphamide (CPA). Phytochemical screening and estimation of total phenolic content were analyzed using standard methods. The bioactive components from the ethanol extract of A. paniculata (EAP) were analyzed using gas chromatography-mass spectrometry. To investigate the protective effect of EAP against CPA-induced genotoxicity, human peripheral lymphocyte cultures were used. To test the antigenotoxic and antimutagenic effects of EAP, lymphocytes were treated with different concentrations of extract (50∼250 mg/mL) alone and co-treated along with CPA+EAP for 48 h. The cells were analyzed for structural chromosomal aberrations (CAs) and sister chromatid exchanges (SCEs) in control, CPA treated, and CPA+ EAP co-treated lymphocytes. Results of the study revealed that the lymphocyte cultures which had 48 h continuous exposure to EAP (50∼250 mg/mL) did not show any significant changes in CAs and SCE frequencies. These results substantiate the antimutagenic nature of the extract. Furthermore, the lymphocytes co-treated with CPA along with extract showed a significant reduction in CAs (reduced from 26.50±2.50% to 11.00±1.00%) and SCEs (reduced from 9.92±0.63 per cell to 4.56±0.18 per cell). These results suggest that A. paniculata is protective against CPA induced genotoxicity and put forward its possible use as a supplement with chemotherapeutic drugs.
Keywords: Andrographis paniculata, chromosomal aberrations, cyclophosphamide, genotoxicity, sister chromatid exchange
Fig 1.
Download
Photographs showing
Fig 2.
Download
A representative gas chromatography-mass spectrometry chromatogram of ethanol extract from
-
Table 1 . The phytochemical components in ethanol extract of
Andrographis paniculata based on preliminary phytochemical screening testsNo. Phytochemical constituent Test Inference 1 Alkaloids Wagner’s test ++ Mayer’s test 2 Carbohydrates Molisch’s test + Fehling’s test 3 Flavonoids Shinoda test ++ 4 Glycosides Borntrager’s test ++ Legal’s test Keller-Killiani test 5 Saponins Froth test + 6 Steroids Salkowski reaction +++ 7 Tannins and phenolic compounds Ferric chloride test ++ Lead acetate test Nitric acid test 8 Terpenoids Libermann-Burchard’s test + +, mildly present; ++, highly present; +++, more highly present.
-
Table 2 . Extract yield and total phenolics content for
Andrographis panicuata Sample Extract yield (%) Total phenolics (mg GAE/g) Extract of A. paniculata 15.24±1.76 35.66±3.42 Each value is expressed as mean±SD from minimum of three independent experiments.
GAE, gallic acid equivalent.
-
Table 3 . Major phytochemical components identified by gas chromatography mass spectrometry (GC-MS) analysis from ethanol extract of
Andrographis paniculata Retention time Peak area (%) Name of the compound Library ID 16.448 3.89 Bicyclo[3.1.1]heptanes 2,6,6-trimethyl-(1α,2β,5α) 17013 006876 13 760 Bicyclo[3.1.1]heptane, 2,6,6-trimethyl 16968 000473 55 255 Bicyclo[3.1.1]heptane, 2,6,6-trimethyl-, [1R-(1α,2β,5α)] 17016 004795 86 250 17.320 1.45 Hexadecanoic acid, methyl ester 119400 39 98 Hexadecanoic acid, methyl ester 000112 39 98 Hexadecanoic acid, methyl ester 119406 39 97 000112 119408 000112 17.683 19.79 n -Hexadecanoic acid107549 10 399 n -Hexadecanoic acid000057 10 398 tert -Decanoic acid107548 63 893 000057 84453 000544 19.135 7.45 Phytol 141393 86 799 Phytol 000150 86 790 Phytol 