Azoxymethane Group Research Articles

Morphometric investigation of the effects of Azoxymethane, Diallyl disulfide and corn oil use on humerus and femur development in rats

The study aimed to examine the effects of Azoxymethane (AOM), Diallyl Disulfide (DADS), and corn oil on the humerus and femur in rats. In the study, 40 male Wistar Albino rats, 12 weeks old, were used. The animals were divided into four different groups (Control, AOM, DADS and Corn oil). After the experimental period, all animals were anesthetized and sacrificed by cervical dislocation. Humerus and femur from long bones resected from all animal groups. The maximum length, proximal width, distal width, diaphysis diameter, cortex thickness and cavum medullare diameter of the bones were measured. After the morphometric measurements index 1, index 2, robusticity index, and bone weight/bone length index were calculated for the bones. No difference was observed in the morphometric measurements and indices performed on the humerus. A statistical difference in proximal width and robusticity index was detected between the groups in the femur. A difference was observed between the AOM group and the Control/corn oil groups in proximal width. A difference was detected between the AOM group and the Control group in the robustness index, and it was determined that AOM caused a decrease in density and strength in bones. It is thought that the findings obtained in this study will contribute to the evaluation of the effects of cancer research using AOM on bones in rats.

Investigation into the Potential Role of Propionibacterium freudenreichii in Prevention of Colorectal Cancer and Its Effects on the Diversity of Gut Microbiota in Rats.

Colorectal cancer (CRC) accounts for 10% of all cancer diagnoses and cancer-related deaths worldwide. Over the past two decades, several studies have demonstrated the clinical benefits of probiotic supplementation and some studies have shown that certain probiotics can modulate immunity and strengthen gut microbiota diversity. This study aims to assess the impact of the Propionibacterium freudenreichii (PF) probiotic against CRC induced by azoxymethane (AOM), and to investigate its effects on gut microbiota diversity in rats, as well as to evaluate the anti-proliferative activities of PF in HCT116 CRC cells. This experiment was performed using four groups of SD rats: normal control, AOM group, PF group (1 × 109 CFU/mL), and standard drug control (5-fluorouracil, 35 mg/kg). Methylene blue staining of colon tissues showed that the administration of PF significantly reduced the formation of colonic aberrant crypt foci (ACF) compared to the AOM control group. In addition, treated rats had lower levels of malondialdehyde in their colon tissue homogenates, indicating that lipid peroxidation was suppressed by PF supplementation. Furthermore, 16S rRNA gene analysis revealed that probiotic treatment enhanced the diversity of gut microbiota in rats. In vitro study showed that the viability of HCT116 cells was inhibited by the probiotic cell-free supernatant with an IC50 value of 13.3 ± 0.133. In conclusion, these results reveal that consuming PF as probiotic supplements modulates gut microbiota, inhibits the carcinogenic effects of AOM, and exerts anti-proliferative activity against CRC cells. Further studies are required to elucidate the role of PF on the immune response during the development and growth of CRC.

Palm Oils Consumption Modulates Serum SGPT and SGOT in Rats Fed a High-Fat Diet

In this study we investigated the effect of three kinds of palm oil product (refined bleached deodorized palm oil (RBDPO), red palm oil (RPO) and palm kernel oil (PKO)) on the serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), and liver color of rats in fed high-fat diet, and compare it with beef tallow and corn oil. Growing male SD rats were fed a high-fat diet (20% fat) for 13 weeks, divided into 2 groups (with and without azoxymethane (AOM)). Dietary corn oil showed the lowest serum SGOT and SGPT (p < 0.05) both in the AOM and Non-AOM groups. Beef tallow diet caused the highest level of serum SGOT and SGPT followed by PKO, RPO, and RBDPO respectively (p < 0.05). The serum SGOT and SGPT were not affected by the AOM treatment. Although the liver color from the AOM group was relatively darker than the non-AOM group, in general the fat sources did not have a significant effect on liver color.

