CC BY
1
JOURNAL OF CLINICAL MEDICINE OF KAZAKHSTAN
Озык, Макала / Оригинальная Статья / Original Article DOI: 110.23950/1812-2892-JCMK-00563
Hematological dynamics following the co-administration of Resveratrol and Cisplatin in Sprague-Dawley rats
Izuchukwu Azuka Okafor1, Johnson Okwudili Nweke2, Uchenna Somtochukwu Nnamah3
Abstract
Background: Cisplatin, a platinum-based chemotherapy agent, is used in the effective treatment of a wide range of malignant cancers. Resveratrol, a polyphenolic plant-derived compound, has been shown to have several biological effects including protecting the body against several forms of damages.
Objective: This study assessed the effects of cisplatin and supplementation with resveratrol on some hematological parameters in Sprague-Dawley rats.
Material and Methods: Forty-five adult female Sprague-Dawley rats, grouped into 9, were used for this experimental study. Group 1 served as the control group and received distilled water only. Groups 2 and 9 received Cisplatin only while groups 3, 4, and 5 received different doses of Resveratrol after a single dose of Cisplatin. Groups 6, 7, and 8 received Resveratrol before Cisplatin.
Results: At the end of administration, blood samples were collected and different hematological parameters were accessed. Groups 2 and 5 showed a significantly higher Total WBC when compared to the control group (p<0.05). Group 7 showed a significantly higher (p<0.05) neutrophil count compared to both the control group and group 2, but a significant decrease in the lymphocyte count compared to the same groups (p<0.05). Groups 6 and 7 showed a significant reduction in monocyte when compared to the control group (p<0.05). There was no significant difference in the eosinophil count of all treatment groups when compared to the control group and cisplatin groups (p<0.05). Basophil increased significantly in group 7 when compared to the control group and group 2 (p<0.05). More so, a significant decrease in the Haemoglobin (Hb), Packed cell volume (PCV) and Mean corpuscular volume (MCV) level were observed when compared to group 9 (p<0.05).
Conclusion: This study showed that cisplatin-induced alteration in some hematological parameters is reversible by treatment with resveratrol. More so, resveratrol supplementation should be incorporated as part of dietary nutrients for patients on cisplatin during chemotherapy.
Keywords: cisplatin, hematology, resveratrol, Sprague-Dawley, supplementation
РЕСВЕРАТРОЛ МЕН ЦИСПЛАТИНД1 ЕНГ1ЗГЕННЕН КЕЙ1Н СПРЕГ-ДОУЛИ ЕГЕУЦ¥ЙРЬЩТАРЫНЬЩ ГЕМАТОЛОГИЯЛЬЩ ПАРАМЕТРЛЕР1
Изучукву Азука Окафор1, Джонсон Оквудили Нвеке2, Ученна Сомточукву Ннамах3
'Анатомия кафедрасы, HerÍ3ri медицинальщ гылымдар факультету Денсаульщ гылымы колледжу Ннамди Азикиве Университету Нневи, Анамбра штаты, Нигерия 2Медицинальщ-зертханальщ гылымдар кафедрасы, Iykenson Medical and Diagnostic Co. Ltd., Аква, Анамбра штаты, Нигерия зМедициналыщ-зертханалыщ гылымдар кафедрасы, Ннамди Азикиве университетшщ ауруханасы, Нневи, Анамбра штаты, Нигерия
Т¥ЖЫРЫМДАМА
Kipicne: Цисплатин - бул TYрлi |^атерл1 iсiктердi ™ii4ai емдеуде пайдаланылатын химиотерапевтикалык препараты курайтын пла-тин. Ресвератрол, шыгуы еамдк TYрiнен болатын полифенольд байланысудыц закымдалудыц ^6ip формаларынан организмдi коргауды косканда, бiрнеше биологиялык эсерi бар екендИ дэлелдендк
Максаты: Осы зерттеуде Спрег-Доули егеукуйрыктарындагы кейбiр гематологиялык параметрлерге ресвератролдыц коспалары жэне цисплатиннщ эсерi багаланды.
