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Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (12): 1433-1442.doi: 10.3736/jcim20121216

• Original Experimental Research • Previous Articles     Next Articles

Dihydroxy-isosteviol methyl ester of Pulsatilla nigricans extract reduces arsenic-induced DNA damage in testis cells of male mice: Its toxicity, drug-DNA interaction and signaling cascades

Samadder Asmita1,Das Jayeeta1,Das Sreemanti1,Das Durba1,De Arnab1,Bhadra Kakali2,Rahman Khuda-Bukhsh Anisur1()   

  1. 1. Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India
    2. Entomology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India
  • Received:2012-05-17 Accepted:2012-06-19 Online:2012-12-20 Published:2018-12-15

Objective: To evaluate the ameliorative efficacy of dihydroxy-isosteviol methyl ester (DIME) of Pulsatilla nigricans extract in arsenic-induced DNA damage in testis cells of mice.
Methods: The mice were treated with sodium arsenite (SA) solution intragastrically at a dose of 20 mg/kg per day and examined at 30, 60, and 90 d after treatment. We divided SA-intoxicated mice into two sub-groups: one fed with DIME at a dose of 35 mg/kg and the other with 85% alcohol. We analyzed the expressions of apoptotic signal proteins like CYP1A1, p53 and caspase 3, ascertained the level of cellular and DNA damage and estimated the level of testicular-toxicity biomarkers. We studied the interaction of DIME with calf thymus DNA as target through circular dichroism spectra and melting temperature profiles.
Results: We observed an elevation in all apoptotic and toxicity biomarkers leading to cellular and DNA damage in the SA-intoxicated mice which showed significant inhibition or reversal on administration of DIME. Results also showed that DIME interacted with DNA, bringing in discernible changes in structure and conformation.
Conclusion: DIME has potentials for therapeutic use in amelioration of arsenic-induced reproductive toxicity.

Key words: Pulsatilla, plant extracts, arsenites, apoptosis, spermatids, mice

Figure 1

Probable chemical structure of dihydroxy-isosteviol methyl ester."

Figure 2

Bar diagrams of activities of ACP and ALP at different time intervals Data are presented as mean±standard error of mean, n=3; *P<0.05, **P<0.01, vs SA plus placebo-treated group. ACP: acid phosphatase; ALP: alkaline phosphatase; SA: sodium arsenite; DIME: dihydroxy-isosteviol methyl ester."

Figure 3

Bar diagrams showing sperm count at different intervals of treatment Data are presented as mean±standard error of mean, n=3; *P<0.05, **P<0.01, vs SA plus placebo-treated group. SA: sodium arsenite; DIME: dihydroxy-isosteviol methyl ester."

Figure 4

Bar diagrams showing arsenic content in testis at different time intervals Data are presented as mean±standard error of mean, n=3; *P<0.05, **P<0.01, vs SA plus placebo-treated group. SA: sodium arsenite; DIME: dihydroxy-isosteviol methyl ester."

Figure 5

Images showing sperm head morphology (Light microscopy, ×200) A: Normal; B-D: SA-intoxicated (30, 60 and 90 d, respectively); E-G: SA plus placebo-treated (30, 60 and 90 d, respectively); H-J: SA plus DIME-treated (30, 60 and 90 d, respectively). SA: sodium arsenite; DIME: dihydroxy-isosteviol methyl ester."

Figure 6

DNA fragmentation assessment Ln1: Normal; Ln2: SA-intoxicated; Ln3: SA plus placebo-treated; Ln4: SA plus DIME-treated. SA: sodium arsenite; DIME: dihydroxy-isosteviol methyl ester."

Figure 7

CD spectra of calf-thymus DNA in the absence and presence of DIME CD: circular dichroism; DIME: dihydroxy-isosteviol methyl ester."

Figure 8

Graphs of DNA thermal denaturation DIME: dihydroxy-isosteviol methyl ester; OD: optical density."

Figure 9

Immunohistochemical staining for p53 in testes (Light microscopy, ×200) A: Normal; B-D: SA-intoxicated (30, 60 and 90 d, respectively); E-G: SA plus placebo-treated (30, 60 and 90 d, respectively); H-J: SA plus DIME-treated (30, 60 and 90 d, respectively). SA: sodium arsenite; DIME: dihydroxy-isosteviol methyl ester."

