Annu Arora

Antimutagenic potential of curcumin on chromosomal aberrations in Wistar rats.

Curcumin, a yellow pigment commonly used as a spice and food coloring agent is obtained from rhizomes of Curcuma longa and is a major chemopreventive component of turmeric. In the present set of investigations the antimutagenic potential of curcumin has been evaluated using in vivo chromosomal aberration assay in Wistar rats. Cyclophosphamide (CP), a well-known mutagen was given by intraperitoneal (i.p.) injection at the dose of 40 mg/kg body weight (b.w.). Curcumin was given at the dose of 100 and 200 mg/kg b.w. through gastric intubation for seven consecutive days prior to CP treatment. The animals were sacrificed at the sampling time of 24 h after treatment and their bone marrow tissue was analyzed for chromosomal damage and mitotic index. In CP treated animals a significant induction of chromosomal aberration was recorded with decrease in mitotic index. However, in curcumin-supplemented animals, no significant induction in chromosomal damage or change in mitotic index was recorded. In different curcumin-supplemented groups, a dose dependent significant decrease in CP induced clastogenicity was recorded. The incidence of aberrant cells was found to be reduced by both the doses of curcumin when compared to CP treated group. The anticytotoxic potential of curcumin towards CP was also evident as the status of mitotic index was found to show increment. The study revealed the antigenotoxic potential of curcumin against CP induced chromosomal mutations.

Carcinogenic and cocarcinogenic potential of cypermethrin on mouse skin

Cypermethrin (CYM), a synthetic pyrethroid insecticide, is used widely because of its high bio-efficacy and low mammalian toxicity. In the present set of investigations, CYM has been evaluated for its carcinogenic and co-carcinogenic (tumour initiating and tumour promoting) potential in mouse skin model of carcinogenesis. The results revealed that CYM possess complete carcinogenic as well as tumour initiating and promoting potential in both the sexes of Swiss albino mice. At the end point, i.e. 32 weeks in a single dose (10 mg/kg body weight (wt.), once only), initiated mice, 9 out of 12 surviving males, and 10 out of 14 surviving females developed benign tumours, while a higher incidence of tumourigenesis was recorded in multiple dose-initiated (10 mg/kg body wt., total nine applications) group, where 7 out of 9 surviving male and 10 out of 13 surviving female mice developed tumours at the site of topical exposure. The application of CYM as a tumour promoter on 7,12-dimethylbenz(a)anthracene initiated animals induced tumour incidence in about 4 out of surviving 10 male and 5 out of surviving 13 female Swiss albino mice. CYM when tested for complete carcinogenic activity induced tumour formation in both male and female animals at all the three tested dose levels.

Induction of apoptosis by diallyl sulfide in DMBA-induced mouse skin tumors.

Diallyl sulfide (DAS), an organosulfur compound in garlic, has received increasing attention as a potential cancer-chemopreventive agent. DAS has been shown to possess antitumorigenic potential in various rodent tumor models. The present study demonstrates induction of apoptosis as the possible mechanism of the antiproliferative effect of DAS in solid tumors, as evident by the appearance of a sub-G1 fraction in flow cytometry. Biochemical analysis of skin tumors revealed other characteristic features of apoptosis, including formation of DNA ladders on agarose gel, compaction of nuclear DNA, and formation of apoptotic bodies. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay of skin tumorsof DAS-supplemented animals showed a significant increase in the number of apoptotic cells compared with animals exposed to 7,12-dimethylbenz[a]anthracene alone. The increase in apoptotic index in nonmalignant and malignant tumors was 78% and 94%, respectively, in DAS-pretreated animals and 68% and 82%, respectively, in animals given DAS 1 h after carcinogen administration. These observations suggest that induction of apoptosis may be the major contributing factor for antitumorigenic properties of DAS.

Suppression of altered hepatic foci development by curcumin in wistar rats.