Материалдар мен эдicтep: Осы зерттеуде 9 топка белнен кырык бес ересек Спрег-Доули егеукуйрыктары колданылды. 1-топ бакылау тобы болды жэне дистилденген суды гана кабылдады. 2-шл жэне 9-шы топтар тек кана цисплатин кабылдады, сол уакытта 3, 4-шл
'Department of Anatomy, Faculty of Basic Medical Sciences, College of Health Sciences, Nnamdi Azikiwe University, Nnewi, Anambra State, Nigeria Department of Medical Laboratory Science, Iykenson Medical and Diagnostic Co. Ltd., Awka, Anambra State, Nigeria
Department of Medical Laboratory Science, Nnamdi Azikiwe University Teaching Hospital, Nnewi, Anambra State, Nigeria
© ®
This work is licensed under a Creative Commons Attribution 4.0 International License
Received: 2018-04-17 Accepted: 2018-05-03 UDC: 616.1
J Clin Med Kaz 2018;2(48):22-27
Corresponding Author: Izuchukwu Azuka Okafor, Department of Anatomy, Faculty of Basic Medical Sciences, College of Health Sciences, Nnamdi Azikiwe University, Nnewi Campus P.M.B. 5001, Nnewi, Anambra State, Nigeria. Tel.: +2348030716546
E-mail: okaforizuchukwu33@gmail.com
жэне 5-шл топтар цисплатиннщ бiрiншi дозасынан кейiн ресвератролдыц TYрлi дозаларын кабылдады. 6, 7-шi жэне 8-шi топтар цисплатиннщ алдынан ресвератролды кабылдады.
Нэтижелерк Препараттарды енгiзудi аяктау бойынша канныц Yлгiлерiн жинадык жэне TYрлi гематологиялык параметрлерiн багаладык. 2-шi жэне 5-шл топтарда лейкоциттердщ жалпы мвлшерi бакылау тобымен (р<0.05) салыстырганда айтарлыктай жогары. 7-шi топта бакылау тобына жэне 2-шл топка караганда, нейтрофилдердщ мвлшерi айтарлыктай жогары (р<0.05), алайда сол топтармен салыстырганда лимфоциттердщ мвлшершщ айтарлыктай твмендеуi байкалды (р<0.05). 6-шы жэне 7-шл топтар бакылау тобымен салыстырганда (р<0.05), моноциттер мвлшершщ айтарлыктай твмендИн кврсеттi.
Барлык зерттелетiн топтарда эозинофилдердщ мвлшерiндегi айтарлыктай ерекшелiгi бакылау топтармен жэне цисплатиндi (р<0.05) кабылдаган топтармен салыстырганда байкалмады. Базофилдердщ мвлшерi бакылау тобымен жэне 2-шл топпен салыстырганда (р<0.05) 7-шi топтарда айтарлыктай артты. 9-шы топпен салыстырганда, гемоглобин (НЬ), гематокриттi саны (РСУ) мен эритроциттщ орташа квлемi (МСУ) децгешнщ айтарлыктай твмендеуi байкалды.
Корытынды: Осы зерттеу цисплатиндi енгiзумен туындаган гематологиялык параметрлершщ взгеруi - ресвератролмен терапиясыньщ квмепмен кайтымды екендiгiн кврсеттi. Ресвератролдьщ коспасын химиотерапия кезiнде цисплатинге диеталык кунарлы заттектердщ бiр бвлiгi ретiнде косу кажет.
Негiзгi свздер: цисплатин, гематология, ресвератрол, Спрег-Доули, коспа
ГЕМАТОЛОГИЧЕСКИЕ ПАРАМЕТРЫ У КРЫС СПРЕГ-ДОУЛИ ПОСЛЕ ВВЕДЕНИЯ РЕСВЕРАТРОЛА И ЦИСПЛАТИНА
Изучукву Азука Окафор1, Джонсон Оквудили Нвеке2, Ученна Сомточукву Ннамах3
'Кафедра анатомии, Факультет основных медицинских наук, Колледж наук здоровья, Университет Ннамди Азикиве, Нневи, Штат Анамбра, Нигерия 2Кафедра медицинско-лабораторных наук, Iykenson Medical and Diagnostic Co. Ltd., Аква, Штат Анамбра, Нигерия 3Кафедра медицинско-лабораторных наук, Больница университета Ннамди Азикиве, Нневи, Штат Анамбра, Нигерия
РЕЗЮМЕ
Введение: Цисплатин это платин, содержащий химиотерапевтический препарат, используемый для эффективного лечения различных злокачественных опухолей. Было доказано, что ресвератрол, полифенольное соединение растительного происхождения, имеет несколько биологических эффектов, включая защиту организма от некоторых форм поражений.
