Received 2018-06-16

Revised 2018-10-03

Accepted 2018-11-05

 Correspondence to: Farzane Zarifi,Legal Medicine Research Center, Tehran, Iran Telephone Number: +989173129507 Email Address : zarififarzane@rocketmail.com

GMJ.2018;7:e1258

www.gmj.ir

Introduction

Cisplatin, also named as cis-diamminedichloroplatinum (II), is an anti-neoplastic bifunctional (has properties of two different functional groups) alkylating agent, belonging to the group of platinum anti-tumor compounds. For example, cisplatin is commonly used in chemotherapy of various forms of cancer including head, neck, ovarian, bladder, and testicular cancers [1].

Despite cisplatin beneficial effects, it has considerable nephrotoxic and neurotoxic side effects that often limit its clinical use. For instance, renal proximal tubular toxicity is the major limitation of using cisplatin [2].

Neither prophylactic intensive hydration nor force diuresis has been effective in eliminating cisplatin toxicity [3].

DNA damage and apoptosis induced by cisplatin (oxidative stress) appear to be the mechanisms by which this drug produces renal injury. The oxidative stress resulting from increased free radical generation plays a key role in the pathogenesis of cisplatin-induced nephrotoxicity. Studies have shown that abnormal production of reactive oxygen species (ROS) can cause renal proximal tubule cellular damage and necrosis [4-10].

Corn-silk (CS), stigma/style of Zea mays Linne, is a famous traditional herbal medicine, which has been widely used for the treatment of edema, cystitis, gout, kidney stones, nephritis, and prostatitis [11, 12].

CS is known to have a wide range of pharmacological and biological activities [13].

This herb contains proteins, vitamins, carbohydrates, calcium, potassium, manganese and sodium salts, volatile oils, and steroids. The diuretic action of CS is partly due to its significant K content [14-16].

CS is also known to be rich in phenolic compounds such as anthocyanin, p-coumaric acid, vanillic acid, protocatechuic acid, derivatives of hesperidin and quercetin, and bounded hydroxycinnamic acid components composed of p-coumaric and ferulic acid. Phenolic compounds present in CS play an important role in its antioxidant and free radical scavenging capacity [12].

Methanol extracts of CS exerted antioxidant effects against lipid peroxidation [17].

Consumption of CS was also reported to be safe and has no adverse effects for humans [18].

In previous investigations, CS has been shown to have beneficial effects on nephrotoxicity via the amelioration of oxidative injury [19, 20].

This study was designed to investigate the possible protective effects of CS against cisplatin-induced nephrotoxicity in rat models.

Materials and Methods

Animals

Thirty-five rats aged about 10-12 weeks and with 220 ± 20g mass were obtained from our animal breeding center, Shiraz University of Medical Sciences, Shiraz, Iran.

Preparation of CS Extract

The CS was collected and authenticated by the department of pharmacology at Shiraz University of Medical Sciences. The plant was dried and powdered at room temperature. The methanol extraction was prepared by soaking the plant powder into 1000ml of 80% methanol.

Study Groups

The rats were randomly divided into five groups of seven each: C1 as the untreated control group, which received distilled water (1 ml/day) orally for eight consecutive days; group 2, the C2 received a single dose of cisplatin (5 mg/Kg, i.p.) on the first day of study. The CS group received 300 mg/kg/day of the CS extract orally. Groups 4 (CS1) and 5 (CS2) received 200 and 300 mg/kg/day of the CS extract orally, respectively, for eight consecutive days. CS1 and CS2 received a single dose of cisplatin (5mg/Kg, i.p.) post-CS extract dosage only on the first day.

Sample Collection and Biochemical Measurements

After nine days, all animals were anesthetized with ether; blood samples were prepared by cardiocentesis for measuring the serum levels of alanine transaminase (ALT) and aspartate transaminase (AST), blood urea nitrogen (BUN). The left kidney was removed, homogenized in cold potassium chloride solution (1.5%) to give a 10% homogenate, and used for measuring malondialdehyde (MDA). Lipid peroxidation was measured as the amount of MDA determined by the thiobarbituric acid reactive substance [21].

Histopathological Evaluations

All the kidneys were fixed by the 10% formaldehyde; then, we prepared 5 µm sections by microtome. All the slides were stained with hematoxylin and eosin. We examined the degree of the presence of congestion and degenerative cellular changes. The level of each pathological manifestation was graded according to the changes involved: none with 0, less than 20 % with 1, 21–40 % with 3, 61–80 % with 4, and greater than 80 % with 5. The sum of all numerical scores in each group was taken as the total histopathological score [22].

