Abstract Title:

Protective effects of lupeol against mancozeb-induced genotoxicity in cultured human lymphocytes.

Abstract Source:

Phytomedicine. 2016 Jun 15 ;23(7):714-24. Epub 2016 Apr 11. PMID: 27235710

Abstract Author(s):

Amit Kumar Srivastava, Sanjay Mishra, Wahid Ali, Yogeshwer Shukla

Article Affiliation:

Amit Kumar Srivastava


BACKGROUND: Lup-20(29)-en-3H-ol (Lupeol), a dietary pentacyclic triterpenoid has been shown to possess multiple medicinal activities including anti-inflammatory, anti-oxidant and anti-carcinogenic effects. Mancozeb is a widely used broad-spectrum fungicide with well-known carcinogenic hazards in rodents.

PURPOSE: The present study has been designed to investigate the protective effects of lupeol against mancozeb-induced genotoxicity and apoptosis in cultured human lymphocytes (CHLs).

METHODS: The genotoxic effect of mancozeb was evaluated by chromosomal aberration and micronucleus assays. The cell cycle kinetics and intracellular reactive oxygen species (ROS) generation was measured by flow cytometry. The levels of anti-oxidant enzymes and lipid peroxidation (LPO) were estimated by enzymatic assays. The localization of p65NF-κB was measured by immunocytochemical analysis. The differential expression of genes associated with genotoxicity was measured by qRT-PCR.

RESULTS: Mancozeb exposure (5µg/ml) for 24h caused significant induction of chromosomal aberrations (CAs) and micronuclei (MN) formation in CHLs. Pre-and post-treatment (25 and 50µg/ml) of lupeol for 24h significantly (p<0.05) reduced the frequency of CAs and MN induction, in a dose-dependent manner in mancozeb treated CHLs. Concomitantly, lupeol pre-treatment for 24h significantly increased the levels of anti-oxidant enzymes, superoxide dismutase (SOD) and catalase and decreased ROS generation and LPO. Additionally, lupeol pre-treatment significantly reduced mancozeb-induced apoptosis as shown by Sub-G1 peak analysis and annexin V-PI assay, in a dose dependent manner. Moreover, pre-treatment with lupeol attenuated mancozeb-induced NF-κB activation in CHLs. Furthermore, the results of qRT-PCR showed that lupeol pre-treatment significantly (p<0.05) decreased mancozeb-induced expression of DNA damage (p53, MDM2, COX-2, GADD45α and p21) and increased expression of DNA repair responsive genes (hOGG1 and XRCC1) in CHLs.

CONCLUSION: Taken together, our findings suggest that lupeol could attenuate mancozeb-induced oxidative stress, which in turn could inhibit NF-κB activation and thus provide protection against mancozeb-induced genotoxicity and apoptosis. So, lupeol could be used as a potent anti-oxidant regimen against pesticide induced genotoxicity in agricultural farm workers.

Study Type : In Vitro Study

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