HEPATOPROTECTIVE EFFECTS OF L-CITRULLINE AGAINST DOXORUBICIN-INDUCED LIVER DAMAGE IN RATS: AN ANALYSIS OF SERUM BIOMARKERS
Objective: The antineoplastic agent doxorubicin (DOX) is known for causing liver toxicity. Its metabolism in hepatocytes causes oxidative stress, which,
in turn, induces DNA damage, lipid peroxidation, ATP depletion, and apoptosis. L-citrulline (CIT), a commonly found agent in fruits like watermelon,
has piqued interest due to its antioxidant properties. In the body, CIT is converted to nitric oxide, which has been shown to mitigate hepatic injury
by scavenging free radicals, improving hepatic sinusoidal microcirculation, and inhibiting neutrophilic infiltration. This study aims to investigate CIT
ability to prevent DOX-induced hepatotoxicity.
Methods: A total of 20 Wistar rats were randomized to receive either DOX (10 mg/kg BW) or NaCl 0.9%. DOX-intoxicated group was further randomized
to either received low-dose CIT (300 mg/kg BW), high-dose CIT (600 mg/kg BW), or aquadest. CIT was given orally for 6 days and DOX through
intraperitoneal injection on days 4 and 5. Serum was obtained and hepatotoxicity was assessed with serum levels of aspartate aminotransferase
(AST), alanine aminotransferase (ALT), and gamma-glutamyl transferase (GGT). Statistical analysis was done with one-way ANOVA and Tukey’s test.
Results: Serum ALT, AST, and GGT were increased significantly compared to that of normal group. CIT administration in both the doses could decrease
the serum levels of ALT and AST significantly compared to that of DOX group. In this study, CIT in both the doses could reduce the serum levels of GGT
compared to that of DOX group though not statistically significant.
Conclusions: This study suggests that CIT exerts hepatoprotective effect, as evident by the attenuation of serum biomarkers.
anticancer molecular action, toxicity and novel drug delivery systems.
J Pharm Pharmacol 2013;65:157-70.
2. Thorn CF, Oshiro C, Marsh S, Hernandez-Boussard T, McLeod H,
Klein TE, et al. Doxorubicin pathways: Pharmacodynamics and
adverse effects. Pharmacogenet Genomics 2011;21:440-6.
3. Damodar G, Smitha T, Gopinath S, Vijayakumar S, Rao YA. An
evaluation of hepatotoxicity in breast cancer patients receiving injection
doxorubicin. Ann Med Health Sci Res 2014;4:74-9.
4. Yang XL, Fan CH, Zhu HS. Photo-induced cytotoxicity of malonic acid
[C(60)]fullerene derivatives and its mechanism. Toxicol In Vitro 2002;
5. Bhandari NR, Shewale AR, Kathe NJ, Shah AB, Painter JT. Association
between the use of doxorubicin and risk of developing hepatotoxicity
among cancer patients. Value Health 2016;19:A134.
6. El-Sayyad HI, Ismail MF, Shalaby FM, Abou-El-Magd RF, Gaur RL,
Fernando A, et al. Histopathological effects of cisplatin, doxorubicin and
5-flurouracil (5-FU) on the liver of male albino rats. Int J Biol Sci 2009;
7. Bengaied D, Ribeiro A, Amri M, Scherman D, Arnaud P. Reduction of
hepatotoxicity induced by doxorubicin. J Integr Oncol 2017;6:193.
8. Grigorian A, O’Brien CB. Hepatotoxicity secondary to chemotherapy.
J Clin Transl Hepatol 2014;2:95-102.
9. Bahri S, Zerrouk N, Aussel C, Moinard C, Crenn P, Curis E,
et al. Citrulline: From metabolism to therapeutic use. Nutrition 2013;
10. Papadia C, Osowska S, Cynober L, Forbes A. Citrulline in health and
disease. Review on human studies. Clin Nutr 2018;37:1823-8.
11. Rajapakse NW, Mattson DL. Role of L-arginine in nitric oxide
production in health and hypertension. Clin Exp Pharmacol Physiol
12. Taha MO, Simões MJ, Haddad MA, Capelato RC, Budny N,
Matsumoto AH, et al. L-arginine supplementation protects against
hepatic ischemia-reperfusion lesions in rabbits. Transplant Proc 2009;
13. Suzuki T, Morita M, Kobayashi Y, Kamimura A. Oral L-citrulline
supplementation enhances cycling time trial performance in healthy
trained men: Double-blind randomized placebo-controlled 2-way
crossover study. J Int Soc Sports Nutr 2016;13:1-8.
14. Morita M, Hayashi T, Ochiai M, Maeda M, Yamaguchi T, Ina K, et al.
Oral supplementation with a combination of L-citrulline and L-arginine
rapidly increases plasma L-arginine concentration and enhances NO
bioavailability. Biochem Biophys Res Commun 2014;454:53-7.
15. Moussaoui F, Binimbi A, Cottart CH, Amen N, Vamy M, Nivet-
Antoine V, et al. Hepatoprotective effect of citrulline against ischemiareperfusion
injury in rat. J Hepatol 2009;50:S68.
16. Jegatheesan P, Beutheu S, Ventura G, Nubret E, Sarfati G, Bergheim I,
et al. Citrulline and nonessential amino acids prevent fructose-induced
nonalcoholic fatty liver disease in rats. J Nutr 2015;145:2273-9.
17. Hayward R, Hydock DS. Doxorubicin cardiotoxicity in the rat: An
in vivo characterization. J Am Assoc Lab Anim Sci 2007;46:20-32.
18. Yi L, Lingshan G, Cui Y, Xiaoxing Y, Junnian Z. A preliminary study on
protective effect of L-citrulline against ischemia-reperfusion induced
gastric mucosal lesions in rat. Indian J Pharmacol 2012;44:31-5.
19. Dewanjee S, Joardar S, Bhattacharjee N, Dua TK, Das S, Kalita J,
et al. Edible leaf extract of ipomoea aquatica forssk. (Convolvulaceae)
attenuates doxorubicin-induced liver injury via inhibiting oxidative
impairment, MAPK activation and intrinsic pathway of apoptosis. Food
Chem Toxicol 2017;105:322-36.
20. Mohajeri M, Sahebkar A. Protective effects of curcumin against
doxorubicin-induced toxicity and resistance: A review. Crit Rev Oncol
21. Aulbach AD, Amuzie CJ. Biomarkers in Nonclinical Drug
Development. In: A Comprehensive Guide to Toxicology in Nonclinical
Drug Development. Elsevier; 2017. p. 447-71. Available from: http://
[Last accessed on 2018 Jun 26].
22. Boone L, Meyer D, Cusick P, Ennulat D, Bolliger AP, Everds N, et al.
Selection and interpretation of clinical pathology indicators of hepatic
injury in preclinical studies. Vet Clin Pathol 2005;34:182-8.
23. York MJ. Clinical Pathology. In: A Comprehensive Guide to Toxicology
in Nonclinical Drug Development. Elsevier; 2017 p. 325?74.
Available from: http://www.linkinghub.elsevier.com/retrieve/pii/
B9780128036204000141. [Last accessed on 2018 Jun 26].
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