Curcumin, a yellow pigment of turmeric (Curcuma longa), is a commonly used spice and a coloring agent in foods, drugs, and cosmetics. Curcumin is known to possess chemopreventive properties in various animal tumor models. In the present study the effect of curcumin on the development of altered hepatic foci (AHF), by using a medium term liver bioassay, has been evaluated. AHF were analyzed by quantitative stereology using the Leica Qwin Image Analysis system from frozen liver sections stained for g-glutamyl transferase, adenosine triphosphatase, glucose-6-phosphatase, alkaline phosphatase, and placental isozyme of glutathione S-transferase. A significant protection on diethylnitrosamine (DEN) initiated and 2-acetylaminofluorene (AAF) promoted AHF by curcumin was observed on these biological markers. The curcumin administration was found to restore the normal levels of the enzymes glutathione S-transferase and g-glutamyl transferase in rat liver following DEN-AAF exposure. Similarly, a significant protection was provided by curcumin in the enzyme-deficient foci for the adenosine triphosphatase-, alkaline phosphatase-, and glucose-6-phosphatase-treated groups in comparison to the DEN-AAF-treated group. These results show that curcumin can effectively suppress the DEN-induced development of AHF in rat liver.

Modulation of vinca-alkaloid induced P-glycoprotein expression by indole-3-carbinol

The over-expression of mdr-1 gene transcript P-glycoprotein (P-gp), responsible for multiple drug resistance, is one of the major obstacles in cancer chemotherapy. In the present study, indole-3-carbinol (I3C), a well-known chemopreventive agent present in cruciferous vegetables, has been evaluated for its potential to modulate the over-expression of P-gp induced by vinblastine or vincristine, which are known inducers of mdr-1 gene. The results revealed that I3C significantly reversed the over-expression of P-gp in vinca-alkaloid induced drug resistance as evident by Western blotting using monoclonal antibody (clone JSB1). Quantization of immunostained tissue sections using image analysis technique revealed that vinblastine/ vincristine induced overexpression of P-gp was effectively reversed by I3C. The present investigation suggests that I3C can significantly inhibit the P-gp over-expression and may have utility as a dietary adjuvant in the treatment of cancer for the reversal of multiple drug resistance.

Tumourigenic studies on deltamethrin in Swiss albino mice.

Deltamethrin, an alpha-cyano type II synthetic pyrethroid insecticide is used to control a wide range of insects on a variety of crops. Deltamethrin is reported to cause many adverse effects on non-target species. Deltamethrin is reported to cause DNA damage and micronuclei induction in human lymphocytes. It is highly toxic for other organisms such as aquatic invertebrates, fish and Daphnia. About the tumorigenic risk (both tumour initiating and promoting) associated with deltamethrin exposure, very few reports are available in literature. In the present set of investigations, deltamethrin has been evaluated for its tumorigenic and co-carcinogenic (tumour initiating and tumour promoting) potential following long term dermal exposure in Swiss albino mice. The results revealed that deltamethrin has only tumour initiating potential in both the sexes of Swiss albino mice, initiated with deltamethrin and promoted by standard tumour promoter, 12-O-tetra decanoyl phorbol-13-acetate (TPA). In the single dose initiated mice (deltamethrin 4 mg/kg body weight, once only), 44% males and 43% females developed benign skin tumours. A much higher incidence of tumorigenesis was recorded in multiple dose initiated animals (deltamethrin 4 mg/kg body weight, three times per week for 3 weeks), where 71% male and 75% female mice developed tumours at the site of application of deltamethrin. Deltamethrin exposure failed to show any tumour promoting and complete tumorigenic potential at all the three tested dose levels.

Chemopreventive effect of indole-3-carbinol on induction of preneoplastic altered hepatic foci.

Abstract: Indole-3-carbinol (I3C) is a cleavage product of glucobrassicanin, a natural compound present in a wide variety of plant food substances including members of the family Cruciferae. I3C is known to possess cancer-chemopreventive potential in various animal models. The present study reveals the protective effect of I3C on the development of diethylnitrosamine (DEN)-initiated and 2-acetylaminofluorene (AAF)-promoted preneoplastic, altered hepatic foci (AHF) in Wistar rats. I3C was given at dose levels of 0.5 and 1 mg/kg body weight for five consecutive days along with DEN and AAF. AHF were scored and analyzed by quantitative stereology using the Image Analysis System from frozen liver sections stained for positive and negative biological markers of AHF, that is, glutathione S-transferase (GST-P), γ-glutamyl transpeptidase (GGT), glucose-6-phosphatase (G6Pase), adenosine triphosphatase (ATPase), and alkaline phosphatase (AlkPase). Results revealed the chemopreventive effect of I3C on the DEN-initiated AHF in Wistar rats. The expression of G6Pase, ATPase, and AlkPase was restored in the I3C-supplemented animal. Similarly the induced expression GST-P and GGT also decreased in the animals with I3C administration. The recovery of altered levels of these biomarkers was of comparatively higher magnitude in the animals given a higher dose of I3C (1 mg/kg body weight) in comparison with the animals given 0.5 mg/kg body weight dose of I3C, although no dose-dependence pattern was recorded in I3C-supplemented groups. These results thus suggest the chemopreventive effect of I3C in rat hepatocarcinogenesis by suppressing DEN- and AAF-induced AHF development.

