The THE ROLE OF PLATELET RICH PLASMA AND QUERCETIN IN ALLEVIATING DIMETHYLNITROSAMINE-INDUCED ACUTE SPLEEN INJURY THROUGH REGULATING OXIDATIVE STRESS, INFLAMMATION AND APOPTOSIS
Objective: Excessive oxidative stress is implicated in spleen injury. Platelet-rich plasma (PRP) and quercetin (QUR) have been shown to protect cells against oxidative stress. This study was designed to investigate their effect on dimethyl nitrosamine (DMN) induced spleen injury in male rats.
Methods: Forty male Wistar rats were divided into four groups; Group (1): Negative control group (Con), Group (2): DMN group, DMN was given intraperitonealy at a dose of 4 mg/kg b. wt/day for four weeks for sub-chronic injury of spleen tissue, Group (3): DMN+PRP, rats were injected intraperitonealy with DMN at a dose of 4 mg/kg b. wt/day for four weeks then treated i. v. by single dose 50 μL of PRP, then left for a period of four weeks without any treatments, Group(4): DMN+QUR, rats received intraperitonealy DMN at a dose of 4 mg/kg b. wt/day for four weeks, then treated with quercetin orally at a dose of 50 mg/kg b. wt. in aqueous suspension daily using an intragastric tube for four weeks.
Results: DMN inoculation resulted in significant elevations of oxidative stress, as evidenced by the increased malondialdehyde, hydrogen peroxide and xanthine oxidase levels associated with a significant decrease in Superoxide dismutase and catalase activities in the spleen tissue as compared to the normal control group. Moreover, DMN caused an up-regulation in the values of the splenic C-reactive protein (CRP), interleuckin-6 (IL-6), nuclear factor kappa B (NF-κB), leukotriene-C4 (LT-C4), P53 and Fas levels with a significant decline in anti-apoptotic protein B-cell lymphoma 2 level as compared to the normal control group. PRP and QUR significantly attenuated the DMN-evoked spleen oxidative stress and modulated the activities of antioxidant enzymes as compared to DMN group. In addition, treatment of DMN group with PRP or QUR resulted in an improvement in CRP, IL-6, NF-κB, LT-C4, P53 and Fas levels as compared to DMN group. Caspase-3 expression was positive in DMN group while no difference was present in control, PRP and Quercetin groups. However, the VEGF immunopositive reaction was found in DMN, PRP and Quercetin groups compared to control group. Histopathological results showed degeneration, haemorrhage, inflammatory cells and necrotoic areas in splenic tissue from DMN group compared to the treated groups where signs of recovery were observed in the whole splenic tissue.
Conclusion: These data suggest that PRP and QUR protect rat spleen from DMN-induced oxidative stress, probably via their antioxidant activity, anti-inflammatory and anti-apoptotic effects. So, PRP and QUR are promising pharmacological agents for preventing the potential spleen injury of DMN following occupational or environmental exposures.
2. El-Shenawy NS, Hamza RZ, Ismail HAA, Khaled HE. Efficacy of ?–lipoic acid against oxidative stress and histopathological changes of dimethylnitrosamine in liver male mice. Am J Biochem Mol Biol 2016;6:102-12.
3. Hamza RZ, Ismail HAA, El-Shenawy NS. Oxidative stress, histopathological and electron microscopically alternation induced by dimethylnitrosamine in renal male mice and the protective effect of ?–lipoic acid. J Basic Clin Physiol Pharmacol 2017;28:149-58.
4. Mostafa MH, Helmi S, Badawi A, Tricker AR, Spiegelhalder B, Preussmann R. Nitrate, nitrite and volatile nitroso compounds in the urine of Schistosoma mansoni infected patients. Carcinogenesis 1994;15:619–25.
5. Krishna G, Kropko ML, Theiss JC. Dimethylnitrosamine-induced micronucleus formation in mouse bone marrow and spleen. Mutat Res 1990;242:345–51.
6. Zhang W, Yin L, Tao X, Xu L, Zheng L, Han X, et al. Dioscin alleviates dimethylnitrosamine-induced acute liver injury through regulating apoptosis, oxidative stress and inflammation. Environ Toxicol Pharmacol 2016;45:193–201.
7. Hussain MSB, Hiremath MB. Evulation of in vitro antioxidant, anti-inflammatory activities of aegle marmelos leaf extracts. Asian J Pharm Clin Res 2020;13:209-13.