Effects of resveratrol on inhibiting pyroptosis of intestinal cancer cells

Objective: To study the effects of resveratrol (Res) on pyroptosis of colorectal cancer cells . Methods: ①The experiment of dextran sodium sulfate (DSS) induced colon cancer (CRC) in mice: 30 C57BL/6 mice were randomly divided into control group, Azoxymethane (AOM) group, AOM/DSS group, AOM/DSS+Res group and Res group, with 6 mice in each group, the modeling cycle was 70 days in total. Mice in AOM group, AOM/DSS group and AOM/DSS+Res group, at the first day of the first week, were intraperitoneally injected with AOM (10 mg/kg) once, and the ordinary chaw was replaced with high iron feed, and sterile water was given, 1% DSS water was given to AOM/DSS group and AOM/DSS+Res group. The mice in AOM/DSS+Res and Res groups were given resveratrol (50 mg/kg) by oral gavage, When the mold was finished, colon tissue of mice was fixed, embedded and sectionalized. The expressions of NLRP3, Caspase-1 and IL-18 in colon tissues of mice were detected by IHC and Western blot. ②In vitro experiment: HCT 116 cells were given Res (2.4 μg/L) and transfected with miR-31. The Res was divided into 4 groups and labeled with 0 h, 12 h, 24 h and 48 h respectively. The transfected cells were divided into 5 groups: Control group, miR-31 mimic group, miR-31 mimic + Res group, miR-31 inhibitor group, miR-31 inhibitor + Res group. The protein expressions of NLRP3, Caspase-1, GSDMD-N, IL-18 and IL-1β were detected by Western blot. Results: Animal experiments: Compared with control group, the protein expressions of NLRP3, Caspase-1 and IL-18 in AOM/DSS group were increased significantly (P<0.01). The protein expression levels of NLRP3, Caspase-1 and IL-18 in AOM/DSS+Res group were significantly lower than those in AOM/DSS group (P<0.01). Cell experiments: Compared with the control group, the protein expressions of NLRP3 (P<0.01), GSDMD-N (P<0.05) and IL-18 (P< 0.01) in miR-31 mimic group were increased significantly. The protein expressions of NLRP3, GSDMD-N and IL-18 in miR-31 inhibitor group were decreased significantly (P<0.05). Conclusion: Res inhibited the pyroptosis of colorectal cancer cells through pyroptosis.

Fecal DNA Virome Is Associated with the Development of Colorectal Neoplasia in a Murine Model of Colorectal Cancer.

Alteration of the gut virome has been associated with colorectal cancer (CRC); however, when and how the alteration takes place has not been studied. Here, we employ a longitudinal study in mice to characterize the gut virome alteration in azoxymethane (AOM)-induced colorectal neoplasia and identify important viruses associated with tumor growth. The number and size of the tumors increased as the mice aged in the AOM treated group, as compared to the control group. Tumors were first observed in the AOM group at week 12. We observed a significantly lower alpha diversity and shift in viral profile when tumors first appeared. In addition, we identified novel viruses from the genera Brunovirus, Hpunavirus that are positively associated with tumor growth and enriched at a late time point in AOM group, whereas members from Lubbockvirus show a negative correlation with tumor growth. Moreover, network analysis revealed two clusters of viruses in the AOM virome, a group that is positively correlated with tumor growth and another that is negatively correlated with tumor growth, all of which are bacteriophages. Our findings suggest that the gut virome changes along with tumor formation and provides strong evidence of a potential role for bacteriophage in the development of colorectal neoplasia.

Protective effects of rosmarinic acid against azoxymethane-induced colorectal cancer in rats.

Colorectal cancer (CRC) incidence is increasing gradually and has been become one of the most common cancers worldwide. Hence, it is important to discover cheap, naturally occurring compounds to be effective in suppressing the devastating effect of colon-related tumors. Rosmarinic acid (RA), one of the compounds of plant origin, possesses attractive features for use as an agent for cancer prevention and treatment. This study investigated the ability of RA to prevent azoxymethane (AOM)-induced rat colon carcinogenesis by evaluating the effect of RA on tumor formation and circulatory oxidant-antioxidant status. Moreover, plasma levels of adiponectin (APN) monocyte chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6) were detected by enzyme linked immunosorbent assay. The animals were divided into three groups: Control, AOM, and AOM + RA. Rats were fed a modified pellet diet (15.8% peanut oil was added to the standard diet) during the experimental period. Colon cancer was formed by applying 15 mg/kg AOM intraperitoneal once a week for 4 weeks in both the CRC group and AOM + RA group. Besides AOM, AOM + RA group received 5 mg/kg body weight RA orally every day during the study. The results showed that adenocarcinoma rates formed87.5% ofthe AOM group. With treatment of RA, a reduction in the incidence of adenocarcinoma was observed in the AOM + RA group. The plasma MCP-1, IL-6, and TO levels were significantly higher, APN and TAS levels were significantly lower in the AOM group with respect tocontrols. In addition, there was a significant increase in TAS levels in the RA treatment group compared to the AOM group. These findings suggested that RA may be beneficial in preventing AOM-induced colon carcinogenesis formation.