Statistical Analysis

Data are given as mean ± standard error of the mean. The statistically significant differences were assessed using one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparisons, and post hoc test. A P<0.05 was considered as significant level.

ResultEffects of CS on Cisplatin-Induced Lipid Peroxidation

As illustrated in Figure-1, the CS group animals showed no any significant difference compared to the C1 group, whereas the cisplatin-treated groups represented a considerable increase in their levels of MDA (P<0.01). Pre-treatment with CS exhibited a significant decrease in the MDA levels as compared to the values obtained after treatment with cisplatin alone (P<0.01).

Effects of CS on BUN Level

The levels of the BUN were examined in all groups. The cisplatin-treated groups showed a significant increase in the levels of BUN (P<0.01). The CS1 and CS2 animals pre-treated with CS followed by a single dose of cisplatin showed a significant decrease in the level of BUN compared to cisplatin alone (P<0.01, Figure-2).

Effects of CS on AST and ALT Levels

AST and ALT levels were higher in the cisplatin-treated groups when compared to the control group. In contrast, the CS2 rats revealed a significant decrease in the level of the same parameters in comparison to C2 (P<0.05). However, the hepatic function was improved in cisplatin-treated animals, which were pre-treated with CS (Figure-3).

Effect of CS on Histopathological Changes

Histological findings of the kidney from various treatment groups are presented in Figure-4. Treatment with cisplatin caused extensive tubular necrosis and desquamation. The pre-treatment with CS decreased the tubular necrosis compared to cisplatin alone.

Discussion

Cisplatin is considered as one of the active cytotoxic agents in the treatment of cancer. However, nephrotoxicity, hepatotoxicity, and neurotoxicity are considered as cisplatin’s side effects [23]. Previous researches have revealed that cisplatin induces nephrotoxicity via free radicals and ROS due to the glutathione level and antioxidant enzyme depletion [24, 25]. In recent years, there has been an attempt to use extracts from medicinal plants with antioxidant properties to treat different diseases [19, 26]. Based on previous studies, CS is one of the herbs with antioxidant and free-radical scavenger activities that can be used in renal damage and oxidative injury [11, 19]. In the current study, the effect of CS on cisplatin-induced nephrotoxicity was investigated using experimental rat models.

Suzuki et al. investigated the effects of CS on diabetic nephropathy induced by streptozotocin (STZ). Based on their results, the increase in creatinine clearance in the administered group (STZ+ CS) was suggestively inhibited compared with the non-treated group (STZ). Such a trend was also observed in urinary albumin excretion. The latter results showed that the aquatic extract of CS prevented the glomerular hyperfiltration. Accordingly, the water extract of CS suppressed the development of diabetic glomerular sclerosis in STZ-induced diabetic rat [27].Sepehri et al. in their experimental study evaluated the effect of CS against gentamicin (GM)-induced nephrotoxicity. They used different doses of CS (200, 300, 400 and 500 mg/kg). As per their results, plasma creatinine and urea levels significantly increased in the GM group. CS management (200 and 300 mg/kg) with GM injection significantly reduced the serum creatinine, but not urea levels compared with the GM group. Acute tubular necrosis hyaline casts in the tubular lumen, interstitial nephritis, and glomerular variations were observed in the GM group, using histopathological techniques. Notably, co-treatment of CS with GM considerably decreased the interstitial nephritis. Furthermore, high doses of CS caused hyaline cast formation, apoptosis, congestion, and swelling of the renal tubules. Consequently, CS might improve nephropathy during prolonged therapeutic use of GM and related aminoglycosides [19].In the present study, we demonstrated that daily CS administration significantly improved the cisplatin toxicity to the kidney and liver, as confirmed by biochemical and microscopic examination. Cisplatin administration showed a significant increase in the urea levels compared to normal rats, which indicated renal failure.Based on our results, cisplatin administration caused severe degeneration in the glomeruli and both proximal and distal tubules. Functional nephrotoxicity indices, such as the BUN, were increased in the cisplatin-injected animals compared to the control group. Furthermore, cisplatin administration caused severe damage in the liver, as assessed microscopically. The CS extract with 200 mg dose protected the elevation of the cisplatin-induced BUN. The histopathological evaluation also showed that pre-treatment with CS caused a significant reduction in the tubular necrosis compared to cisplatin alone which is in the same line with Ingale et al. report [4].