8. Al-Attar AM, Al-Rethea HA. Chemoprotective effect of omega-3 fatty acids on thioacetamide-induced hepatic fibrosis in male rats. Saudi J Biol Sci 2017;24:956–65.
9. Reddy JK, Rao MS. Lipid metabolism and liver inflammation. II. Fatty liver disease and fatty acid oxidation. Am J Physiol Liver Physiol 2006;290:852–8.
10. Barth SP. Microanatomy and function of the spleen. Adv Anat Embryol Cell Biol 2000;151:1–101.
11. Little MH. Regrow or repair: potential regenerative therapies for the kidney. J Am Soc Nephrol 2006;17:2390–401.
12. Weibrich G, Kleis WK, Hafner G, Hitzler WE. Growth factor levels in plateletrich plasma and correlations with donor age, sex, and platelet count. J Craniomaxillofac Surg 2002;30:97–102.
13. Bielecki TM, Gazdzik TS, Arendt J, Szczepanski T, Krol W, Wielkoszynski T. Antibacterial effect of autologous platelet gel enriched with growth factors and other active substances: an in vitro study. J Bone Joint Surg Br 2007;89:417–20.
14. Cieslik Bielecka A, Bielecki T, Gazdzik TS, Arendt J, Krol W, Szczepanski T. Autologous platelets and leukocytes can improve healing of infected high-energy soft tissue injury. Transfus Apher Sci 2009;41:9–12.
15. Lyras DN, Kazakos K, Verettas D, Polychronidis A, Tryfonidis M, Botaitis S, et al. The influence of platelet-rich plasma on angiogenesis during the early phase of tendon healing. Foot Ankle Int 2009;30:1101–6.
16. Kark LR, Karp JM, Davies JE. Platelet releasate increases the proliferation and migration of bone marrow-derived cells cultured under osteogenic conditions. Clin Oral Implants Res 2006;17:321–7.
17. Abd El dayem SM, Zaazaa AM, Foda FM, Abd El Aty HE. Quercetin mitigates toxicity and oxidative stress motivated by bisphenol A in the liver of male rats. Int J Pharm Pharm Sci 2016;8:306-10.
18. Watzl B. Anti-inflammatory effects of plant-based foods and of their constituents. Int J Vitam Nutr Res 2008;78:293–8.
19. Zhang M, Swarts SG, Yin L, Liu C, Tian Y, Cao Y, et al. Antioxidant properties of quercetin. Adv Exp Med Biol 2011;701:283–9.
20. Bureau G, Longpre F, Martinoli MG. Resveratrol and quercetin, two natural polyphenols, reduce apoptotic neuronal cell death induced by neuroinflammation. J Neurosci Res 2008;86:403–10.
21. Tlaskalova Hogenova H, Stepankova R, Kozakova H, Hudcovic T, Vannucci L, Tuckova L, et al. The role of gut microbiota (commensal bacteria) and the mucosal barrier in the pathogenesis of inflammatory and autoimmune diseases and cancer: contribution of germ-free and gnotobiotic animal models of human diseases. Cell Mol Immunol 2011;8:110–20.
22. Yin YN, Yu QN, Liu XW, Lu FG. Effects of four Bifidobacteria on obesity in high-fat diet-induced rats. World J Gastroenterol 2010;16:3394–401.
23. Ley RE, Bäckhed F, Turnbaugh P, Lozupone CA, Knight RD, Gordon JI. Obesity alters gut microbial ecology. Proc Natl Acad Sci USA 2005;102:11070–5.
24. Mouria M, Gukovskaya AS, Jung Y, Buechler P, Hines OJ, Reber HA, et al. Food?derived polyphenols inhibit pancreatic cancer growth through mitochondrial cytochrome C release and apoptosis. Int J Cancer 2002;98:761–9.
25. Paolillo R, Carratelli CR, Rizzo A. Effect of resveratrol and quercetin in experimental infection by salmonella enterica serovar typhimurium. Int Immunopharmacol 2011;11:149–56.
26. Rao GS. N-nitrosamines from drugs and nitrite: a potential source of chemical carcinogens in humans? Pharmacol Intern 1980;1:187–90.
27. Wang J, Pan Y, Hong Y, Zhang Q, Wang X, Kong L. Quercetin protects against cadmium-induced renal uric acid transport system alteration and lipid metabolism disorder in rats. J Evidence Based Complementary Altern Med 2012;1-14.
