, Volume 26, Issue 3, pp 779–791 | Cite as

Ziziphus spina-christi leaf extract pretreatment inhibits liver and spleen injury in a mouse model of sepsis via anti-oxidant and anti-inflammatory effects

  • Mohamed A. Dkhil
  • Saleh Al-Quraishy
  • Ahmed E. Abdel Moneim
Original Article


Sepsis is a systemic response to infection that can result in acute hepatic and splenic damage. Ziziphus spina-christi (L.) is a wild tree used as a medicinal plant by ancient Egyptians. However, little is known about the mechanism underlying its effects on sepsis. The current study investigated the protective effects of a Z. spina-christi leaf extract (ZSCLE) on liver and spleen damage in a male C57BL/6 mouse model of sepsis, induced by cecal ligation and puncture (CLP). Prior to CLP, ZSCLE was administered daily for five consecutive days via oral gavage at doses of 100, 200, or 300 mg/kg. The mice were euthanized 9 h after CLP, and oxidative stress markers were measured (myeloperoxidase, lipid peroxidation, nitric oxide, and reduced glutathione). In addition, we investigated histological changes, anti-oxidant enzyme activities (superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase), cytokine levels, protein expression of nuclear factor-κB and inducible nitric oxide synthase (iNOS), and mRNA levels of mitogen-activated protein kinase (8, 9, and 14), iNOS, tumor necrosis factor-α, and interleukin-1β. Our results indicated that ZSCLE significantly and dose-dependently inhibited sepsis-induced liver and spleen injury. These results suggest that ZSCLE could provide a therapeutic agent for sepsis by inducing anti-inflammatory and anti-oxidant effects.


Ziziphus spina-christi Sepsis Mitogen-activated protein kinase Nuclear factor-κB Anti-oxidant 



Cecal ligation and puncture


Glyceraldehyde 3-phosphate dehydrogenase


Glutathione peroxidase


Glutathione reductase






Inducible nitric oxide synthase






Mitogen-activated protein kinase


Multi-organ dysfunction


Reduced nicotinamide adenine dinucleotide phosphate


Nuclear factor-κB


Nitric oxide


Reactive oxygen species


Tumor necrosis factor-α


Ziziphus spina-christi (L.)


Ziziphus spina-christi leaf extract



The authors would like to express their appreciation to the Deanship of Scientific Research at King Saud University for funding this study through the research group project No. RG-198.

Compliance with ethical standards

Conflict of interest

No potential conflict of interest was reported by the authors.

Supplementary material

10787_2017_439_MOESM1_ESM.jpg (190 kb)
Supplementary Figure: Effects of Ziziphus spina-christi leaf extract (ZSCLE) administration on the scoring index of histopathological evaluation in mice with cecal ligation and puncture (CLP)-induced sepsis. Values represent the mean ± SEM (n = 7). aSignificantly different from sham (p < 0.05); bSignificantly different from CLP (p < 0.05). (JPEG 189 kb)


