Effects of 3,3’-Diindolylmethane on Rat Kidney Tissue

Abstract

Aim: 3,3’-diindolylmethane (DIM) is an important digestive product of indole-3 carbinol (I3C) obtained from the Brassica family (broccoli, cauliflower, cabbage, etc.) of vegetables. 3,3’-diindolylmethane is a substrate with potent immune modulatory activity and antitumor, antiviral, and anti-angiogenic effects. This study aimed to evaluate the effects of DIM on rat kidney tissue using histopathologic methods. Material and Method: In the study, 36 males, 16-week-old, and 220–260 gr Wistar albino adult rats were used. Rats were divided into four equal groups: The control group received only corn oil by oral gavage. The other experimental groups received three different doses of DIM dissolved in corn oil, 10 mg/kg DIM (DIM 10 group), 50 mg/kg DIM (DIM 50 group), and 100 mg/kg DIM (DIM- 100 group, were administered via oral gavage. Oral gavages were applied to experimental groups for 53 days. At the end of the experiment, all rats were euthanized, and the kidney tissues were dissected. For histopathological examination, the kidney tissue samples were stained with hematoxylin-eosin and Masson–trichrome. Results: Our investigation revealed that the use of DIM at different doses for 53 days caused dose-dependent histopathological changes, including apoptotic to necrotic changes, interstitial inflammation to fibrotic connective tissue changes, and cast formations starting from the Henle loops and spreading to the renal tubules. Conclusion: These histopathological changes could have occurred due to a DIM-mediated increase in reactive oxygen species (ROS). Further biochemical, molecular, and ultrastructural studies are needed to clarify these findings.

Keywords

• 3
• 3’-diindolylmethane
• indole-3 carbinol
• kidney; reactive oxygen species (ROS)

References

1. 1. Okuyaz S, Tamer A. Mersin Üniversitesi Tıp Fakültesi Lokman Hekim Tıp Tarihi ve Folklorik Tıp Dergisi üzerine bir araştırma:2011’den günümüze. Mersin Üniversitesi Tıp Fakültesi Lokman Hekim Tıp Tarihi ve Folklorik Tıp Dergisi, 13(2):445–452, 2023.
2. 2. Rogan EG. The natural chemopreventive compound indole-3-carbinol: state of the science. in vivo, 20(2):221–228, 2006.
3. 3. Banerjee S, Wang Z, Kong D, Sarkar FH. 3, 3′-Diindolylmethane enhances chemosensitivity of multiple chemotherapeutic agents in pancreatic cancer. Cancer research, 69(13):5592–5600, 2009.
4. 4. Kunimasa K, Kobayashi T, Kaji K, Ohta T. Antiangiogenic effects of indole-3-carbinol and 3, 3′-diindolylmethane are associated with their differential regulation of ERK1/2 and Akt in tube-forming HUVEC. The Journal of nutrition, 140(1):1–6, 2010.
5. 5. Chang X, Tou JC, Hong C, Kim H-A, Riby JE, Firestone GL, et al. 3, 3′-Diindolylmethane inhibits angiogenesis and the growth of transplantable human breast carcinoma in athymic mice.Carcinogenesis, 26(4):771–778, 2005.
6. 6. Xue L, Pestka JJ, Li M, Firestone GL, Bjeldanes LF. 3,3′-Diindolylmethane stimulates murine immune function in vitro and in vivo. The Journal of nutritional biochemistry, 19(5):336–344, 2008. 7. Wang TT, Schoene NW, Milner JA, Kim YS. Broccoli‐derived phytochemicals indole‐3‐carbinol and 3, 3′‐diindolylmethane exerts concentration‐dependent pleiotropic effects on prostate cancer cells: Comparison with other cancer preventive phytochemicals. Molecular carcinogenesis, 51(3):244–256, 2012.
7. 8. Banerjee S, Kong D, Wang Z, Bao B, Hillman GG, Sarkar FH. Attenuation of multi-targeted proliferation-linked signaling by 3, 3′-diindolylmethane (DIM): from bench to clinic. Mutation Research/Reviews in Mutation Research, 728(1–2):47–66, 2011.
8. 9. Kim B-G, Kim J-W, Kim S-M, Go R-E, Hwang K-A, Choi K-C. 3, 3′-Diindolylmethane Suppressed Cyprodinil-Induced Epithelial-Mesenchymal Transition and Metastatic-Related Behaviors of Human Endometrial Ishikawa Cells via an Estrogen Receptor-Dependent Pathway. International Journal of Molecular Sciences, 19(1):189, 2018.

