|Year : 2018 | Volume
| Issue : 5 | Page : 1057-1064
|Plantago major protects against cisplatin-induced renal dysfunction and tissue damage in rats
Soghra Parhizgar1, Sara Hosseinian2, Mohammad Soukhtanloo3, Alireza Ebrahimzadeh Bideskan4, Mousa-Al-Reza Hadjzadeh2, Samira Shahraki1, Zahra Samadi Noshahr1, Nazanin Entezari Heravi1, Milad Haghshenas1, Abolfazl Khajavi Rad2
1 Department of Physiology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
2 Department of Physiology, School of Medicine; Neurogenic Inflammation Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
3 Department of Biochemistry, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
4 Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
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|Date of Submission||13-Sep-2017|
|Date of Acceptance||10-Oct-2017|
|Date of Web Publication||26-Oct-2018|
| Abstract|| |
The aim of the present study was to determine the effect of Plantago major (P. major) on cisplatin-induced kidney injury in the rat. Cisplatin was injected on the 6th day of the experiment. Animals were treated with P. major extract (300, 600, and 1200 mg/kg) and Vitamin E for five days before and two weeks after cisplatin administration. Cisplatin caused a significant decrease in glomerular filtration rate (GFR), urine osmolarity, and urinary excretion rate of potassium, but significant increase in the kidney index and histological damage compared with the control group. Administration of Vitamin E and P. major (300 and 600 mg/kg) significantly increased GFR compared to cisplatin group. Furthermore, urine osmolarity in Vitamin E and P. major (600 mg/kg) groups were significantly elevated compared to the cisplatin group. P. major (600 mg/kg) significantly increased the urinary excretion rate of potassium compared with cisplatin group. Furthermore, all doses of P. major and Vitamin E significantly attenuated the percentage of kidney tissue damage compared to the cisplatin group. However, only P. major (600 mg/kg) and Vitamin E treated rats showed a significant reduction in the kidney index. This study revealed that P. major extract in a dose-dependent manner provides protection against renal damage induced by cisplatin.
|How to cite this article:|
Parhizgar S, Hosseinian S, Soukhtanloo M, Bideskan AE, Hadjzadeh MA, Shahraki S, Noshahr ZS, Heravi NE, Haghshenas M, Rad AK. Plantago major protects against cisplatin-induced renal dysfunction and tissue damage in rats. Saudi J Kidney Dis Transpl 2018;29:1057-64
|How to cite this URL:|
Parhizgar S, Hosseinian S, Soukhtanloo M, Bideskan AE, Hadjzadeh MA, Shahraki S, Noshahr ZS, Heravi NE, Haghshenas M, Rad AK. Plantago major protects against cisplatin-induced renal dysfunction and tissue damage in rats. Saudi J Kidney Dis Transpl [serial online] 2018 [cited 2019 Feb 21];29:1057-64. Available from: http://www.sjkdt.org/text.asp?2018/29/5/1057/243960
| Introduction|| |
Cisplatin is one of the most effective anticancer drugs and is widely used for treatment of a variety of cancers including the head and neck, testicular, breast, ovarian, bladder, and small-cell lung malignancies. Although, the clinical use of cisplatin is often limited due to its nephrotoxicity, which mainly develops in the proximal convoluted tubules. Nephrotoxicity is characterized by the signs of renal injury, including increased serum creatinine levels and uremia, elevation in urine output, increase in the products of lipid peroxidation, hypomagnesemia, hypokalemia, and reduced glomerular filtration rate (GFR).,, The mechanisms of cisplatin-induced nephrotoxicity are still not fully understood, however, several mechanisms have been considered including hypoxia, inflammation, oxidative stress, and cellular necrosis or apoptosis. Antioxidants are substances that have the ability to protect the body from damages caused by free radicalinduced oxidative stress. Some medicinal plants contained pharmacologically useful and antioxidant compounds and proved to be effective against various human ailments. Therefore, use of the medicinal herbs has increased dramatically in the past years. Plantago major (P. major) is a perennial plant belongs to the Plantaginaceae family. A wide ranges of biological activities have been found from the plant including analgesic, immunoenhancing, anti-inflammatory, antioxidant, anti-tumor, anti-hypertensive, anti-diabetic, anti-microbial, gastroprotective, and hepatoprotective properties. The aim of this study was to investigate the possible protective effects of P. major on kidney function and renal tissue damage in cisplatin-induced nephrotoxicity in the rat.
| Materials and Methods|| |
Cisplatin was purchased from the Mylan company (Greece). Vitamin E was purchased from 22 Bahman Pharmacy (Iran). P. major whole plant was obtained from the campus of Imam Reza hospital, Khorasan Razavi Province, Mashhad, Iran. All other reagents and solvents were of a high analytical grade.