28. Vendruscolo CP, Carvalho AM, Moraes LF, Maia L, Queiroz DL, Watanabe MJ, et al. Evaluating the effectiveness of different protocols for preparation of platelet rich plasma for use in equine medicine. Pesquisa Veterinaria Brasileira 2012;32:106-10.
29. Alves SBCR, Monteiro MPG, Pires NR, Santos AVP. Metodologia para obtenção do plasma rico em plaquetas: Estudo preliminar. Rev Eletron Novo Enfoque 2012;15:83-9.
30. Junior RR, Negriros RM, Elias FM, Jorge WA. The use of bone graft with plate rich plasma in the healing of bone defects. Rev Odontol Univ Cidade Sao Paulo 2008;20:295-300.
31. Usnayo MJG, Andrande LEC, Alarcon RT, Oliveira JC, Silva GMF, Bendet I, et al. Study of the frequency of HLA-DRB1 alleles in Brazilian patients with rheumatoid arthritis. Rev Bras Reumatol 2011;51:474-83.
32. Bakacak M, Bostanci MS, ?nanc F, Yaylali A, Serin S, Attar R, et al. Protective effect of platelet-rich plasma on experimental ischemia/reperfusion injury in rat ovary. Gynecol Obstet Investig 2016;81:225–31.
33. Franciele D, Malin H, Parmis B, Love T, Vinicius FB, Jefferson B, et al. Effect of platelet-rich plasma on rat achilles tendon healing is related to microbiota. Acta Orthop 2017;88:416-21.
34. Boussarie D. Hematologie des rongeurs et lagomorphes de compagnie. Bulletin de l'Academie Veterinaire de France 1999;72:209–16.
35. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265-75.
36. Satoh K. Serum lipid peroxide in cerebrovascular disorders determined by a new colorimetric method. Clin Chim Acta 1978;90:37-43.
37. Aebi H. Catalase in vitro. Methods Enzymol 1984;105:121–6.
38. Nishikimi M, Roa NA, Yogi K. The occurrence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen. Biochem Bioph Res Common 1972;46:849–54.
39. Fossati P, Prencipe L, Berti G. Use of 3,5-dichloro-2-hydroxybenzenesulfonic acid/4-aminophenazone chromogenic system in direct enzymic assay of uric acid in serum and urine. Clin Chem 1980;26:227–31.
40. Whicher J. C-eactive protein. In: Clinical laboratory diagnostics: use and assessment of clinical laboratory results/ed. by Lother Thomas. 1st Edition. Frankfurt/Main; 1998. p. 700-10.
41. Massaia M, Borrione P, Attisano C, Barral P, Beggiato E, Montacchini L, et al. Dysregulated FAS and Bcl-2 expression leading to enhanced apoptosis in T cells of multiple myeloma patients. Blood 1995;85:3679–87.
42. Bancroft JD, Gamble M. Theory and practice of histological techniques. 6th ed. Churchill Livingstone-Elsevier; 2008. p. 433-69.
43. Bancroft JD, Stevens A, Turner DR. Theory and practice of histological techniques. 4th ed. Churchill Livingstone Philadelphia PA USA; 1996. p. 25-90.
44. Cesta MF. Normal structure, function, and histology of the spleen. Toxicol Pathol 2006;34:455–65.
45. Melo ES, Barbeiro HV, Ariga S, Goloubkova T, Curi R, Valasco IT, et al. Immune cells and oxidative stress in the endotoxin tolerance mouse model. Braz J Med Biol Res 2010;43:57–67.
46. Liu Q, Raina AK, Smith MA, Sayre LM, Perry G. Hydroxynonenal, toxic carbonyls, and Alzheimer's disease. Mol Aspects Med 2003;24:305–13.
47. Halliwell B, Gutteridge JMC. Oxygen toxicity, oxygen radicals, transition metals, and disease. Biochem J 1984;218:1–14.
48. El-Shenawy NS, AL-Harbi MS, Hamza RZ. Effect of vitamin E and selenium separately and in combination on biochemical, immunological and histologicalchanges induced by sodium azide in male mice. Experim Toxicol Pathol 2014;67:65–76.
49. Hamza RZ, Ismail HAA, Khaled HE, El-Shenawy NS. Antioxidant effect of Lcysteineon sodium-valproate-induced oxidative stress in rat liver: biochemical andul trastructural approaches. Toxicol Environ Chem 2015;97:1041–56.