Abstract 2976: A comparative study between the therapeutic efficacy of vitamin D3 and its analogue, paricalcitol, in the treatment of colon cancer in rat

Abstract Background: Vitamin D3 (VD) and its analogue, Paricalcitol (Pcal), have previously shown anti-tumorigenic activities against colorectal cancer (CRC). Hence, this study compared the therapeutic efficacy of both VD types with or without chemotherapy (5-FU) in azoxymethane (AOM)-induced CRC in rat. Methods: seventy male Wistar rats were equally allocated into 7 groups: Control, AOM, VD, Pcal, 5-FU, 5-FU/VD and 5-FU/Pcal. AOM was subcutaneously injected for 2 weeks (15mg/kg/week). The study duration was 25 weeks. VD (500 IU/rat/day; 3 days/week) and Pcal (2.5 μg/kg/day; 3 days/week) were injected subcutaneously from week-15 till the end of the study. 5-FU intraperitoneal injections started in week-21 (12 mg/kg/day; 4 successive days) and continued in week-22 (6 mg/kg/4 doses every other day). The colons were collected for gross and histopathological examination. The expression of VD signalling system, Wnt, β-catenin, NF-κB, COX-2, iNOS, VEGF, smad4, p21 and p27 proteins was measured by immune-histochemistry/fluorescence. The mRNA expression of Wnt, β-catenin, DKK-1 and COX-2 genes was measured by Quantitative RT-PCR. ELISA was used to measure the concentrations of TGF-β1, VEGF and COX-2 proteins in tissues. Results: The AOM and 5-FU groups showed marked inhibitions in the VD receptor and binding protein compared with controls (P &amp;lt; 0.05). VD and Pcal monotherapies significantly increased the expression of the VD molecules and the VD group showed the utmost elevations. Moreover, the tumour numbers and sizes were equally and significantly reduced in the 5-FU, VD and Pcal groups compared with the AOM group (P &amp;lt; 0.05). Interestingly, the 5-FU/VD and 5-FU/Pcal resulted in significantly lower numbers of tumours compared with the remaining study groups and the highest significant reduction was observed in the former group. At the molecular level, the 5-FU/VD markedly inhibited the pro-oncogenic molecules (Wnt, β-catenin, NF-κB, COX-2, iNOS &amp; VEGF) as well as significantly upregulated the anti-tumorigenic molecules (DKK-1, TGF-β1, smad4, p21 and p27) compared with the other study groups (P &amp;lt; 0.05). Conclusions: Vitamin D3 showed a better synergism than Pcal with 5-FU. The enhanced anti-tumorigenic actions of 5-FU/VD involved the modulation of the Wnt/β-catenin and TGF-β1/smad4 pathways, angiogenesis and cell cycle arrest. Further studies are required to elucidate the therapeutic significance of VD in clinical settings Citation Format: Mohammed Abbas Baghdadi, Mohammed Aslam, Jawwad Ahmad, Shakir Idris, Bassem Refaat. A comparative study between the therapeutic efficacy of vitamin D3 and its analogue, paricalcitol, in the treatment of colon cancer in rat [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2976.

Effects of Lactobacillus acidophilus and Bifidobacterium bifidum Probiotics on the Expression of MicroRNAs 135b, 26b, 18a and 155, and Their Involving Genes in Mice Colon Cancer.

A wide range of sources supports that the link between diet and colorectal cancer may be due to an imbalance of the intestinal microflora. In this case, it seems that the probiotics may have a possible molecular mechanism via microRNAs (miRNAs). The present study is aimed to evaluate the effects of Lactobacillus acidophilus and Bifidobacterium bifidum probiotics on the expression of miRNAs 135b, 26b, 18a, and 155 and their target genes, including APC, PTEN, KRAS, and PU.1 in mouse azoxymethane (AOM)-induced colon cancer. Thirty-eight male BALB/c mice were randomly divided into four groups: the control, AOM, Lactobacillus acidophilus, and Bifidobacterium bifidum to deliberate the effects of the probiotics on the miRNAs and their target genes. Except for the control group, the rest groups were weekly given AOM (15mg/kg, s.c) in three consecutive weeks to induce mouse colon cancer. The animals were given 1.5g powders of L. acidophilus (1 × 109cfu/g) and B. bifidum (1 × 109cfu/g) in 30cc drinking water in the related groups for 5months. At the end of the study, the animals were sacrificed and their blood and colon samples were removed for the molecular analyses. The results showed that the expression of the miR-135b, miR-155, and KRAS was increased in the AOM group compared to the control group in both the plasma and the colon tissue samples, and the consumption of the probiotics decreased their expression. Moreover, the miR-26b, miR-18a, APC, PU.1, and PTEN expressions were decreased in the AOM group compared to the control group and the consumption of the probiotics increased their expressions. It seems that Lactobacillus acidophilus and Bifidobacterium bifidum though increasing the expression of the tumor suppressor miRNAs and their target genes and decreasing the oncogenes can improve colon cancer treatment.