  1. Abdel Moneim AE (2013) The neuroprotective effects of purslane (Portulaca oleracea) on rotenone-induced biochemical changes and apoptosis in brain of rat. CNS Neurol Disord Drug Targets 12:830–841 (CNSNDDT-EPUB-53920) CrossRefPubMedGoogle Scholar
  2. Abdel Moneim AE (2016) Indigofera oblongifolia prevents lead acetate-induced hepatotoxicity, oxidative stress, fibrosis and apoptosis in rats. PLoS ONE 11:e0158965. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Abraham E (2003) Nuclear factor-kappaB and its role in sepsis-associated organ failure. J Infect Dis 187(Suppl 2):S364–S369 (JID21050) CrossRefPubMedGoogle Scholar
  4. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126CrossRefPubMedGoogle Scholar
  5. Alici O, Kavakli HS, Koca C, Altintas ND, Aydin M, Alici S (2015) Value of caffeic acid phenethyl ester pretreatment in experimental sepsis model in rats. Mediat Inflamm 2015:810948. CrossRefGoogle Scholar
  6. Al-Olayan E, El-Khadragy M, Metwally D, Abdel Moneim A (2014) Protective effects of pomegranate (Punica granatum) juice on testes against carbon tetrachloride intoxication in rats. BMC Complement Altern Med 14:164. CrossRefPubMedPubMedCentralGoogle Scholar
  7. Amin A, Mahmoud-Ghoneim D (2009) Zizyphus spina-christi protects against carbon tetrachloride-induced liver fibrosis in rats. Food Chem Toxicol 47:2111–2119. CrossRefPubMedGoogle Scholar
  8. Andrades M, Ritter C, de Oliveira MR, Streck EL, Fonseca Moreira JC, Dal-Pizzol F (2011) Antioxidant treatment reverses organ failure in rat model of sepsis: role of antioxidant enzymes imbalance, neutrophil infiltration, and oxidative stress. J Surg Res 167:e307–e313 (pii:S0022-4804(09)00425-9) CrossRefPubMedGoogle Scholar
  9. Asaduzzaman M, Wang Y, Thorlacius H (2008) Critical role of p38 mitogen-activated protein kinase signaling in septic lung injury. Crit Care Med 36:482–488. CrossRefPubMedGoogle Scholar
  10. Bacanli M, Aydin S, Taner G, Goktas HG, Sahin T, Basaran AA, Basaran N (2016) Does rosmarinic acid treatment have protective role against sepsis-induced oxidative damage in Wistar Albino rats? Hum Exp Toxicol 35:877–886. CrossRefPubMedGoogle Scholar
  11. Bradley PP, Priebat DA, Christensen RD, Rothstein G (1982) Measurement of cutaneous inflammation: estimation of neutrophil content with an enzyme marker. J Invest Dermatol 78:206–209 (S0022-202X(15)46301-8) CrossRefPubMedGoogle Scholar
  12. Brown GC, Borutaite V (2007) Nitric oxide and mitochondrial respiration in the heart. Cardiovasc Res 75:283–290 (pii:S0008-6363(07)00147-2) CrossRefPubMedGoogle Scholar
  13. Carlberg I, Mannervik B (1985) Glutathione reductase Methods Enzymol 113:484–490PubMedGoogle Scholar
  14. Escames G, Lopez LC, Ortiz F, Lopez A, Garcia JA, Ros E, Acuna-Castroviejo D (2007) Attenuation of cardiac mitochondrial dysfunction by melatonin in septic mice. FEBS J 274:2135–2147 (EJB5755) CrossRefPubMedGoogle Scholar
  15. Grisham MB, Johnson GG, Lancaster JR Jr (1996) Quantitation of nitrate and nitrite in extracellular fluids. Methods Enzymol 268:237–246CrossRefPubMedGoogle Scholar
  16. Guizani N, Waly MI, Singh V, Rahman MS (2013) Nabag (Zizyphus spina-christi) extract prevents aberrant crypt foci development in colons of azoxymethane-treated rats by abrogating oxidative stress and inducing apoptosis. Asian Pac J Cancer Prev 14:5031–5035. CrossRefPubMedGoogle Scholar
  17. Jang JY, Lee SH, Shim H, Lee JG (2017) Serum oxygen radical activity and total antioxidation capacity are related with severities of surgical patient with sepsis: prospective pilot study. J Crit Care 39:131–136 (pii:S0883-9441(16)30731-6) CrossRefPubMedGoogle Scholar
  18. Kadioglu O et al (2016) Evaluating ancient Egyptian prescriptions today: anti-inflammatory activity of Ziziphus spina-christi. Phytomedicine 23:293–306 (S0944-7113(16)00022-2) CrossRefPubMedGoogle Scholar
  19. Kim SJ, Baek KS, Park HJ, Jung YH, Lee SM (2016) Compound 9a, a novel synthetic histone deacetylase inhibitor, protects against septic injury in mice by suppressing MAPK signalling. Br J Pharmacol 173:1045–1057. CrossRefPubMedPubMedCentralGoogle Scholar
  20. Ko SY et al (2015) Spinosin, a C-glucosylflavone, from Zizyphus jujuba var. spinosa Ameliorates Abeta1-42 oligomer-induced memory impairment in mice. Biomol Ther (Seoul) 23:156–164. CrossRefGoogle Scholar
  21. Kothari N et al (2011) Increased myeloperoxidase enzyme activity in plasma is an indicator of inflammation and onset of sepsis. J Crit Care 26(435):e431–e437 (pii:S0883-9441(10)00262-5) Google Scholar
  22. Liang N, Kitts DD (2015) Role of chlorogenic acids in controlling oxidative and inflammatory stress conditions. Nutrients. CrossRefPubMedPubMedCentralGoogle Scholar