P. major whole plants were collected from the campus of Imam Reza Hospital, Khorasan Razavi Province, Mashhad, Iran, and was recognized by botanists in the herbarium of Mashhad Ferdowsi University. For the preparation of the hydroalcoholic extract, P. major whole plant washed, dried, and powdered. Fifty grams of the powder was extracted in a Soxhlet extractor with ethanol (70% v/v). After the extraction, the solution was purified using a rotary vacuum evaporator and kept at 4°C until use.
Sixty adult male Albino Wistar rats weighing 250–300 g obtained from the Central Animal House of the School of Medicine, Mashhad University of Medical Sciences. The rats were housed at 22°C ± 2°C and standard condition of 12 h light/dark cycle with free access to food pellets and water. All experiments were performed under the authority of the Mashhad University of Medical Sciences and the norms of International Animal Ethics were followed.
In this study, animals were randomly divided into six groups of 10 each:
- Control group: Intraperitoneal (ip) injection of normal saline given on the 6th day of experiment.
- Cisplatin group: Injection of cisplatin (7 mg/kg, ip) given on the 6th day of experiment.
- Vitamin E + cisplatin group: Injection of Vitamin E (100 mg/kg, ip) for 20 consecutive days and injection of cisplatin on the 6th day of experiment.
- Extract-300 + cisplatin group: Administration of P. major extract (300 mg/kg) in drinking water for 20 consecutive days and injection of cisplatin on the 6th day of experiment.
- Extract-600 + cisplatin group: Administration of P. major extract (600 mg/kg) in drinking water for 20 consecutive days and injection of cisplatin on the 6th day of experiment.
- Extract-1200 + cisplatin group: Administration of P. major extract (1200 mg/kg) in drinking water for 20 consecutive days and injection of cisplatin on the 6th day of experiment.
Blood and 24-h urine samples were collected on days 0, 12, and 21 of the study. Two weeks after cisplatin injection, all animals were humanely killed and both kidneys were quickly removed. The right kidneys were fixed in 10% buffered formalin for histopathological examination. Blood samples were centrifuged at 4000 g for 10 min, and serums were stored at -20°C to be assayed later.
Kidney function tests
Serum and urine levels of creatinine were measured colorimetrically using a photometer (Convergys® 100 Biochemistry Analyser, Germany) and commercial diagnostic kits (Pars Azmoon Company, Tehran, Iran). Urineosmolarity was measured by a cryoscopic osmometer (Osmomat® 030, Germany). Urine potassium (K+) concentration was determined by using an electrolyte analyzer (AC 9800, China). Creatinine clearance (GFR) was calculated based on 24-h urine volume, serum, and urine creatinine concentration by:
Urinary excretion rate of K+ (Exck+) was calculated based on 24-h urine volume and urine K+ level by:
Urinary excretion rate of potassium = Urine concentration × Urine output
The rat’s kidney tissues from all groups were fixed in 10% formaldehyde, dehydrated in graded alcohol and embedded in paraffin. 5 μ sections were obtained, and slides were stained with hematoxylin and eosin for light microscopic analyses. The slides were evaluated by a pathologist who was blinded to the treatment groups. Renal tissue sections were graded as follows: (0) no damage; (1) patchy isolated unicellular necrosis; (2) tubular necrosis <25%; (3) tubular necrosis between 25% and 50% and (4) tubular necrosis >50%.
Kidney index was calculated based on the body weight of rats on day 21 and kidney weight by:
| Statistical Analysis|| |
All analyses were performed using Statistical Package for the Social Sciences program (SPSS) version 20.0 (SPSS Inc., Chicago, Illinois, USA). The data were expressed as Mean ± standard error of the mean. For each data group, the differences between days 12 and 0 were calculated. Differences between groups means were estimated using a one-way analysis of variance followed by least significant difference test for multiple comparisons. A P <0.05 was considered as statistically significant.
| Results|| |
Effect of P. major extract and Vitamin E on kidney function parameters
In the current study, kidney function tests were examined on the 1st, 12th, and 21st days of experiment. However, because there was no significant alteration in these parameters between cisplatin group and other treated animals on the 21th day, the data were not shown. As shown in [Table 1], cisplatin caused a significant decrease in the GFR as compared to control group (P <0.001). However, treatment of rats with Vitamin E and P. major extract (300 and 600 mg/kg) resulted in a significant increase in the GFR compared to the cisplatin group (P <0.01) [Table 1].
|Table 1: The effect of P. major extract and vitamin E on GFR, urinary excretion rate of potassium and urine osmolarity in all of experimental groups.|
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Urinary excretion rate of potassium in cisplatin group was significantly lower than control group (P <0.001). However, administration of P. major extract at the dose of 600 mg/kg significantly increased the urinary excretion rate of potassium compared to cisplatin-treated animals (P < 0.05) [Table 1].