Цель: В данном исследовании оценивались эффекты цисплатина и добавки ресвератрола на некоторые гематологические параметры у крыс Спрег-Доули.
Материалы и методы: В данном исследовании было использовано сорок пять взрослых крыс Спрег-Доули, разделенных на 9 групп. Группа 1 стала контрольной группой и получала только дистиллированную воду. Группы 2 и 9 получали только цисплатин, в то врем как группы 3, 4 и 5 получали различные дозы ресвератрола после одной дозы цисплатина. Группы 6, 7 и 8 получали ресвератрол перед цисплатином.
Результаты: По завершению введения препаратов, собрали образцы крови и оценили различные гематологические параметры. У групп 2 и 5 общее количество лейкоцитов было значительно выше, по сравнению с контрольной группой (p<0.05). В группе 7 количество нейтрофилов было значительно больше (p<0.05), чем в контрольной группе и группе 2, но отмечалось значительное снижение количества лимфоцитов, по сравнению с теми же группами (p<0.05). Группы 6 и 7 показали значительное снижение количества моноцитов, по сравнению с контрольной группой (p<0.05). Значительной разницы в количестве эозинофилов во всех исследуемых группах не отмечено, по сравнению с контрольной группой и группами, получавшими цисплатин (p<0.05). Количество базофилов значительно увеличилось в группе 7, по сравнению с контрольной группой и группой 2 (p<0.05). Более того, наблюдалось значительное снижение уровня гемоглобина (Hb), гематокритного числа (PCV) и среднего объема эритроцита (MCV), по сравнению с группой 9 (p<0.05).
Заключение: Данное исследование показало, что изменение гематологических параметров, вызванное введением цисплатина, обратимо с помощью терапии ресвератролом. Более того, добавку ресвератрола необходимо включать как часть диетических питательных веществ к цисплатину во время химиотерапии.
Ключевые слова: цисплатин, гематология, ресвератрол, Спрег-Доули, добавка
Introduction
Cisplatin is an effective chemotherapeutic agent, which belongs in the platinum-based anti-neoplastic family of medications [1]. It is used in the treatment of a wide range of malignant diseases including testicular cancer, ovarian cancer, cervical cancer, breast cancer, bladder cancer, head and neck cancer, esophageal cancer, lung cancer, mesothelioma, brain tumors and neuroblastoma. However, it exhibits certain toxic effects on several organs such as the kidneys and liver, which interfere with its therapeutic efficiency [2]. Previous works by Wood and Hrusheeky in 1995 [3] showed, that cisplatin causes significant effects on various hematological parameters only during chronic treatment in humans and rats. Cisplatin-induced anemia is also a well-known side-effect [4], which occurs in 9-40% of patients [5]. However, some previous results showed that high, acute doses of cisplatin did not affect the RBC maturation in rats [1].
Resveratrol (3, 4' 5-trihydroxystilbene) is a polyphenol synthesized by a wide variety of plant species, including aliments such as grapes, peanuts and mulberries, in response to injury, UV irradiation and fungal attack. Resveratrol belongs to a group of compounds called polyphenols; it is the most extensively studied and has long been considered a therapeutic agent for various diseases, including inflammatory diseases [7]. Studies by Loehrer et al., 1998 [8] also showed resveratrol to
have cytotoxic effects against human cancer cell lines derived from various tumor types. Preclinical studies have revealed this compound to have several biological effects including antioxidant effects thus protect the body against damages.
Thus, this study investigated the effects of cisplatin and supplementation with resveratrol on various hematological parameters in Sprague-Dawley rats.
Material and methods Animal Care and Handling
This experimental animal study was carried out in the research laboratory of the Department of Anatomy, College of Medicine of the University of Lagos, Nigeria. Forty-five adult female Sprague-Dawley rats with average weight of 160 g were procured from the Animal House, College of Medicine of the University of Lagos. They were acclimatized for two weeks to exclude any intercurrent infection under standard housing of 24 ± 2°C and 12 hr light/dark cycle. The rats were fed with standard rat chow and water ad-libitum.