The protective effects of fermented kefir milk on azoxymethane-induced aberrant crypt formation in mice colon.

Kefir is a probiotic fermented milk product produced from grains with a complex composition of bacteria and yeasts that live in a symbiotic association. Anti-proliferative, anti-inflammatory, and anti-mutagenic effects are some of the health beneficial properties of kefir grains. The present study was conducted to evaluate whether regular consumption of kefir milk would be capable of preventing the development of pre-neoplastic lesions induced by azoxymethane (AOM). Aberrant crypt foci were induced in BALB-c mice via 2 subcutaneous injections of azoxymethane (15 mg/kg) and kefir was administered by daily gavage for 8 weeks (5 ml/kg). Additionally, bacterial growth was monitored in pasteurized and ultra-high temperature (UHT) treated milk to compare different fermentation conditions. Our results showed that UHT milk presented better growth of Lactobacillus acidophilus colonies. The aberrant crypt foci were attenuated by approximately 43% (height) and 20% (width) in the kefir group compared to AOM group, suggesting that kefir treatment may contribute to prevent and control the growth of intestinal neoplastic cells.

The Effect of Carica pubescens Leaf Extract on Haematological Profile and Histopathological Appearance of Azoxymethane-Induced Colon Cancer Rats

Event Abstract Back to Event Effects of Carica pubescens leaf extract on haematological profile and histopathological appearance of azoxymethane-induced colon cancer rats Ainun R. Gumay1*, Saekhol Bakri1, Eka M. Sugeng1, Maharani S. Yudina1, Faradilla N. Bleizensky1, Muflihatul Muniroh1, Yosef Purwoko1 and Hardian Hardian1 1 Diponegoro University, Indonesia ABSTRACT Background Neuroinflammatory and apoptosis mechanisms play an important role in the pathogenesis of colorectal cancer. Carica pubescens (CP) has potential anti-inflammatory, antioxidant and anticancer effects. This study aims to determine the effect of CP leaf extract on haematological profile and histopathological appearance of azoxymethane (AOM)-induced colon cancer rats. Methods Sprague Dawley rats (25 male,aged 5-7weeks) were divided into 5 groups. The AOM groups were induced intraperitoneally by injection of AOM (10 mg/kgBW) once per week for 2 weeks. CP-100, CP-200, and CP-400 groups were induced by AOM and orally administered with 100, 200, and 400 mg/kgBW CP leaf extract once daily for 2 weeks. The normal control (NC) group was given saline. The haematological profile that was examined includes white blood cell (WBC), lymphocyte, and neutrophil count. The histopathological appearance shows the number of inflammatory cells (macrophages, lymphocytes, histiocyte, and polimorphonuclear cells). One-way Anova and post hoc LSD were used for statistical analysis. Results The WBC count of CP-100 (4380±715.5/μL) was significantly lower than AOM (7000±2065.2/μL; p=0.002). The lymphocyte count of CP-100 (3260±746.9/μL) was also significantly lower than AOM (5460±1647.1/μL; p=0.001). The neutrophil counts in CP-100 (940±554.9/μL; p=0.005), CP-200 (1220±342.1/μL;p=0.001), and CP-400 (1240±680.4/μL; p=0.008) were significantly lower than the AOM group (2040±270.2/μL). The mean of inflammatory cells in histopathological appearance of AOM group (1008.6±21.45 cells) was significantly higher than N-C group (412.6±14.42 cells), CP-100 (843.4±13.72 cells), CP-200 (701.2±25.41 cells), and CP-400 (642.0±30.00 cells) with p<0.001. Conclusion Carica pubescens leaf extract seem to reduce the number of inflammatory cells, WBC, lymphocytes, and neutrophil counts in azoxymethane-induced colon cancer rats. Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Acknowledgements We thank the Faculty of Medicine Diponegoro University, Indonesia for funding of the project (Grant Num 596/UN7.5.4/HK/PP/2018) Keywords: Carica pubescens, Azoxymethane (AOM), Colon cancer (CC), Antiinflamation, Chemoprevention Conference: International Conference on Drug Discovery and Translational Medicine 2018 (ICDDTM '18) “Seizing Opportunities and Addressing Challenges of Precision Medicine”, Putrajaya, Malaysia, 3 Dec - 5 Feb, 2019. Presentation Type: Oral Presentation Topic: Cancer Citation: Gumay AR, Bakri S, Sugeng EM, Yudina MS, Bleizensky FN, Muniroh M, Purwoko Y and Hardian H (2019). Effects of Carica pubescens leaf extract on haematological profile and histopathological appearance of azoxymethane-induced colon cancer rats. Front. Pharmacol. Conference Abstract: International Conference on Drug Discovery and Translational Medicine 2018 (ICDDTM '18) “Seizing Opportunities and Addressing Challenges of Precision Medicine”. doi: 10.3389/conf.fphar.2018.63.00089 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 29 Nov 2018; Published Online: 17 Jan 2019. * Correspondence: Mrs. Ainun R Gumay, Diponegoro University, Semarang, Indonesia, ainungumay@fk.undip.ac.id Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Ainun R Gumay Saekhol Bakri Eka M Sugeng Maharani S Yudina Faradilla N Bleizensky Muflihatul Muniroh Yosef Purwoko Hardian Hardian Google Ainun R Gumay Saekhol Bakri Eka M Sugeng Maharani S Yudina Faradilla N Bleizensky Muflihatul Muniroh Yosef Purwoko Hardian Hardian Google Scholar Ainun R Gumay Saekhol Bakri Eka M Sugeng Maharani S Yudina Faradilla N Bleizensky Muflihatul Muniroh Yosef Purwoko Hardian Hardian PubMed Ainun R Gumay Saekhol Bakri Eka M Sugeng Maharani S Yudina Faradilla N Bleizensky Muflihatul Muniroh Yosef Purwoko Hardian Hardian Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Chemopreventive effects of Ku-jin tea against AOM-induced precancerous colorectal lesions in rats and metabolomic analysis