50. Cheeseman KH, Slater TF. An introduction to free radical biochemistry. Br Med Bull 1993;49:481–93.
51. Hesami Z, Jamshidzadeh A, Ayatollahi M, Geramizadeh B, Farshad O, Vahdati A. Effect of platelet-rich plasma on CCl4-induced chronic liver injury in male rats. Int J Hepatol 2014;932930:1-7.
52. Martins RP, Hartmann DD, de Moraes JP, Soares FA, Puntel GO. Platelet-rich plasma reduces the oxidative damage determined by a skeletal muscle contusion in rats. Platelets 2016;27:784–90.
53. Tohidnezhad M, Wruck CJ, Slowik A, Kweider N, Beckmann R, Bayer A, et al. Role of platelet-released growth factors in detoxification of reactive oxygen species in osteoblasts. Bone 2014;65:9–17.
54. Kajstura J, Fiordaliso F, Andreoli AM, Li B, Chimenti S, Medow MS, et al. IGF1 overexpression inhibits the development of diabetic cardiomyopathy and angiotensin II-mediated oxidative stress. Diabetes 2001;50:1414–24.
55. Zhou YJ, Yang HW, Wang XG, Zhang H. Hepatocyte growth factor prevents advanced glycation end products induced injury and oxidative stress through a PI3K/Akt dependent pathway in human endothelial cells. Life Sci 2009;85:670–7.
56. Zheng L, Ishii Y, Tokunaga A, Hamashima T, Shen J, Zhao QL, et al. Neuroprotective effects of PDGF against oxidative stress and the signaling pathway involved. J Neurosci Res 2010;88:1273–84.
57. Kaindl U, Eyberg I, Rohr Udilova N, Heinzle C, Marian B. The dietary antioxidants resveratrol and quercetin protect cells from exogenous pro-oxidative damage. Food Chem Toxicol 2008;46:1320–6.
58. Kumar R, Kaur K, Uppal S, Mehta SK. Ultrasound processed nanoemulsion: a comparative approach between resveratrol and resveratrol cyclodextrin inclusion complex to study its binding interactions, antioxidant activity and UV light stability. Ultrason Sonochem 2017;37:478–89.
59. Lin HL, Parsels LA, Maybaum J, Hollenberg PF. N-nitrosodimethylamine-mediated cytotoxicity in a cell line expressing P4502E1: evidence for apoptotic cell death. Toxicol Appl Pharmacol 1999;157:117–24.
60. Varfolomeev E, Goncharov T, Vucic D. Roles of c-IAP proteins in TNF receptor family activation of NF-?B signaling. NF-kappa B. Methods Protoc 2015;1280:269–82.
61. Moussa M, Lajeunesse D, Hilal G, El Atat O, Haykal G, Serhal R, et al. Platelet-rich plasma (PRP) induces chondroprotection via increasing autophagy, anti-inflammatory markers, and decreasing apoptosis in human osteoarthritic cartilage. Exp Cell Res 2017;352:146–56.
62. Cho SY, Park SJ, Kwon MJ, Jeong TS, Bok SH, Choi WY, et al. Quercetin suppresses proinflammatory cytokines production through MAP kinases and NF-kappaB pathway in lipopolysaccharide-stimulated macrophage. Mol Cell Biochem 2003;243:153-60.
63. Martinez Florez S, Gutierrez Fernandez B, Sanchez Campos S, Gonzalez Gallego J, Tunon MJ. Quercetin attenuates nuclear factor-kappaB activation and nitric oxide production in interleukin-1beta-activated rat hepatocytes. J Nutr 2005;135:1359-65.
64. Indra MR, Karyono S, Ratnawati R, Malik SG. Quercetin suppresses inflammation by reducing ERK1/2 phosphorylation and NF kappa B activation in leptin-induced human umbilical vein endothelial cells (HUVECs). BMC Res Notes 2013;6:275.
65. Chang YC, Tsai MH, Sheu WH, Hsieh SC, Chiang AN. The therapeutic potential and mechanisms of action of quercetin in relation to lipopolysaccharide-induced sepsis in vitro and in vivo. PLoS One 2013;8:e80744.
66. Comalada M, Camuesco D, Sierra S, Ballester I, Xaus J, Galvez J, et al. In vivo quercitrin anti-inflammatory effect involves release of quercetin, which inhibits inflammation through the downregulation of NF-B pathway. Eur J Immunol 2005; 35:584-92.