Although the exact mechanism of nephrotoxicity induced by cisplatin is not well understood [28], previous reports have suggested that the free radicals and ROS are involved due to the depletion of glutathione concentration and antioxidant enzyme activities in the kidney [29, 30]. The above observations support the mechanism of nephrotoxicity induced by cisplatin in animals, which is partially related to the depletion of the renal antioxidant system. Treatment of cisplatin-induced nephrotoxicity with CS could significantly prevent the reduction of these renal antioxidant systems. The quantitative stereological evaluation was not performed, which was one of the limitations of this study.

Conclusion

CS has the potential to attenuate nephrotoxicity and lipid peroxidation induced by cisplatin in rats.

Acknowledgment

This research was founded by Grant no.6905 Shiraz University of Medical Scieneses.

Conflict of Interest

All authors declare there is no any conflict of interest in presenting the current work.

Figure 1. Effects of CS on lipid peroxidation levels (MDA) in rats. Data are presented as mean ±SD. *P<0.01 vs. control group; **P<0.01 vs. cisplatin-treated group

Figure 2. Effects of CS on serum levels of the BUN in rats. Data are presented as mean ± SD. *P<0.01 vs. control group; **P<0.01 vs. cisplatin-treated group

Figure 4. Histological sections of the kidney (H&E staining, ×200) in experimental rats. A: Group C1 showing adequate tissue with no specific pathologic change; B: Group C2 showing extensive tubular necrosis and desquamation as well as signs of regeneration magnified in lower left of the image; C: Group CS showing minimal damage, focal loss of brush borders, and very few hyaline casts; D: Group CS1showing moderate tubular necrosis and cast formation admixed with normal tubules; E: Group CS2 showing minimal tubular necrosis.

Figure 3. Effects of CS on AST (A) and ALT (B). Data are presented as mean ± SD. *P<0.01 vs. control group; **P<0.01 vs. cisplatin-treated group