9. 10. Amare DE. Anti-cancer and other biological effects of a dietary compound 3, 3′-diindolylmethane supplementation: a systematic review of human clinical trials. Nutrition and Dietary Supplements:123–137, 2020.
10. 11. Firestone GL, Bjeldanes LF. Indole-3-carbinol and 3–3′-diindolylmethane antiproliferative signaling pathways control cell-cycle gene transcription in human breast cancer cells by regulating promoter-Sp1 transcription factor interactions. The Journal of nutrition, 133(7): c-2455S, 2003.
11. 12. Savino III JA, Evans JF, Rabinowitz D, Auborn KJ, Carter TH. Multiple, disparate roles for calcium signaling in apoptosis of human prostate and cervical cancer cells exposed to diindolylmethane. Molecular cancer therapeutics, 5(3):556– 563, 2006.
12. 13. Kim EJ, Park SY, Shin H-K, Kwon DY, Surh Y-J, Park JHY. Activation of caspase-8 contributes to 3, 3′-Diindolylmethaneinduced apoptosis in colon cancer cells. The Journal of nutrition, 137(1):31–36, 2007.
13. 14. Abdelrahim M, Newman K, Vanderlaag K, Samudio I, Safe S. 3, 3′-diindolylmethane (DIM) and its derivatives induce apoptosis in pancreatic cancer cells through endoplasmic reticulum stressdependent upregulation of DR5. Carcinogenesis, 27(4):717–728, 2006.
14. 15. Acharya A, Das I, Singh S, Saha T. Chemopreventive properties of indole-3-carbinol, diindolylmethane and other constituents of cardamom against carcinogenesis. Recent patents on food, nutrition & agriculture, 2(2):166–177, 2010.
15. 16. Aksu E, Akman O, Ömür A, Karakuş E, Can I, Kandemir F, et al. 3, 3 diindolylmethane leads to apoptosis, decreases sperm quality, affects blood estradiol 17 β and testosterone, oestrogen (α and β) and androgen receptor levels in the reproductive system in male rats. Andrologia, 48(10):1155–1165, 2016.
16. 17. Rouse M, Rao R, Nagarkatti M, Nagarkatti PS. 3,3′-diindolylmethane ameliorates experimental autoimmune encephalomyelitis by promoting cell cycle arrest and apoptosis in activated T cells through microRNA signaling pathways. Journal of Pharmacology and Experimental Therapeutics, 350(2):341–352, 2014.
17. 18. Tomar S, Nagarkatti M, Nagarkatti P. 3, 3′‐Diindolylmethane attenuates LPS‐mediated acute liver failure by regulating miRNAs to target IRAK4 and suppress Toll‐like receptor signalling. British journal of pharmacology, 172(8):2133–2147, 2015.
18. 19. Lee BD, Yoo J-M, Baek SY, Li FY, Sok D-E, et al. 3, 3′-Diindolylmethane promotes BDNF and antioxidant enzyme formation via TrkB/Akt pathway activation for neuroprotection against oxidative stress-induced apoptosis in hippocampal neuronal cells. Antioxidants, 9(1):3, 2019.
19. 20. Lerner A, Grafi-Cohen M, Napso T, Azzam N, Fares F. The indolic diet-derivative, 3, 3′-diindolylmethane, induced apoptosis in human colon cancer cells through upregulation of NDRG1. BioMed Research International, 2012, 2012.
20. 21. Bui PV, Moualla M, Upson DJ. A possible association of diindolylmethane with pulmonary embolism and deep venous thrombosis. Case Reports in Medicine, 2016, 2016.
21. 22. Gong Y, Sohn H, Xue L, Firestone GL, Bjeldanes LF. 3,3′-Diindolylmethane is a novel mitochondrial H+-ATP synthase inhibitor that can induce p21Cip1/Waf1 expression by induction of oxidative stress in human breast cancer cells. Cancer research, 66(9):4880–4887, 2006.
22. 23. Ye Y, Li X, Feng G, Ma Y, Ye F, Shen H, et al. 3, 3′-Diindolylmethane induces ferroptosis by BAP1-IP3R axis in BGC-823 gastric cancer cells. Anti-Cancer Drugs, 33(4):362–370, 2022.