67. Min YD, Choi CH, Bark H, Son HY, Park HH, Lee S, et al. Quercetin inhibits expression of inflammatory cytokines through attenuation of NF-?B and p38 MAPK in HMC-1 human mast cell line. Inflamm Res 2007;56:210-5.
68. Nair MP, Mahajan S, Reynolds JL, Aalinkeel R, Nair H, Schwartz SA, et al. The flavonoid quercetin inhibits proinflammatory cytokine (tumor necrosis factor-alpha) gene expression in normal peripheral blood mononuclear cells via modulation of the NF-kappa beta system. Clin Vac Immunol 2006;13:319-28.
69. Kang CH, Choi YH, Moon SK, Kim WJ, Kim GY. Quercetin inhibits lipopolysaccharide-induced nitric oxide production in BV2 microglial cells by suppressing the NF?B pathway and activating the Nrf2-dependent HO-1 pathway. Int Immunopharmacol 2013;17:808-13.
70. Patel N, Joseph C, Corcoran GB, Ray SD. Silymarin modulates doxorubicin-induced oxidative stress, Bcl-xL and p53 expression while preventing apoptotic and necrotic cell death in the liver. Toxicol Appl Pharmacol 2010;245:143-52.
71. Weng SY, Yang CY, Li CC, Sun TP, Tung SY, Yen JJY, et al. Synergism betweenp53 and Mcl-1 in protecting from hepatic injury, fibrosis and cancer. J Hepatol 2011; 54:685–94.
72. Lee TY, Chang HH, Wang GJ, Chiu JH, Yang YY, Lin HC. Water-soluble extract of Salbia miltiorrhiza ameliorates carbon tetrachloride-mediated hepatic apoptosis in rats. J Pharm Pharmacol 2006;58:659–65.
73. Kong M, Ba M, Liang H, Ma L, Yu Q, Yu T, et al. 5'-Aza-dC sensitizes paraquat toxic effects on PC12 cell. Neurosci Lett 2012;524:35–9.
74. Zhang C, Wang F, Zhang Y, Kang Y, Wang H, Si M, et al. Celecoxib prevents pressure overload-induced cardiac hypertrophy and dysfunction by inhibiting inflammation, apoptosis and oxidative stress. J Cell Mol Med 2016;20:116–27.
75. Salem NA, Hamza A, Alnahdi H, Ayaz N. Biochemical and molecular mechanisms of platelet-rich plasma in ameliorating liver fibrosis induced by dimethyl nitrosourea. Cell Physiol Biochem 2018;47:2331–9.
76. Kim DH, Je YJ, Kim CD, Lee YH, Seo YJ, Lee JH, et al. Can platelet-rich plasma be used for skin rejuvenation? Evaluation of the effects of platelet-rich plasma on human dermal fibroblast. Ann Dermatol 2011;23:424–31.
77. Wu Z, Yu Y, Niu L, Fei L, Pan S. IGF-1 protects tubular epithelial cells during injury via activation of ERK/MAPK signaling pathway. Sci Rep 2016;6:28066.
78. Iwayama H, Ueda N. Role of mitochondrial Bax, caspases, and MAPKs for ceramide-induced apoptosis in renal proximal tubular cells. Mol Cell Biochem 2013;379:37–42.
79. Zhang J, Yang J, Liu Y. Role of Bcl-xL induction in HGF-mediated renal epithelial cell survival after oxidant stress. Int J Clin Exp Pathol 2008;1:242–53.
80. Xiaorong C, Cheng L, Yunfei l, Zongguo Y, Zhen L, Qingnian X, et al. Paeoniflorin regulates macrophage activation in dimethylnitrosamine-induced liver fibrosis in rats. BMC Complement Altern Med 2012;12:254.
81. Sampson S, Gerhardt M, Mandelbaum B. Platelet-rich plasma injection grafts for musculoskeletal injuries: a review. Curr Rev Musculoskelet Med 2008;1:165.
82. Camuesco D, Comalada M, Rodriguez Cabezas ME, Nieto A, Lorente MD, Concha A, et al. The intestinal anti?inflammatory effect of quercitrin is associated with an inhibition in iNOS expression. Br J Pharmacol 2004;143:908–18.
83. Galvez J, De la Cruz JP, Zarzuelo A, Sanchez de Medina F, Jimenez J, Sanchez de la Cuesta F. Oral administration of quercitrin modifies intestinal oxidative status in rats. Gen Pharmacol 1994;25:1237–43.
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