Injection of cisplatin resulted in a significant decrease of urine osmolarity in cisplatin group compared to control group (P <0.001). However, Vitamin E and P. major extract at the dose of 600 mg/kg could significantly enhance the urine osmolarity compared to cisplatin group (P <0.05) [Table 1].
Effect of P. major extract and Vitamin E on the kidney injury
Renal tissue sections of the control group showed normal morphology [Figure 1]. A single dose administration of cisplatin resulted in a significant tubular and interstitial damage compared with those of control group (P <0.001) [Figure 1] and [Figure 2]. In epithelial cells of cisplatin-treated rats, desquamation, vacuolation and necrosis were observed. Hyaline casts and necrotic epithelial cells in luminal space of kidney tubules and intertubular hemorrhage were also observed in cisplatin group [Figure 1]. Treatment with P. major extract (300, 600, and 1200 mg/kg) and Vitamin E significantly improved the percent of renal histopathological changes compared with the cisplatin group (P <0.001) [Figure 1].
|Figure 1: Light microscopy of renal sections from different group of animals. Control group, showing normal morphology. Cisplatin group, showing necrosis in tubules, hyaline casts in tubules, desquamation, congestion and swelling in tubules and intertubular hemorrhage, vitamin E and P. major treated rats showing mild tubular and interstitial changes (hematoxylin and eosin, ×200).|
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|Figure 2: Percentage of renal tissue damage in all experimental groups of animal. Values are the Mean±SEM. The data were analyzed using one-way ANOVA and post hoc LSD. A significant difference was considered as P <0.05.|
***P <0.001 compared to control group on day 21. ###P <0.001 compared to cisplatin group on day 21.
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Kidney index of cisplatin-treated rats was significantly higher than those of the control animals (P <0.001). The administration of vitamin E and P. major extract (600 mg/kg) could significantly decreased the kidney index compared with the cisplatin group (P <0.01 and P<0.05, respectively) [Figure 3].
|Figure 3. Kidney index in all experimental groups of animal. Values are the Mean±SEM. The data were analyzed using one-way ANOVA and post hoc LSD. A significant difference was considered as P <0.05.|
***P<0.001 compared to control group on day 21, #P <0.05 and ##P <0.01 compared to cisplatin group on day 21.
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| Discussion|| |
The present results revealed that P. major extract in a dose-dependent manner showed considerable protective effects on the kidney dysfunction and tissue damage induced by cisplatin. Cisplatin is a potent nephrotoxic anticancer drug but the exact mechanism of its nephrotoxicity in not clearly known. Different studies have demonstrated that oxidative stress, inflammation, and apoptosis are closely associated with cisplatin-induced kidney injury.
In recent decades, herbal remedies have attracted great attention, and plant-derived compounds are increasingly used for the treatment of kidney disorders.,, P. major is a plant with a wide range of biological activities and various pharmacologic properties. The present work was planned to investigate the possible mechanisms of P. major extract against cisplatin-induced renal toxicity in the rat. We demonstrated that administration of a single dose of cisplatin caused significant change in the GFR that is in agreement with Pabla and Dong and Sanchez-Gonzalez et al investigations and confirmed the accuracy of our experiment in induction of kidney dysfunction., However, the current results showed that the administration of P. major extract at 300 and 600 mg/kg significantly increased the GFR. Various evidences indicate that cisplatin through the generation of reactive oxygen species and production of vasoconstrictor substances causes damages on glomerular compartments and contributes to decline in GFR., In this regard, the ameliorating effect of P. major extract on GFR might be due to its antioxidant and free radical scavenging properties. In addition, based on our results, cisplatin in accordance with Knight et al and Nematbakhsh et al findings caused a significant change in urine osmolarity and urinary excretion rate of potassium compared with the control group., However, in our study P. major extract at 600 mg/kg was significantly able to increase the urine osmolarity and urinary excretion rate of potassium compared with the cisplatin group. The possible mechanisms by which cisplatin affects the tubular transport of potassium are direct inhibition of Na+-K+ ATPase pump in the basolateral membrane of proximal tubular epithelial cells and inhibition of mitochondrial synthesis of ATP as well., Furthermore, the increase in urine output and dysfunction of sodium and potassium transporters might be involved., Possibly, P. major extract via its the antioxidant action may improve the mitochondrial function and increase ATP synthesis. In the present study, cisplatin caused severe glomerular and tubular damage and significantly increased the kidney index compared with the control animals. These structural changes have also been reported by other investigators.,, According to the present results, administration of P. major extract at 300, 600, and 1200 mg/kg significantly decreased the cisplatin-induced renal morphological changes. Meanwhile, the P. major extract could decrease the renal edema only at the dose of 600 mg/kg. Hussan et al described that P. major extract ameliorated the liver injury in acetaminophen treated rats. Numerous studies indicate that cisplatin via the production of oxygen free radicals and induction of inflammatory responses and apoptosis or necrosis causes multiple damages in renal tissue. On the same basis, the beneficial effects of the P. major extract against cisplatin-induced renal tissue damage are possibly related to its cyto-protective, antioxidant, and anti-inflammatory effects in the kidney.