23. 24. Reyes-Hernández OD, Figueroa-González G, Quintas- Granados LI, Gutiérrez-Ruíz SC, Hernández-Parra H, Romero-Montero A, et al. 3, 3′-Diindolylmethane and indole-3-carbinol: potential therapeutic molecules for cancer chemoprevention and treatment via regulating cellular signaling pathways. Cancer Cell International, 23(1):180, 2023.
24. 25. Syed RU, Moni SS, Break MKB, Khojali WM, Jafar M, Alshammari MD, et al. A multi-faceted vegetable for health: An in-depth review of its nutritional attributes, antimicrobial abilities, and anti-inflammatory properties. Antibiotics, 12(7):1157, 2023.
25. 26. Garcia-Ibañez P, Núñez-Sánchez MA, Oliva-Bolarín A, Martínez-Sánchez MA, Ramos-Molina B, Ruiz-Alcaraz AJ, et al. Anti-inflammatory potential of digested Brassica sprout extracts in human macrophage-like HL-60 cells. Food & Function, 14(1):112–121, 2023.
26. 27. Amarakoon D, Lee W-J, Tamia G, Lee S-H. Indole-3-Carbinol: Occurrence, health-beneficial properties, and cellular/molecular mechanisms. Annual Review of Food Science and Technology, 14:347–366, 2023.
27. 28. He J, Huang T, Zhao L. 3, 3’‑Diindolylmethane mitigates lipopolysaccharide‑induced acute kidney injury in mice by inhibiting NOX‑mediated oxidative stress and the apoptosis of renal tubular epithelial cells. Molecular Medicine Reports, 19(6):5115–5122, 2019.
28. 29. Xia Z-E, Xi J-L, Shi L. 3, 3′-Diindolylmethane ameliorates renal fibrosis through the inhibition of renal fibroblast activation in vivo and in vitro. Renal failure, 40(1):447–454, 2018.
29. 30. Choi K-M, Yoo H-S. Amelioration of hyperglycemia-induced nephropathy by 3, 3′-diindolylmethane in diabetic mice. Molecules, 24(24):4474, 2019.
30. 31. Jayakumar P, Pugalendi KV, Sankaran M. Attenuation of hyperglycemia-mediated oxidative stress by indole-3-carbinol and its metabolite 3, 3′-diindolylmethane in C57BL/6J mice. Journal of physiology and biochemistry, 70:525–534, 2014.
31. 32. Zhu J, Li Y, Guan C, Chen Z. Anti-proliferative and proapoptotic effects of 3, 3’-diindolylmethane in human cervical cancer cells. Oncology reports, 28(3):1063–1068, 2012.
32. 33. Roh YS, Cho A, Islam MR, Cho S-D, Kim J, Kim JH, et al. 3,3′-Diindolylmethane induces immunotoxicity via splenocyte apoptosis in neonatal mice. Toxicology letters, 206(2):218–228, 2011.
33. 34. Goldberg AA, Titorenko VI, Beach A, Abdelbaqi K,Safe S, Sanderson JT. Ring-substituted analogs of 3,3′-diindolylmethane (DIM) induce apoptosis and necrosis in androgen-dependent and-independent prostate cancer cells. Investigational new drugs, 32:25–36, 2014.
34. 35. Simon H-U, Haj-Yehia A, Levi-Schaffer F. Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis, 5:415–418, 2000.
35. 36. Gach K, Długosz A, Janecka A. The role of oxidative stress in anticancer activity of sesquiterpene lactones. Naunyn- Schmiedeberg’s archives of pharmacology, 388:477–486, 2015.
36. 37. Higuchi M, Honda T, Proske RJ, Yeh ET. Regulation of reactive oxygen species-induced apoptosis and necrosis by caspase 3-like proteases. Oncogene, 17(21):2753–2760, 1998.
37. 38. Morgan MJ, Kim Y-S, Liu Z-g. TNFα and reactive oxygen species in necrotic cell death. Cell research, 18(3):343–349, 2008.
38. 39. Leibelt DA, Hedstrom OR, Fischer KA, Pereira CB, Williams DE. Evaluation of chronic dietary exposure to indole-3-carbinol and absorption-enhanced 3, 3′-diindolylmethane in spraguedawley rats. Toxicological Sciences, 74(1):10–21, 2003.
39. 40. Tripathi P, Hildeman D. Sensitization of T cells to apoptosis—a role for ROS? Apoptosis, 9:515–523, 2004.
40. 41. Martínez-Klimova E, Aparicio-Trejo OE, Tapia E, Pedraza- Chaverri J. Unilateral ureteral obstruction as a model to investigate fibrosis-attenuating treatments. Biomolecules, 9(4):141, 2019.