141393 86 774 000150 141395 000150 19.353 15.06 Oleic acid 129338 000112 80 199 9-Octadecenoic acid (E) 129353 000112 79 899 6-Octadecenoic acid 129340 66 898 1000336 19.556 4.04 Octadecanoic acid 131262 000057 11 498 Octadecanoic acid 131261 000057 11 496 Octadecanoic acid 131258 000057 11 496 23.201 13.91 2-Amino-4-morpholino-6-phenylcarbamoyl-1,3,5-triazine 144180 007537 63 553 Ethanone, 1-(4-hydroxy-3,5-dimethoxy phenyl) 57688 002478 38 845 Ethanone, 1-(4-hydroxy-3,5-dimethoxy phenyl) 57687 002478 38 843 24.015 2.36 Anthranilic acid, N-methyl-, butyl ester 66844 015236 34 743 1,2-Benzisothiazol-3-amine tert -butyldimethylsilyl113699 57 238 Hexahydropyridine, 1-methyl-4-(4,5) 1000332 47 138 66899 094427 26.208 4.73 1,2-Benzithiazol-3-amine tert -butyldimethylsilyl113699 57 250 2-(Acetoxymethyl)-3-(methoxycarbonyl) biphenylene 1000332 70 946 5-Methyl-2-phenylindolozine 129190 093103 99 743 67006 036944 28.212 8.18 Stigmasterol 216703 000083 48 756 tert -Butyl(5-isopropyl-2-methylphenoxy) dimethylsilane114128 02 338 Cyclohexane carboxamide, N-furfuryl 1000367 32 838 66823 006341 28.793 28.61 γ-Sitosterol 217434 000083 47 690 β-Sitosterol 217432 000083 46 556 Stigmast-7-en-3-ol, (3β,5α,24S) 217448 018525 35 443
-
Table 4 . The structural chromosomal aberrations (CAs) and percentage of abnormal cells in human lymphocytes treated with cyclophosphamide (CPA) alone,
Andrographis paniculata (EAP) alone and CPA+EAP after 48 h treatmentTreatment CA Structural CA/cell % of abnormal cells Chromatid breakage Chromosome breakage Normal control 11 3 0.07 6.50±0.50 CPA (100 μg/mL) 38 16 0.27** 26.50±2.50** EAP 50 mg/mL 9 4 0.07 6.50±1.50 EAP 100 mg/mL 10 2 0.06 5.50±0.50 EAP 150 mg/mL 12 5 0.08 8.00±1.00 EAP 200 mg/mL 12 6 0.09 8.50±1.50 EAP 250 mg/mL 14 5 0.10 9.00±1.50 CPA+EAP 50 mg/mL 33 12 0.23*# 21.50±1.00*# CPA+EAP 100 mg/mL 29 12 0.21*# 20.50±1.50*# CPA+EAP 150 mg/mL 24 9 0.17**## 15.50±1.50**## CPA+EAP 200 mg/mL 20 9 0.15**## 13.50±1.50**## CPA+EAP 250 mg/mL 17 7 0.12**## 11.00±1.00**## Significantly different from normal control at *
P <0.05 and **P <0.01, and positive control (CPA) at #P <0.05 and ##P <0.01.CAs were evaluated in 200 well spread metaphases per concentration. A total 100 well spread metaphases per donor, no. of donors=2.
-
Table 5 . The SCEs per chromosome and sister chromatid exchanges (SCEs) per cell in human lymphocytes treated with cyclophosphamide (CPA) alone, ethanol extract of
Andrographis paniculata (EAP) alone and CPA+EAP after 48 h treatmentTreatment No. of metaphases scored Total no. of SCEs scored Mean SCEs / chromosome SCEs/cell (mean±SE) Normal control 50 127 0.0552 2.54±0.12 CPA (100 μg/mL) 50 496 0.2156** 9.92±0.63** EAP 50 mg/mL 50 120 0.0521 2.40±0.08 EAP 100 mg/mL 50 124 0.0539 2.48±0.06 EAP 150 mg/mL 50 119 0.0508 2.34±0.06 EAP 200 mg/mL 50 126 0.0547 2.52±0.11 EAP 250 mg/mL 50 131 0.0569 2.62±0.05 CPA+EAP 50 mg/mL 50 419 0.1821*# 8.38±0.14*# CPA+EAP 100 mg/mL 50 376 0.1634*## 7.52±0.11*## CPA+EAP 150 mg/mL 50 310 0.1347**## 6.20±0.13**## CPA+EAP 200 mg/mL 50 274 0.1191**## 5.48±0.20**## CPA+EAP 250 mg/mL 50 228 0.0991**## 4.56±0.18**## Significantly different from normal control at *
P <0.05 and **P <0.01, and positive control (CPA) at #P <0.05 and ##P <0.01.A total 50 well spread metaphases/cultures were scored for the occurrence of SCE, no. of donors=2.