  23. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  24. Michel CG, Nesseem DI, Ismail MF (2011) Anti-diabetic activity and stability study of the formulated leaf extract of Zizyphus spina-christi (L.) Willd with the influence of seasonal variation. J Ethnopharmacol 133:53–62 (pii:S0378-8741(10)00639-2) CrossRefPubMedGoogle Scholar
  25. Morgan MJ, Liu ZG (2011) Crosstalk of reactive oxygen species and NF-kappaB signaling. Cell Res 21:103–115. CrossRefPubMedGoogle Scholar
  26. Mubaraki MA, Hafiz TA, Al-Quraishy S, Dkhil MA (2017) Oxidative stress and genes regulation of cerebral malaria upon Zizyphus spina-christi treatment in a murine model. Microb Pathog 107:69–74. CrossRefPubMedGoogle Scholar
  27. Nishikimi M, Appaji N, Yagi K (1972) The occurrence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen. Biochem Biophys Res Commun 46:849–854CrossRefPubMedGoogle Scholar
  28. Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358CrossRefPubMedGoogle Scholar
  29. Osterbur K, Mann FA, Kuroki K, DeClue A (2014) Multiple organ dysfunction syndrome in humans and animals. J Vet Intern Med 28:1141–1151. CrossRefPubMedPubMedCentralGoogle Scholar
  30. Prauchner CA (2017) Oxidative stress in sepsis: pathophysiological implications justifying antioxidant co-therapy. Burns 43:471–485 (pii:S0305-4179(16)30400-4) CrossRefPubMedGoogle Scholar
  31. Rahim I et al (2017) Melatonin administration to wild-type mice and non-treated NLRP3 mutant mice share similar inhibition of the inflammatory response during sepsis. J Pineal Res. PubMedCrossRefGoogle Scholar
  32. Schulte W, Bernhagen J, Bucala R (2013) Cytokines in sepsis: potent immunoregulators and potential therapeutic targets—an updated view. Mediat Inflamm 2013:165974. CrossRefGoogle Scholar
  33. Shi C, Pamer EG (2011) Monocyte recruitment during infection and inflammation. Nat Rev Immunol 11:762–774. CrossRefPubMedPubMedCentralGoogle Scholar
  34. Thipparaboina R, Mittapalli S, Thatikonda S, Nangia A, Naidu VGM, Shastri NR (2016) Syringic acid: structural elucidation and co-crystallization. Cryst Growth Des 16:4679–4687. CrossRefGoogle Scholar
  35. Ursini F, Maiorino M, Gregolin C (1985) The selenoenzyme phospholipid hydroperoxide glutathione peroxidase. Biochim Biophys Acta 839:62–70CrossRefPubMedGoogle Scholar
  36. Wan SX, Shi B, Lou XL, Liu JQ, Ma GG, Liang DY, Ma S (2016) Ghrelin protects small intestinal epithelium against sepsis-induced injury by enhancing the autophagy of intestinal epithelial cells. Biomed Pharmacother 83:1315–1320 (pii:S0753-3322(16)30767-3) CrossRefPubMedGoogle Scholar
  37. Wang Y et al (2016) Berberine in combination with yohimbine attenuates sepsis-induced neutrophil tissue infiltration and multiorgan dysfunction partly via IL-10-mediated inhibition of CCR2 expression in neutrophils. Int Immunopharmacol 35:217–225 (pii:S1567-5769(16)30123-0) CrossRefPubMedGoogle Scholar
  38. Xiao X, Yang M, Sun D, Sun S (2012) Curcumin protects against sepsis-induced acute lung injury in rats. J Surg Res 176:e31–e39 (pii:S0022-4804(11)01992-5) CrossRefPubMedGoogle Scholar
  39. Yang M, Wu J, Martin CM, Kvietys PR, Rui T (2008) Important role of p38 MAP kinase/NF-kappaB signaling pathway in the sepsis-induced conversion of cardiac myocytes to a proinflammatory phenotype. Am J Physiol Heart Circ Physiol 294:H994–H1001. CrossRefPubMedGoogle Scholar
  40. Zhong W, Qian K, Xiong J, Ma K, Wang A, Zou Y (2016) Curcumin alleviates lipopolysaccharide induced sepsis and liver failure by suppression of oxidative stress-related inflammation via PI3K/AKT and NF-kappaB related signaling. Biomed Pharmacother 83:302–313 (pii:S0753-3322(16)30602-3) CrossRefPubMedGoogle Scholar
  41. Zhou J et al (2012) Hydrogen-rich saline reverses oxidative stress, cognitive impairment, and mortality in rats submitted to sepsis by cecal ligation and puncture. J Surg Res 178:390–400 (pii:S0022-4804(12)00062-5) CrossRefPubMedGoogle Scholar
  42. Zolali E, Hamishehkar H, Maleki-Dizaji N, Majidi Zolbanin N, Ghavimi H, Kouhsoltani M, Asgharian P (2014) Selenium effect on oxidative stress factors in septic rats. Adv Pharm Bull 4:289–293. PubMedPubMedCentralCrossRefGoogle Scholar
  43. Zolali E, Asgharian P, Hamishehkar H, Kouhsoltani M, Khodaii H (2015) Effects of gamma oryzanol on factors of oxidative stress and sepsis-induced lung injury in experimental animal model. Iran J Basic Med Sci 18:1257–1263. PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Zoology, College of ScienceKing Saud UniversityRiyadhSaudi Arabia
  2. 2.Department of Zoology and Entomology, Faculty of ScienceHelwan UniversityCairoEgypt

Personalised recommendations