Ku-jin tea (KJT) is a health beverage prepared from the leaves of the plant Acer tataricum subsp. ginnala that has been consumed in some regions of China for thousands of years. KJT contains high levels of anti-inflammatory and antioxidative compounds such as ginnalins, but little is known about the chemopreventive effect of KJT on colon cancer. In this study, we investigated the preventive effects of KJT on colon carcinogenesis using the azoxymethane (AOM)-induced precancerous colorectal lesion model in rats. The results showed that the number of aberrant crypts, aberrant crypt foci (ACF) and crypts/focus in rats of the KJT + AOM group were significantly decreased compared with rats of the AOM group (p < 0.01). Further exploration of the prevention mechanism of KJT by UPLC-QTOF/MS-based urinary metabolomics showed that 5 metabolic pathways were modulated, including purine metabolism and amino acid metabolism, in the group with KJT. In addition, the levels of the immunomodulatory cytokines IL-1α and IL-10 were significantly decreased, and the levels of IL-2 in the serum of AOM rats increased after KJT treatment. Our present data suggest that KJT can inhibit AOM-induced colonic ACF formation and might be a useful chemopreventive agent against colorectal carcinogenesis.

Preventive effects of Resveratrol against azoxymethane-induced testis injury in rats.

To evaluate the protective effects of Resveratrol (RES) on azoxymethane (AOM)-induced testicular damage using histopathology and biochemical analyses, 28 male rats were randomly divided into four groups. Groups were control, RES, AOM and ARES. At the end of the 7weeks, following routine tissue processing procedure, testis sections were stained with haematoxylin-eosin and Masson's trichrome. The blood samples were taken for biochemical analysis of testosterone, total oxidative stress, total antioxidant status and oxidative stress index. Degenerative changes in the seminiferous tubules such as atrophy, loss in the number of germ cells and arrested spermatogenic cell, and increase in the connective tissue of the tunica albuginea in the groups with AOM treatment were found. RES treatment (ARES) reduced the number of affected seminiferous tubules significantly (p<.05) compared to AOM alone. The testosterone levels in AOM group were significantly lower than in the control group (p<.05). The total oxidative stress levels were significantly higher in AOM group compared to control group (p<.05). The total antioxidant status levels in ARES group were significantly higher than in the AOM group (p<.05). This study results suggest that an antioxidant like Resveratrol may be useful for decreasing the harmful effects of azoxymethane.