Experimental Drugs
Resveratrol with brand name 'Restorlyf', manufactured by Nature's Way U.S.A., was procured from Alliance in Motion Global Ltd., Ikeja, Lagos, Nigeria. 325 mg of Resveratrol were diluted immediately before each use in 20 ml of distilled water
and doses of 5, 10 and 20 mg/kg/b.wt were administered orally using the oral cannula. The remaining formulation was discarded after each use. The drug dosages and formulations were chosen on the basis of previously published studies on Resveratrol [7, 8]. Cisplatin (Zuplatin, 50 mg/ 50 ml) injection manufactured by Taj pharmaceuticals Ltd. India was procured from Bayston Pharmacy, Mushin, Lagos, Nigeria. The injection was given intraperitoneally according to body weight in a single dose of 5 mg/kg. The drug dosages were chosen on the basis of previously published studies on Cisplatin [9, 10].
Experimental Design
Forty-five adult female Sprague-Dawley rats with average weight of 160 g were divided into 9 groups (n=5) and used for this experimental study. Group 1 served as the normal control group and received distilled water only. Group 2 was given only a single dose intra-peritoneal injection of 5 mg/kg b.wt Cisplatin and allowed to stay for 7 days before sacrifice. Groups 3, 4 and 5 were given 5, 10 and 20 mg/kg/b.wt. Resveratrol respectively for 7 days, starting 24 hours after a single dose intra-peritoneal injection of 5 mg/kg/b.wt. Groups 6, 7 and 8 were treated with 5, 10 and 20 mg/kg/b.wt. Resveratrol respectively for 14 days before a single dose intra-peritoneal injection of 5 mg/kg b.wt. Cisplatin; the respective doses of Resveratrol treatment was repeated for another 7 days. Group 9 was given only a single dose intra-peritoneal injection of 5 mg/kg b.wt. Cisplatin and allowed to stay for 21 days before sacrifice. At the end of the study, body weight and hematological indices were taken.
Animal Sacrifice and Sample Collection
At the end of the study, the animals were fasted overnight and sacrificed by cervical dislocation. Blood samples were collected by cardiac. The serum was separated by allowing blood sample to stand for 15 minutes at 25 oC and then centrifuged at 4000 rpm for 20 minutes. Serum was kept in plastic vials at -40oC until further biochemical analysis.
Determination of Hematological Parameters
Full blood count was done using BC-5300 Mindray Autohematology analyzer and all examinations were done using the CDC laboratory manual guide 2016. The blood parameters analysed include red blood cell count (RBC), white blood cell (WBC) and differentials' count, haemoglobin (Hb) concentration, packed cell volume (PCV). Mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and
mean corpuscular haemoglobin concentration (MCHC) were also computed according to Jain (1986).
Ethical consideration
Ethical approval was gotten from the College of Medicine of the University of Lagos Health Research Ethics Committee (CMULHREC) with ID number CMULHREC/09/16/025. All procedures were carried out in accordance with the National Academy of Science's Guide for Care and Use of Laboratory Animals [11].
Statistical analysis
The results were analyzed using the Statistical Package for the Social Sciences version 21.0 (SPSS, Chicago, IL, USA). Data was reported as mean ± SD Differences between mean and the main effects of treatment group were determined by the one way analysis of variance (ANOVA) and multiple comparisons was done using the LSD post-hoc tests. The mean difference is significant at the 0.05 level (P<0.05).
Results
White blood cell counts and Differentials (Table I)
Groups 2 and 5 showed a significantly higher Total WBC when compared to the control group (p<0.05). The control group and groups 6 - 9 showed a significant reduction in Total WBC when compared to the group 2 (p<0.05). All treatment groups showed no significant difference in their Neutrophil differential when compared to the control group, groups 2 and 9 (p > 0.05) except for group 7 that have a significantly higher neutrophil count compared to both the control group and group 2 (p<0.05). Group 7 showed a significant decrease in the lymphocyte count compared to the control group (p<0.05). The control group and groups 2, 4, 6 and 8 showed a significant increase in lymphocyte count when compared to the group 9 (p<0.05). Only group 7 showed a significant decrease in lymphocyte when compared to the control group and group 2. Groups 6 and 7 showed a significant reduction in monocyte when compared to the control group (p<0.05).
There is no significant difference in the eosinophil count of all treatment groups when compared to the control group and groups 2 and 9 (p>0.05). Basophil increased significantly in group 7 when compared to the control group and the group 2 (p<0.05).