| Conclusion|| |
The present investigation showed the ameliorating action of P. major extract on renal dysfunction and tissue damage-induced by cisplatin in a dose-dependent manner. However, it needs more investigations to clarify mechanisms involved in P. major action on cisplatin-induced kidney toxicity.
| Acknowledgment|| |
This study was part of a Master of Science thesis and was financially supported by the Research Council of Mashhad University of Medical Sciences, Mashhad, Iran.
We consider Dr. Sara Hosseinian as a first coauthor of this article because of the tremendous work and support given for this study.
Conflict of interest: None declared.
| References|| |
Katzung B, Masters SB, Trevor AJ: Basic and Clinical Pharmacology; NewYork: McGraw-Hill. 2012:1108.
Kröning R, Lichtenstein AK, Nagami GT. Sulfur-containing amino acids decrease cisplatin cytotoxicity and uptake in renal tubule epithelial cell lines. Cancer Chemother Pharmacol 2000;45:43-9.
Hartmann JT, Kollmannsberger C, Kanz L, Bokemeyer C. Platinum organ toxicity and possible prevention in patients with testicular cancer. Int J Cancer 1999;83:866-9.
Hosseinian S, Khajavi Rad A, Hadjzadeh MA, et al. The protective effect of nigella sativa against cisplatin-induced nephrotoxicity in rats. Avicenna J Phytomed 2016;6:44-54.
Kintzel PE. Anticancer drug-induced kidney disorders. Drug Saf 2001;24:19-38.
Ramesh G, Reeves WB. TNF-alpha mediates chemokine and cytokine expression and renal injury in cisplatin nephrotoxicity. J Clin Invest 2002;110:835-42.
Kawai Y, Nakao T, Kunimura N, Kohda Y, Gemba M. Relationship of intracellular calcium and oxygen radicals to cisplatin-related renal cell injury. J Pharmacol Sci 2006;100:65-72.
Chirino YI, Pedraza-Chaverri J. Role of oxidative and nitrosative stress in cisplatin-induced nephrotoxicity. Exp Toxicol Pathol 2009;61:223-42.
Mahantesh SP, Gangawane AK, Patil CS. Free radicals, antioxidants, diseases and phytomedicines in human health: Future perspects. World Res J Med Aromat Plants 2012;1:6-10.
Samuelsen AB. The traditional uses, chemical constituents and biological activities of Plantago
major L. A review. J Ethnopharmacol 2000;71:1-21.
Núñez Guillén ME, da Silva Emim JA, Souccar C, Lapa AJ. Analgesic and anti-inflammatory activities of the aqueous extract of Plantago
major L. Pharm Biol 1997;35:99-104.
Gomez-Flores R, Calderon CL, Scheibel LW, et al. Immunoenhancing properties of Plantago
major leaf extract. Phytother Res 2000;14:617-22.
Hussan F, Mansor AS, Hassan SN, et al. Anti-inflammatory property of Plantago
major leaf extract reduces the inflammatory reaction in experimental acetaminophen-induced liver injury. Evid Based Complement Alternat Med 2015;2015:347861.
Parhizgar S, Hosseinian S, Hadjzadeh MA, et al. Renoprotective effect of Plantago
major against nephrotoxicity and oxidative stress induced by cisplatin. Iran J Kidney Dis 2016;10:182-8.
Ozaslan M, Karagoz ID, Kilic IH, et al. Effect of Plantago
major sap on Ehrlich ascites tumours in mice. Afr J Biotechnol 2009;8:955-59.