Brown seaweeds protect against azoxymethane-induced hepatic repercussions through up-regulation of peroxisome proliferator-activated receptor gamma and attenuation of oxidative stress

Context: Seaweeds of the genera Turbinaria and Padina have long been used as food and in traditional medicine for treating several diseases.Objective: The current study determines the protective efficacy of the brown seaweeds Turbinaria ornata (Turner) J. Agardh (Sargassaceae) and Padina pavonia (Linnaeus) J.V. Lamouroux (Dictyotaceae) against liver injury induced by azoxymethane (AOM).Materials and methods: Male Swiss mice received 10 mg/kg AOM once a week for two consecutive weeks and then 100 mg/kg daily dose of either T. ornata or P. pavonia ethanolic extract. Thirteen weeks after the first AOM administration and 24 h after the last treatment, overnight fasted mice were sacrificed and samples collected.Results: Compared with the AOM group, both T. ornata and P. pavonia significantly decreased the activity of aminotransferases and the concentration of bilirubin while increased albumin levels in the serum. The antioxidative effect of both extracts was observed from the increased activity of superoxide dismutase and glutathione peroxidase activities in the liver, both of which were decreased by AOM. Moreover, the levels of malondialdehyde and nitric oxide were reduced, and histological findings also confirmed the antihepatotoxic activity. In addition, treatment with T. ornata and P. pavonia significantly increased PPARγ and decreased NF-κB expression in the liver of AOM-administered mice.Discussion and conclusion: Our findings indicate that the protective function of T. ornata and P. pavonia on AOM-induced liver injury may be possibly exerted by multiple pathways including abolishment of inflammation and oxidative damage, and activation of PPARγ.

Abstract 907: Chemopreventive effects of a non-camellia tea against azoxymethane -induced precancerous colorectal lesions in male rats

Abstract Tea (the leaves from Camellia plants) is one of the most popular beverages in the world; substantial evidence supports its success in the prevention of carcinogenesis and other chronic diseases in animal models. Plants other than Camellia have been widely used as tea as well in various cultures. We have systematically collected and identified over 30 species of frequently used Non-Camellia teas, which have been consumed at least 300 years in China or other nations. Recently, non-Camellia teas have attracted much attentions due to high potentials of preventing and treating metabolic and inflammatory diseases, as well as cancers. Epidemiologic studies report that tea consumption significantly reduces risk of colorectal cancer (CRC). We here reported in vivo preventive effects of the Ku-jin-cha (KJC) tea for CRC. KJC tea is a kind of “Non-Camellia tea” prepared from leaves of the plant Acer ginnala and popularly used in some Chinese regions for thousands of years. KJC contains high levels of anti-inflammatory and anti-oxidative compounds such as ginnalins. However, the potential chemopreventive effect of KJC remains unknown. Here, we investigated modulatory effects of KJC on colon carcinogenesis using the rat azoxymethane (AOM) induced precancerous colorectal lesion model. Wistar rats (male, 4∼5 weeks old) were randomly divided into 4 groups (Control, KJC, AOM and AOM+KJC, 10 rats/group) and were fed either normal water or experimental water containing KJC for 12 weeks. After one week of water or KJC drinking, all rats except the Control Group were injected with 15 mg/kg AOM (s.c., once weekly for 3 weeks). At 16∼17 weeks of age, urine and sera were collected for metabolomic analysis using UPLC-QTOF/MS (ultra-HPLC coupled with time-of-flight mass spectrometry). Rats were sacrificed at the end of KJC administration, and colorectums collected for evaluation of aberrant crypt (AC), aberrant crypt foci (ACF) and Crypts/focus. Spleens, thymuses and bloods were also taken for immunological analysis. Our results show that the numbers of AC, ACF and Crypts/focus in rats of the KJC+AOM Group were significantly decreased, versus rats of the AOM Group (p &amp;lt; 0.01). No ACF was found in rats without AOM injection. In addition, significant metabolic alterations were observed in the groups with KJC. Although KJC administration did not cause marked weight changes of spleens and thymuses, the effect of KJC on levels of inflammatory cytokines and mediators were also examined. Our results indicated that KJC significantly inhibited AOM-induced colonic ACF formation, suggesting that KJC might have a chemopreventive effect against colon carcinogenesis at least in the initiation stage of CRC. Furthermore, KJC may exert the chemopreventive effects on ACF formation by changing metabolic pathways and modulating immunity and inflammatory responses. This warrants further studies on the cellular and molecular mechanisms of KJC tea. Citation Format: Chunnian He, Wu Bi, L. Harris Zhang, Li Cao, Pei-Gen Xiao. Chemopreventive effects of a non-camellia tea against azoxymethane -induced precancerous colorectal lesions in male rats. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 907. doi:10.1158/1538-7445.AM2015-907