Table 1
The Effect of Cisplatin and Supplementation with Resveratrol on the White blood cells and Differentials.
GROUPS TOTAL WBC (/103cm3) NEUTROPHIL (%) LYMPHOCYTE (%) MONOCYTE (%) EOSINOPHIL (%) BASOPHIL (%)
GROUP 1 6.57 ± 1.50 b 31.67 ± 1.67 64.67 ± 1.33 c 2.67 ± 0.33 0.67 ± 0.33 0.33 ± 0.33
GROUP 2 11.70 ± 2.34a,c 31.33 ± 2.33 65.67 ± 2.73 c 2.00 ± 0.00 0.67 ± 0.33 0.33 ± 0.33
GROUP 3 8.00 ± 1.04 33.67 ± 2.73 63.67± 3.33 2.00 ± 0.58 0.67 ± 0.33 0.00 ± 0.00
GROUP 4 8.90 ± 1.25 c 31.00 ± 4.62 65.67 ± 4.63 c 1.67 ± 0.67 1.33 ± 0.67 0.33 ± 0.33
GROUP 5 11.67 ± 1.66a,c 33.67 ± 0.97 63.00 ± 2.89 2.00 ± 0.58 1.33 ± 0.33 0.00 ± 0 .00
GROUP 6 4.63 ± 0.15 b 32.67 ± 2.03 65.33 ± 2.03 c 1.00 ± 0.00 a 0.67 ± 0.33 0.33 ± 0.33
GROUP 7 5.9667 ± 1.72 b 46.00 ± 5.69a,b 50.33 ± 6.36a,b 1.00 ± 0.58 a 1.67 ± 1.20 1.33 ± 0.33a,b
GROUP 8 5.03 ± 1.28 b 32.33 ± 4.18 64.67 ± 4.18 c 1.67 ± 0.33 0.67 ± 0.33 0.67 ± 0.33
GROUP 9 4.27 ± .769 b 40.67 ± 0.89 53.67 ± 2.85 2.00 ± 0.58 3.00 ± 2.00 0.67 ± 0.33
Values are expressed as mean ± Standard Error of Mean (SEM). ap<0.05 significant compared to control group; bp<0.05 significant compared to group 2; cp<0.05 significant compared to group 9. WBC means white blood cells. CIS = cisplatin; RES = Resveratrol; MED = Medium. Group 1 = control; Group 2 = Cisplatin only; Group 3 = CIS + RES LOW; Group 4 = CIS + RES MED; Group 5 = CIS + RES HIGH; Group 6 = RES LOW + CIS + RES LOW; Group 7 = RES MED + CIS + RES MED; Group 8 = RES HIGH + CIS + RES HIGH; Group 9 = Cisplatin only
Anaemia Blood markers (Table II)
The Red Blood Count (RBC) count was not significantly different in all the treatment groups when compared to the control group and cisplatin groups (p>0.05). All the experimental groups showed no significant difference in the Hemoglobin (Hb) levels and packed cell volume (PCV) when compared to groups 1, 2 and 9 except group 4 which showed a significant decrease when compared to group 9 (p<0.05). There was no difference in the
Mean Corpuscular Volume MCV of all treatment groups when compared with the control group and cisplatin groups (p>0.05) except for groups 4 and 5 which showed a significant decrease when compared with group 9 (p<0.05). There was no significant difference in the Mean Corpuscular Hemoglobin (MCH) and Mean Corpuscular Hemoglobin Concentration (MCHC) between all experimental groups (p>0.05).
Table 2
The Effect of Cisplatin and Supplementation with Resveratrol on RBC, Hb, PCV and their derivatives in female Sprague-Dawley rats.
Values are expressed as mean ± Standard Error of Mean (SEM). ap<0.05 significant compared to control group; bp<0.05 significant compared to group 2; cp<0.05 significant compared to group 9. CIS = cisplatin; RES = Resveratrol; MED = Medium. Group 1 = control; Group 2 = Cisplatin only; Group 3 = CIS + RES LOW; Group 4 = CIS + RES MED; Group 5 = CIS + RES HIGH; Group 6 = RES LOW + CIS + RES LOW; Group 7 = RES MED + CIS + RES MED; Group 8 = RES HIGH + CIS + RES HIGH; Group 9 = Cisplatin only. MCV means mean corpuscular volume; MCH means mean corpuscular hemoglobin; MCHC means mean corpuscular hemoglobin concentration.