Kyi KK, Bwin M, Gwan S, Maung C, et al. Hypotensive property of Plantago
major Linn. Union Burma Life Sci J 1972;4:167-71.
Noor H, Juing M, Chee BJ, Kueh BL, Othman Z. Medicinal Properties of Plantago
major: Hypoglycaemic and male fertility studies. Pertanika J Trop Agric Sci 2000;23:29-35.
Sharifa AA, Neoh YL, Iswadi MI. Effects of methanol, ethanol and aqueous extract of Plantago
major on gram positive bacteria, gram negative bacteria and yeast. Ann Microsc 2008;8:42-4.
Phipps M, Mahmood A. Gastroprotective activity of P. Major in rats. Int J Trop Med 2006;1:36-9.
Türel I, Ozbek H, Erten R, et al. Hepato-protective and anti-inflammatory activities of Plantago
major L. Indian J Pharmacol 2009; 41:120-4.
Zhou H, Miyaji T, Kato A, et al. Attenuation of cisplatin-induced acute renal failure is associated with less apoptotic cell death. J Lab Clin Med 1999;134:649-58.
Yao X, Panichpisal K, Kurtzman N, Nugent K. Cisplatin nephrotoxicity: A review. Am J Med Sci 2007;334:115-24.
Mohebbati R, Shafei MN, Soukhtanloo M, et al. Adriamycin-induced oxidative stress is prevented by mixed hydro-alcoholic extract of nigella sativa and Curcuma longa
in rat kidney. Avicenna J Phytomed 2016;6:86-94.
Rad AK, Ghazi L, Boroushaki MT, et al. Effect of commercial (vimang) and hydro-alcoholic extract of Mangifera indica
(Mango) on gentamicin-induced nephrotoxicity in rat. Avicenna J Phytomed 2011;1:98-105.
Pabla N, Dong Z. Cisplatin nephrotoxicity: Mechanisms and renoprotective strategies. Kidney Int 2008;73:994-1007.
Sanchez-Gonzalez PD, Lopez-Hernandez FJ, Perez-Barriocanal F, Morales AI, Lopez-Novoa JM. Quercetin reduces cisplatin nephrotoxicity in rats without compromising its anti-tumour activity. Nephrol Dial Transplant 2011;26:3484-95.
Miller RP, Tadagavadi RK, Ramesh G, Reeves WB. Mechanisms of cisplatin nephrotoxicity. Toxins (Basel) 2010;2:2490-518.
Nath KA, Norby SM. Reactive oxygen species and acute renal failure. Am J Med 2000; 109:665-78.
Knight RJ, Collis MG, Yates MS, Bowmer CJ. Amelioration of cisplatin-induced acute renal failure with 8-cyclopentyl-1,3-dipropylxanthine. Br J Pharmacol 1991;104:1062-8.
Nematbakhsh M, Ashrafi F, Safari T, et al. Administration of Vitamin E and losartan as prophylaxes in cisplatin-induced nephrotoxi-city model in rats. J Nephrol 2012;25:410-7.
Halabe A, Wong NL, Sutton RA. Effect of chronic cisplatin administration on phosphate and glucose transport by the renal brush border membrane. Nephron 1991;57:197-200.
Lajer H, Kristensen M, Hansen HH, et al. Magnesium depletion enhances cisplatin-induced nephrotoxicity. Cancer Chemother Pharmacol 2005;56:535-42.
Francescato HD, Coimbra TM, Costa RS, Bianchi Mde L. Protective effect of quercetin on the evolution of cisplatin-induced acute tubular necrosis. Kidney Blood Press Res 2004;27:148-58.
Kishore BK, Krane CM, Di Iulio D, Menon AG, Cacini W. Expression of renal aquaporins 1, 2, and 3 in a rat model of cisplatin-induced polyuria. Kidney Int 2000;58:701-11.
Shafiee S, Ebrahimzadeh A, Rajaei Z, et al. Effects of aqueous-ethanolic extract of nigella sativa seeds (Black Cumin) and Vitamin E on cisplatin-induced nephrotoxicity in rat. Res J Med Plants 2016;10:295-302.
Ahangarpour A, Abdollahzade Fard A, Gharibnaseri MK, Jalali T, Rashidi I. Hydrogen sulfide ameliorates the kidney dysfunction and damage in cisplatin-induced nephrotoxicity in rat. Vet Res Forum 2014; 5:121-7.
Dr. Abolfazl Khajavi Rad
Department of Physiology, Faculty of Medicine, Mashhad University of Medical Sciences Mashhad
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