Grape juice concentrate (G8000™) modulates apoptosis but not oxidative stress following rat colon carcinogenesis induced by azoxymethane

Objectives: The aim of this study was to evaluate if grape juice concentrate is able to protect against experimental colon carcinogenesis.Material and methods: For this purpose, a total of 35 male Wistar rats were randomly distributed into seven groups: G1: SHAM animals receiving only saline; G2: animals receiving 15 mg/kg azoxymethane (AOM); G3: animals receiving 1% grape juice concentrate 2 weeks before the administration of AOM; G4: animals receiving 2% grape juice concentrate 2 weeks before the administration of AOM; G5: animals receiving 1% grape juice concentrate 4 weeks after the last administration of AOM; G6: animals receiving 2% grape juice concentrate 4 weeks after the last administration of AOM; G7: animals receiving only 2% grape juice concentrate.Results: The group that received 2% grape juice concentrate before induction with AOM showed the decreased expression of Bcl-2 compared to those animals that were induced by AOM (positive control). Regarding Bax, animals that received grape juice at 2% decreased Bax immunoexpression when compared to AOM group. Furthermore, animals that intake grape juice at 1% after induced by AOM decreased Bax immunoexpression as well. 8-OHdGLI did not show significant statistically differences (p > 0.05) among groups.Conclusion: In summary, our results demonstrate that grape juice is able to modulate rat colon carcinogenesis as a result of induction of apoptosis.

Chemopreventive efficacy of Andrographis paniculata on azoxymethane-induced aberrant colon crypt foci in vivo.

Andrographis paniculata is a grass-shaped medicinal herb, traditionally used in Southeast Asia. The aim of this study was to evaluate the chemoprotective effects of A. paniculata on colorectal cancer. A. paniculata ethanol extract was tested on azoxymethane (AOM)-induced aberrant crypt foci (ACF) in vivo and in vitro. A. paniculata treated groups showed a significant reduction in the number of ACF of the treated rats. Microscopically, ACF showed remarkably elongated and stratified cells, and depletion of the submucosal glands of AOM group compared to the treated groups. Histologically, staining showed slightly elevated masses above the surrounding mucosa with oval or slit-like orifices. Immunohistochemically, expression of proliferating cell nuclear antigen (PCNA) and β-catenin protein were down-regulated in the A. paniculata treated groups compared to the AOM group. When colon tissue was homogenized, malondialdehyde (MDA) and nitric oxide (NO) levels were significantly decreased, whereas superoxide dismutase (SOD) activity was increased in the treated groups compared to the AOM group. A. paniculata ethanol extract showed antioxidant and free radical scavenging activity, as elucidated by the measure of oxidative stress markers. Further, the active fractions were assessed against cell lines of CCD841 and HT29 colon cancer cells.

Antioxidant and Anti-inflammatory Effects of Yam (Dioscorea batatas Decne.) on Azoxymethane-induced Colonic Aberrant Crypt Foci in F344 Rats.

Yam (Dioscorea batatas Decne.) has long been used as a health food and oriental folk medicine because of its nutritional fortification, tonic, anti-diarrheal, anti-inflammatory, antitussive, and expectorant effects. Reactive oxygen species (ROS), which are known to be implicated in a range of diseases, may be important progenitors of carcinogenesis. The aim of this study was to investigate the modulatory effect of yam on antioxidant status and inflammatory conditions during azoxymethane (AOM)-induced colon carcinogenesis in male F344 rats. We measured the formation of aberrant crypt foci (ACF), hemolysate antioxidant enzyme activities, colonic mucosal antioxidant enzyme gene expression, and colonic mucosal inflammatory mediator gene expression. The feeding of yam prior to carcinogenesis significantly inhibited AOM-induced colonic ACF formation. In yam-administered rats, erythrocyte levels of glutathione, glutathione peroxidase (GPx), and catalase were increased and colonic mucosal gene expression of Cu/Zn-superoxide dismutase (SOD), Mn-SOD, and GPx were up-regulated compared to the AOM group. Colonic mucosal gene expression of inflammatory mediators (i.e., nuclear factor kappaB, inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor alpha, and interleukin-1beta) was suppressed by the yam-supplemented diet. These results suggest that yam could be very useful for the prevention of colon cancer, as they enhance the antioxidant defense system and modulate inflammatory mediators.