GROUPS Red Blood Cell Hemoglobin Packed Cell MCV MCH MCHC
(/cm3) (g/dl) Volume (%) (Fl) (Pg) (g/dl)
GROUP 1 6.73 ± 0.35 12.63 ± 0 .44 40.67 ± 2.03 60.33 ± 0.33 18.67 ± 0.33 31.33±0.33
GROUP 2 6.87 ± 0.19 12.90 ± 0.21 40.67 ± 0 .88 58.67 ± 0.33 18.67 ± 0.33 32.33±0.33
GROUP 3 7.03 ± 0.09 13.47 ± 0.12 42.00 ± 1.15 59.00 ± 0.58 19.33 ± 0.33 32.33±0.67
GROUP 4 6.17 ± 0 .46 11.80 ± 0.91 c 35.67 ± 2 .91 c 57.67 ±0.88c 19.00 ±0.00 33.00±0.58
GROUP 5 6.83 ± 0.09 12.50 ± 0.25 38.33 ± 1.33 56.67 ±1.67c 18.33 ± 0.33 32.33±0.67
GROUP 6 6.37 ± 0.20 12.50 ± 0.40 39.00 ± 2.00 61.67 ± 2.03 19.67 ± 0.33 32.00±0.58
GROUP 7 6.77 ± 0.27 12.87 ± 0.38 42.00 ± 2.51 62.00 ± 3.06 18.67 ± 0.33 31.00±1.15
GROUP 8 6.63 ± 0.33 12.70 ± 0.50 39.67 ± 1.86 60.00 ± 0.00 19.00 ± 0.00 31.67±0.33
GROUP 9 6.90 ± 0.10 13.20± 0 .21 43.33 ± 1.20 62.33 ± 0.33 19.00 ± 0.00 30.67±0.33
Platelet count (Table III)
There is no significant difference in the platelet count of all experimental groups when compared with group 1 except for group 2 which showed a significant increase (p<0.05). All experimental groups showed significant decrease in platelet
count when compared with group 2 (p<0.05) except for group 8 which showed no significant difference (p>0.05). Only group 4 showed a significant decrease in platelet count when compared with group 9 (p<0.05).
The Effect of Cisplatin and Supplementation with Resveratrol on the Platelet count of female Sprague-Dawley rats.
GROUPS PLATELET
(/103cm3)
GROUP 1 0.73± 0.03 b
GROUP 2 1.03 ± 0.11 a,c
GROUP 3 0.68 ± 0.05 b
GROUP 4 0.37 ± 0.05b,c
GROUP 5 0.65 ± 0.09b
GROUP 6 0.63 ± 0.05 b
GROUP 7 0.68 ± 0.10 b
GROUP 8 0.83 ± 0.13
GROUP 9 0.74 ± 0.10 b
Values are expressed as mean ± Standard Error of Mean (SEM). ap<0.05 significant compared to control group; bp<0.05 significant compared to group 2; cp<0.05 significant compared to group 9. CIS = cisplatin; RES = Resveratrol; MED = Medium. Group 1 = normal control; Group 2 = Cisplatin only; Group 3 = CIS + RES LOW; Group 4 = CIS + RES MED; Group 5 = CIS + RES HIGH; Group 6 = RES LOW + CIS + RES LOW; Group 7 = RES MED + CIS + RES MED; Group 8 = RES HIGH + CIS + RES HIGH; Group 9 = Cisplatin only.
Discussion
This present study assessed the effect of cisplatin and supplementation with resveratrol on some hematological parameters including white blood cell counts, differentials, anaemia blood markers and platelet counts in Sprague- Dawley rats. The significant differences observed in the various Parameters were compared with the normal control and the cisplatin control groups.
Research findings have established that haemolytic anaemia can be developed after several courses of cisplatin which was suggested to be due to the reaction of an antibody with a cisplatin-red-cell membrane [12]. Thus, anaemia has been stated as a common side effect of cisplatin, especially after repeated infusions; the primary mechanism is a myelosuppression caused by cisplatin's interference with iron metabolism, resulting in a lower count of red cell precursors [13]. Some authors report that haemolysis is caused by an antiglobulin antibody directed against red cell membrane-bound cisplatin. Cisplatin- based therapy has been discovered
to result in a cumulative anaemia that is disproportionate to the effects on other blood cells. On examination of the hematological parameters in this present study, we found no significant change between groups in the red blood cell, eosinophils and mean corpuscular haemoglobin concentration (MCHC) (p>0.05).