Enhancing effects of mustard oil on preneoplastic hepatic foci development in Wistar rats

Dietary habits are known to be the major contributory factor in the development of cancer. Mustard oil, which is extensively used in India and elsewhere as a frying and cooking medium, is reported to induce an inflammatory response. The development of altered hepatic foci is an early carcinogenic change in rat liver in diethylnitrosamine (DEN)-induced hepatocarcinogenesis. In the present study, the development of preneoplastic lesions was observed following administration of mustard oil (0.5 mL/day for 8 weeks) in DEN-initiated and partially hepatomized Wistar rats. A significant decrease in the relative and absolute liver weight of mustard oil-exposed rats was recorded. The results revealed a significant increase in the number and area of placental glutathione S-transferase (GST-P) and g-glutamyl transpeptidase (GGT)-positive foci in mustard oil-administered animals. The GST-P and GGT-positive foci were more prominent in the animals given boiled (up to 3008C for 3 hours) mustard oil in comparison to the animals given fresh mustard oil. These results indicate the possible tumourigenic risk associated with mustard oil consumption.

Protective effects of indole-3-carbinol on cyclophosphamide-induced clastogenecity in mouse bone marrow cells

Indole-3-carbinol (I3C) is present in many cruciferous vegetables and is known to possess protective properties against chemically induced toxicity and carcinogenesis. In the present study, the antimutagenic potential of I3C has been evaluated using in vivo chromosomal aberration (CA) assay as a cytogenetic end point. Chromosomal analysis was carried out in mouse bone marrow cells following administration of I3C (5 mg/kg; i.p.) for 5 consecutive days. Cyclophosphamide (CP), a well known mutagen, was given at two dose levels of 25 mg/kg b.wt. and 100 mg/kg b.wt., respectively, 24 hours prior to the last dose of I3C. Two groups of five mice each were also injected with CP (25 or 100 mg/kg b.wt.) alone whereas for the vehicle control a group of mice was injected with normal saline only. The results revealed a significant inhibition in the frequencies of CP-induced CAs and aberrant cells in bone marrow cells of I3C-supplemented Swiss albino mice. The antimutagenic potential of I3C towards CP was also evident as the status of mitotic index (MI) was found to show an increment. This study revealed the antigenotoxic potential of I3C against CP- induced chromosomal mutations.

Induction of preneoplastic altered hepatic foci following dietary sulphur supplementation

Sulphur is an essential micronutrient required by the body in low concentrations, but its high intake can lead to a serious health hazard. Sulphur compounds are reported to induce several toxic responses in animals, but so far no reports are available on the toxic effects of elemental sulphur, following dietary supplementation. The present investigation was carried out with the aim of providing an insight into the role of dietary supplementation of sulphur on the induction of altered hepatic foci (AHF) using medium term liver bioassay in Wistar rats. Induction of AHF are early neoplastic changes in rat liver in diethylnitrosamine (DEN)-initiated and 2-acetylamino fluorene (2-AAF)-promoted hepatocarcinogenesis. The role of sulphur on induction of AHF was evaluated by the development of negative enzymatic foci for alkaline phosphatase (AlkPase), adenosine triphosphatase (ATPase), glucose-6-phosphatase (G-6-Pase) and positive foci for marker enzymes, glutamyl transferase (GGT), placental isozyme of glutathione-S transferase (GST-P). A significant dose-dependent decrease in the relative and absolute liver weight of sulphur-administered rats was recorded. Dietary supplementation of 2% and 4% sulphur significantly induces both negative and positive focal areas in terms of area and counts for AHF. However, 1% sulphur administration failed to induce AHF up to significant levels. The results thus revealed the possible tumorigenic risk associated with the high sulphur-containing diet.