References

1. Ciarimboli G. Membrane transporters as mediators of cisplatin side-effects. Anticancer Res. 2014;34(1):547-50.
2. Ozkok A, Edelstein CL. Pathophysiology of cisplatin-induced acute kidney injury. Biomed Res Int. vol. 2014, Article ID 967826, 17 pages, 2014. 2014;2014.
3. Razzaque MS. Cisplatin nephropathy: is cytotoxicity avoidable? Nephrol Dial Transplant. 2007;22(8):2112-6.
4. Ingale KG, Thakurdesai PA, Vyawahare NS. Protective effect of Hygrophila spinosa against cisplatin induced nephrotoxicity in rats. Indian J Pharmacol. 2013;45(3):232.
5. Chirino YI, Pedraza-Chaverri J. Role of oxidative and nitrosative stress in cisplatin-induced nephrotoxicity. Exp Toxicol Pathol. 2009;61(3):223-42.
6. Zhang M, Ning G, Shou C, Lu Y, Hong D, Zheng X. Inhibitory effect of jujuboside A on glutamate-mediated excitatory signal pathway in hippocampus. Planta Med. 2003;69(08):692-5.
7. Cummings BS, Schnellmann RG. Cisplatin-induced renal cell apoptosis: caspase 3-dependent and-independent pathways. J Pharmacol Exp Ther. 2002;302(1):8-17.
8. Parlakpinar H, Tasdemir S, Polat A, Bay-Karabulut A, Vardi N, Ucar M, et al. Protective role of caffeic acid phenethyl ester (cape) on gentamicin-induced acute renal toxicity in rats. Toxicology. 2005;207(2):169-77.
9. Mohan IK, Khan M, Shobha JC, Naidu MUR, Prayag A, Kuppusamy P, et al. Protection against cisplatin-induced nephrotoxicity by Spirulina in rats. Cancer Chemother Pharmacol. 2006;58(6):802-8.
10. Arunkumar P, Viswanatha G, Radheshyam N, Mukund H, Belliyappa M. Science behind cisplatin-induced nephrotoxicity in humans: A clinical study. Asian Pac J Trop Biomed. 2012;2(8):640-4.
11. El-Ghorab A, El-Massry KF, Shibamoto T. Chemical composition of the volatile extract and antioxidant activities of the volatile and nonvolatile extracts of Egyptian corn silk (Zea mays L.). J Agric Food Chem. 2007;55(22):9124-7.
12. Ebrahimzadeh MA, Pourmorad F, Hafezi S. Antioxidant activities of Iranian corn silk. Turk J Biol. 2008;32(1):43-9.
13. Kim SR, Ha AW, Choi HJ, Kim SL, Kang HJ, Kim MH, et al. Corn silk extract improves benign prostatic hyperplasia in experimental rat model. Nutr Res Pract. 2017;11(5):373-80.
14. Adedapo A, Babarinsa O, Oyagbemi A, Adedapo A, Omobowale T. Cardiotoxicity study of the aqueous extract of corn silk in rats. Mac Vet Rev. 2016;39(1):43-9.
15. Hasanudin K, Hashim P, Mustafa S. Corn silk (Stigma maydis) in healthcare: a phytochemical and pharmacological review. Molecules. 2012;17(8):9697-715.
16. Zheng L-L, Wen G, Yuan M-Y, Gao F. Ultrasound-assisted extraction of total flavonoids from corn silk and their antioxidant activity. J Chem. vol. 2016, Article ID 8768130, 5 pages, 2016.2016;2016.
17. Liu J, Wang C, Wang Z, Zhang C, Lu S, Liu J. The antioxidant and free-radical scavenging activities of extract and fractions from corn silk (Zea mays L.) and related flavone glycosides. Food Chem. 2011;126(1):261-9.
18. Wang C, Zhang T, Liu J, Lu S, Zhang C, Wang E, et al. Subchronic toxicity study of corn silk with rats. J ethnopharmacol. 2011;137(1):36-43.
19. Sepehri G, Derakhshanfar A, Zadeh FY. Protective effects of corn silk extract administration on gentamicin-induced nephrotoxicity in rat. Comp Clin Path. 2011;20(1):89-94.
20. Karami M, Shokerzadeh M, Naghshvar F, Ala S, Fezbakhsh R, Nosrati A, et al. The Renal Protective Effects of Corn Silk and Feijoa by using in situ Rat Renal System. Iranian J Toxicol. 2014;8(25):1060-7.
21. Moghadam MG, Ansari I, Roghani M, Ghanem A, Mehdizade N. The Effect of Oral Administration of Hypericum Perforatum on Serum Glucose and Lipids, Hepatic Enzymes and Lipid Peroxidation in Streptozotocin-Induced Diabetic Rats. Galen. 2017;6(4):319-29.
22. Movassaghi S, Oudarji AY, Sharifi ZN. Effect of Pentoxifylline on Apoptosis of Kidney’s Cells Following Acute Methamphetamine Administration in Male Wistar Rats. Galen. 2016;5(3):131-38.
23. İşeri S, Ercan F, Gedik N, Yüksel M, Alican I. Simvastatin attenuates cisplatin-induced kidney and liver damage in rats. Toxicology. 2007;230(2-3):256-64.
24. Sherif IO. Amelioration of cisplatin-induced nephrotoxicity in rats by triterpenoid saponin of Terminalia arjuna. Clin Exp Nephrol. 2015;19(4):591-7.
25. Palipoch S, Punsawad C, Chinnapun D, Suwannalert P. Amelioration of cisplatin-induced nephrotoxicity in rats by curcumin and α-tocopherol. Trop J Pharm Res. 2013;12(6):973-9.
26. Ajith T, Nivitha V, Usha S. Zingiber officinale Roscoe alone and in combination with α-tocopherol protect the kidney against cisplatin-induced acute renal failure. Food Chem Toxicol. 2007;45(6):921-7.
27. Suzuki R, Okada Y, Okuyama T. The favorable effect of style of Zea mays L. on streptozotocin induced diabetic nephropathy. Biol Pharm Bull. 2005;28(5):919-20.
28. Dasari S, Tchounwou PB. Cisplatin in cancer therapy: molecular mechanisms of action. Eur J Pharmacol. 2014;740:364-78.
29. Ajith T, Jose N, Janardhanan K. Amelioration of cisplatin induced nephrotoxicity in mice by ethyl acetate extract of a polypore fungus, Phellinus rimosus. J Exp Clin Cancer Res: CR. 2002;21(2):213-7.
30. Çetin R, Devrim E, Kılıçoğlu B, Avcı A, Çandır Ö, Durak İ. Cisplatin impairs antioxidant system and causes oxidation in rat kidney tissues: possible protective roles of natural antioxidant foods. J Appl Toxicol. 2006;26(1):42-6.