The total white blood cell count showed the cisplatin control group to have significantly increased when compared to the normal control group (p<0.05). The other treatment groups were observed to be the same with the normal group except for rats post-treated with high dose of Resveratrol which showed a significant decrease in total white blood cell count.
Unlike Juan and co-workers in 2002 who found out that high-dose resveratrol administration did not change hematological parameters in rats, this present study found WBC count to be significantly lower in the group that received high dose of resveratrol, as compared to the control group. This decrease could be as a result of the anti-inflammatory effect of resveratrol, which is in consonance with the observation of Donnelly et al., 2004 [14]. This is equally in line with the observation of Hi§miogullan et al., 2013 [6] who reported that WBC count was decreased in rats given resveratrol.
An assessment of the white blood cell (WBC) differentials showed that there was no significant change in the neutrophil of all groups except for rats treated with prophylactic medium dose of Resveratrol which showed a significant increase. An increase in leucocyte number could be consequence of infection and inflammation during cisplatin treatment and cisplatin metabolism in experimental rats [15]. This same group equally showed a significant decrease in the lymphocyte compared to the normal group as opposed to the no significant change seen across the other groups. These other groups also showed a significant increase in lymphocytes when compared to the cisplatin control group. Thus, lymphopenia which is often found in patients undergoing chemotherapy and radiotherapy was seen to be prevented across the resveratrol -treated groups.
The low and medium dose prophylactic groups showed a decrease in their number of monocyte while only the medium dose group showed a significant increase in basophil when compared to the normal control groups (p<0.05). This increase in basophils could have been as a result of an inflammation, as basophils take part in inflammatory response [16].
Our study, on assessment of the anemia blood markers showed that haemoglobin (Hb) and packed cell volume (PCV) of all treatment groups is the same with the normal control group (p>0.05). However, a decrease was observed in the haemoglobin level and packed cell volume of the group post-treated with medium dose Resveratrol when compared with the cisplatin control group. Mean cell volume (MCV) was seen to be significantly decreased in the medium and high dose post-treated with resveratrol when compared to the cisplatin
control group, but other treatment groups showed no significant difference in MCV. There was no significant change observed in all groups when compared to the normal control in the mean cell haemoglobin (MCH) of the treated rats. However, only the medium dose post-treatment with Resveratrol showed a significant increase in MCH when compared to the normal control (p<0.05). There were no significant differences in the RBC count and MCHC in all treatment groups when compared to the control group and cisplatin groups.
Thus, the anemia blood markers were seen to be normal and unaffected in the cisplatin control groups, indicating that the cisplatin-treated groups did not suffer anaemia. This could have been because there were no chronic or repeated infusions of cisplatin on the rats. This finding is in agreement with previous research results by Markovic et al., 2010 [1] which showed that high, acute doses of cisplatin did not affect the RBC maturation in rats. More so, another study, in 1995 [3], showed that cisplatin causes significant effects on hematological parameters only during chronic treatment in humans and rats [4].
Previous studies by Olas and his colleagues in 2005 [15] have shown that cisplatin causes oxidative stress in human platelets and lymphocytes, which might reflect on their life expectancy, the induction of apoptosis, and thereby ultimately reduce the number of these cells in the blood. However, the platelet assessment in this study rather showed a significant increase in platelet count of cisplatin control group when compared to the normal group (p<0.05). This change seen may be unconnected to the cisplatin-triggered platelet activation as discovered in research by Calvert et al., 1989 [17]. Also, there was no significant difference in the platelet count across all the experimental groups when compared to the control group (p<0.05).
Conclusion
Cisplatin used in the treatment of a wide range of malignant diseases has been associated with notable toxic effects on several organs. In this study, we have been able to show that cisplatin-induced alteration in some hematological parameters is reversible by resveratrol supplementation.
Recommendation
Resveratrol supplementation should be incorporated as part of dietary nutrients for patients on cisplatin during chemotherapy. It is recommended, that further studies be conducted to fully ascertain the mechanisms of action of resveratrol and its anti-cisplatin properties.