A non-digestible fraction of the common bean (Phaseolus vulgaris L.) induces cell cycle arrest and apoptosis during early carcinogenesis.

We have previously demonstrated that the non-digestible fraction (NDF) from common cooked beans (P. vulgaris L., cv Negro 8025) inhibits azoxymethane (AOM)-induced colon cancer and influences the expression of genes involved in the induction of apoptosis and cell cycle arrest through the action of butyrate. The objective of this study was to identify cell cycle alterations and morphological changes induced by treatment with AOM and to examine the formation of colonic aberrant crypt foci (ACF) in male Sprague Dawley rats fed with these beans. Rats were fed control diets upon arrival and were randomly placed into four groups after one week of acclimatization: control, NDF (intragastric administration), NDF + AOM and AOM. Rats treated with NDF + AOM exhibited a significantly lower number of total colonic ACF with a notable increase in the number of cells present in the G1 phase (83.14%); a decreased proliferation index was observed in the NDF + AOM group when compared to AOM group. NDF + AOM also displayed a higher number of apoptotic cells compared to AOM group. NDF of cooked common beans inhibited colon carcinogenesis at an early stage by inducing cell cycle arrest of colon cells and morphological changes linked to apoptosis, thus confirming previous results obtained with gene expression studies.

Flaxseed (Linum usitatissimum L.) and Its Total Non-digestible Fraction Influence the Expression of Genes Involved in Azoxymethane-induced Colon Cancer in Rats

The influence of flaxseed (Linum usitatissimum L.) and its total non-digestible fraction (TNDF) on the expression of genes involved in azoxymethane (AOM)-induced colon cancer in Sprague Dawley rats was analyzed. The dose used in the animal model was two tablespoons of flaxseed per day, which is the dose recommended for humans. Flaxseed significantly decreased the crypt multiplicity (10.50 ± 3.5) compared with the AOM treatment (34.00 ± 11.0), which suggests that flaxseed exhibits a preventive effect against colon cancer. Both treatments (flaxseed and TNDF) influence the overexpression of genes involved in cell cycle arrest and mitochondrial apoptosis: p53, p21, bcl-2, bax and caspase-3. Flaxseed induced the expression of p53 and p21, whereas TNDF triggered the p21-independent expression of p53. This finding suggests that both of these treatments induced cell cycle arrest. In addition, TNDF induced mitochondrial apoptosis because the TNDF + AOM group exhibited the expression of caspase-3, decreased bcl-2 expression and increased bax expression. These results suggest that the expression of the analyzed genes is associated with the presence of dietary antioxidants linked to the cell wall of flaxseed.

Preventive Effects of Resveratrol against Azoxymethane Induced Damage in Rat Liver

In recent years, due to modern lifestyles and exposure to chemical carcinogens, cancer cases are steadily increasing. From this standpoint, azoxymethane (AOM), a chemical carcinogen which causes de novo liver damage, and resveratrol, which is an antioxidant found in foods and protects against oxidative stress damage, are of interest. We here aimed to evaluate whether resveratrol could protect the liver tissues from the effects of AOM. The study was conducted in 4 groups, each consisting of seven rats, the first receiving only AOM (2 times per week, 5 mg/kg), group 2 AOM and resveratrol (2 times a week, 20 mg/kg), group 3 assessed only as a control and group 4 administered only resveratrol. At the end of the seventh week, the rats were sacrificed. Rat liver MDA, NO, GSH levels were analyzed biochemically, as well as the tissues being evaluated histopathologically. MDA and NO increased in AOM group as signs of increased oxidative stress. The group concomitantly administered resveratrol was been found to be significantly decreased in MDA and NO levels and increased in GSH activity. However, there were no significant findings on histopathological evaluation. In the light of these results, resveratrol appears to exert protective effect on oxidative stress in the liver tissue due to deleterious effects of chemical carcinogens.