Figure 10

Photographs of sperm cells stained with DAPI (Fluorescence microscopy, ×200) A: Normal; B-D: SA-intoxicated (30, 60 and 90 d, respectively); E-G: SA plus placebo-treated (30, 60 and 90 d, respectively); H-J: SA plus DIME-treated (30, 60 and 90 d, respectively). DAPI: 4′,6-diamidino-2-phenylindole dihydrochloride; SA: sodium arsenite; DIME: dihydroxy-isosteviol methyl ester."

Figure 11

Bands obtained in Western blot and RT-PCR analyses A: Western blot; B: RT-PCR. G3PDH: glyceraldehyde-3-phosphate dehydrogenase; RT-PCR: reverse transcriptase-polymerase chain reaction."

[1] Martinez VD, Vucic EA, Becker-Santos DD, Gil L, Lam WL . Arsenic exposure and the induction of human cancers.J Toxicol. 2011 ; 2011 : 431287.
doi: 10.1155/2011/431287 pmid: 3235889
[2] Sarkar M, Chaudhuri GR, Chattopadhyay A, Biswas NM . Effect of sodium arsenite on spermatogenesis, plasma gonadotrophins and testosterone in rats. AsianJ Androl. 2003; 5(1):27-31.
[3] Cowperthwaite AC . A text-book of materia medica: characteristic, analytical, and comparative. 4th ed. Chicago: Gross & Delbridge. 1887.
[4] Vaughan JG, Judd PA, Bellamy D. The Oxford book of health foods. New York: Oxford University Press. 2003.
[5] Sudheera AP. Pulsatilla nigricans — A comprehensive study.[ 2012-05-02] . Pulsatilla nigricans — A comprehensive study. [2012-05-02] . .
[6] Morakinyo AO, Adeniyi OS, Arikawe AP . Effects of Zingiber officinale on reproductive functions in the male rat. AfrJ Biomed Res. 2008; 11(3):329-334.
[7] Sanghamitra S, Hazra J, Upadhyay SN, Singh RK, Amal RC . Arsenic induced toxicity on testicular tissue of mice. IndianJ Physiol Pharmacol. 2008; 52(1) : 84-90.
pmid: 18831356
[8] Samadder A, Chakraborty D, De A, Bhattacharyya SS, Bhadra K, Khuda-Bukhsh AR . Possible signaling cascades involved in attenuation of alloxan-induced oxidative stress and hyperglycemia in mice by ethanolic extract of Syzygium jambolanum : drug-DNA interaction with calf thymus DNA as target. EurJ Pharm Sci. 2011 ; 44(3):207-217.
[9] Sinha M, Manna P, Sil PC . Protective effect of arjunolic acid against arsenic-induced oxidative stress in mouse brain.[J] Biochem Mol Toxicol. 2008; 22(1):15-26.
doi: 10.1002/jbt.20209 pmid: 18273903
[10] Pintado B, de la Fuente J, Roldan ER . Permeability of boar and bull spermatozoa to the nucleic acid stains propidium iodide or Hoechst 33258, or to eosin: accuracy in the assessment of cell viability.J Reprod Fertil. 2000 ; 118(1):145-152.
[11] Wyrobek AJ, Watchmaker G, Gordon L. Handbook of mutagenecity testing protocol. In: Kilbey BJ, Legator M,Nichols W, Ramel C. Handbook of mutagenecity test procedures. Amsterdam: Elsevier. 1983: 733-750.
[12] Khuda-Bukhsh AR, Pathak S, Guha B, Karmakar SR, Das JK, Baneijee P, Biswas SJ, Mukheijee P, Bhattachaijee N, Choudhury SC, Banerjee A, Bhadra S, Mallick P, Chakrabarti J, Mandal B . Can homeopathic arsenic remedy combat arsenic poisoning in humans exposed to groundwater arsenic contamination? A preliminary report on first human trial. Evid Based Complement Alternat Med. 2005; 2(4) : 537-548.
doi: 10.1093/ecam/neh124
[13] Walter K, Schutt C. Acid and alkaline phosphatase in serum (Two point method). In: Bergmeyer HU. Methods in enzymatic analysis. New York: Academic Press. 1974 : 856-860.
[14] Ramakrishnan G, Raghavendran HR, Vinodhkumar R, Devaki T . Suppression of N-nitrosodiethylamine induced hepatocarcinogenesis by silymarin in rats. Chem Biol Interact. 2006; 161(2):104-114.
doi: 10.1016/j.cbi.2006.03.007 pmid: 16643877
[15] Aman RP . A critical review of methods for evaluation of spermatogenesis from seminal characteristics.J Androl. 1981; 2:37-38.
doi: 10.1002/j.1939-4640.1981.tb00595.x
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