Disclosures: There is no conflict of interest for all authors.
References
1. Markovic SD, Djacic DS, Cvetkovic DM, Obradovic, A.D., Zizic, J.B., Ognjanovic, BI, et al. Effects of acute in vivo cisplatin and selenium treatment on hematological and oxidative stress parameters in red blood cells of rats. Biol. Trace. Elem. Res. 2010; DOI 10.1007/s12011010-8788-9.
2. Taguchi NA, Abid MR et al. Cisplatin-associated nephrotoxicity and pathological events. Contrib Nephrol. 2005; 148:107-121.
3. Wood PA, Hrusheeky WJM. Cisplatin-associated anemia: an erythropoietin deficiency syndrome. J Clin Invest. 1995; 95:16501659.
4. Von Hoff DD, Schilsky R, Richert CM. Toxic effects of cis-dichlorodiammine platinum (II) in man. Cancer Treat. Rep. 1979; 63, 1527-1531.
5. Philipps KA, Tannock IF. Design and interpretation of clinical trials that evaluate agents that may offer protection from the toxic effects of cancer chemotherapy. J. Clin. Oncol. 1998; 16, 3179-3190.
6. Hi§miogullari SE, Hi§miogullari AA, Yavuz MT. The protective effect of resveratrol in experimentally induced non-sterile clean wound inflammation in rats. Kafkas universitesi Veteriner Fakultesi Dergisi 2013; 19, supplement A, pp. A1-A5.
7. Loehrer PJ, Gonin R, Nichols CR, Weathers T, Einhorn LH. Vinblastine plus ifosphamide plus cisplatin as initial salvage therapy in recurrent germ cell tumor. J. Clin. Oncol. 1998; 16:2500-2504.
8. Bolis G, Favalli G, Danese S, Zanaboni F, Mangili G, Scarabelli C, et al. Weekly cisplatin given for 2 months versus cisplatin plus cyclophosphamide given for 5 months after cytoreductive surgery for advanced ovarian cancer. J. Clin. Oncol. 1997;15:1938-1944.
9. Rose P, Bundy B, Watkins E, Thigpen J, Deppe G, Maiman M, et al. Concurrent cisplatin-based radiotherapy and chemotherapy for locally advanced cervical cancer. N. Engl. J. Med. 1999;340:1144-1153.
10. Coppin C, Gospodarowicz M, James K, Tannock I, Zee B, Carson J, et al. Improved local control of invasive bladder cancer by concurrent cisplatin and preoperative or definitive radiation. J. Clin. Oncol. 1996;14:2901-2907.
11. Planting A, Catimel G, de Mulder P, de Graeff A, Hoppener F, Verweij J, et al. Randomized study of a short course of weekly cisplatin with or without amifostine in advanced head and neck cancer. Ann. Oncol. 1999;10:693-700.
12. Levi JA, Aroney RS, Dalley DN. Haemolytic anaemia after cisplatin treatment. BMJ. 1981; 282: 2003- 2004.
13. Maloisel F, Kurtz JE, Andres E, Gorodetsky C, Dufour P, Oberling F. Platin salts-induced haemolytic anaemia: cisplatin and the first case of carboplatin-induced haemolysis. Anticancer drugs. 1995; 6(2): 324 - 326.
14. Donnelly LE, Newton R, Kennedy GE. Anti inflammatory effects of resveratrol in lung epithelial cells: molecular mechanisms. The American Journal of Physiology: Lung cellular and molecular physiology. 2004; 287(4): L774-L783.
15. Olas B, Wachowicz B, Majsterek I, Blasiak J. Resveratrol may reduce oxidative stress induced by platinum compounds in human plasma, blood platelets and lymphocytes. Anticancer Drugs. 2005; 16: 659-665.
16. Mukai K, Galli SJ. Basophils. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net 2013 [doi: 10.1002/9780470015902. a0001120.pub3]
17. Calvert AH, Nevell DR. Cisplatin triggers platelet activation. Journal of Clinical Oncology, 1989; 7(11):1748.
How to cite this article: Izuchukwu Azuka Okafor, Johnson Okwudili Nweke, Uchenna Somtochukwu Nnamah. Hematological dynamics following the co-administration of Resveratrol and Cisplatin in Sprague-Dawley rats. J Clin Med Kaz. 2018; 2(48):22-27
