@2024 Afarand., IRAN

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ISSN: 2252-0805    The Horizon of Medical Sciences 2016;22(4):337-344

Background
Nephropathy is the relative decline in renal function which is associated with Nephrotic syndrome, glomerular necrosis, albuminuria, Glomerular Filtration Reduction (GFR), blood pressure increase and fluid retention [1].
Previous Studies
… [2-13]. Due to a variety of biological activities including antioxidant activity, anti-cancer effects, anti-inflammatory effects and the treatment of hyperlipidemia, turmeric is currently on attention [14].
Aim(s)
The aim of this study was to determine the effects of hydro-alcoholic extract of turmeric on Adriamycin-induced renal injury in rats.
Research type
This study is experimental.
Research society, place and time
This study was conducted in 2015 in Mashhad University of Medical Sciences.
Sampling method and number
32 male Wistar rats with the weight of 220-300 gr were used. During the experiment, the rats were kept under the standard condition, at a temperature of 22 ° ± 2 C and 22-hour cycle of light and darkness in the animal room of Medical school, and all the animals had free access to food and water.
Used devices&materials
Hydro-alcoholic extract of turmeric was prepared in the form of soaked. First, turmeric was prepared from prestigious herbal medicine store in Mashhad and it was by Herbarium of the Ferdowsi University of Mashhad. 100 gr of powder was weighted and then incubated in 900 ml of 70% Ethyl alcohol solution at the temperature of 35°C for 72 hours. After 72 hours, the resulting solution, was filtered using filter papers with different pore sizes. To remove the solvent, the resulting solution was placed in the oven at the temperature of 40 °C so that the solvent is evaporated and the dried extract is achieved [12]. In this method, the efficiency of the extract was 30%. To produce different amounts for use in the experiment, the obtained extract was dissolved in distilled water. Rats were divided randomly into four groups of eight healthy controls, Adriamycin, turmeric extract, and turmeric extract with Adriamycin; in the healthy control group, the animals on the seventh day of study, received intravenous Saline injection equal to required volume for Adriamycin injection. For Adriamycin group, single-dose administration of 5 mg per kg of Adriamycin [15] into the tail vein was conducted on the seventh day. Turmeric extract with Adriamycin group, received turmeric hydro-alcoholic extract in amount of 1000 mg per kg for 6 days prior to the injection of Adriamycin (as a single dose of 5 mg per kg) on the seventh day and then daily for 28 days orally. The sole turmeric extract group, also, received turmeric hydro-alcoholic extract in the amount of 1000 mg per kg [16] without injection of Adriamycin. The groups were treated for 5 weeks and on days zero, 6, 10, 14, 21, 28 and 35, their urine samples was collected for measuring the Glomerular Filtration Rate (GFR). Achieved sample size was measured by mesor and then urine was kept at the temperature of 20 ° C for testing in micro-tube. Urine creatinine concentration was measured with the method of JAFFE based on the relevant kit instruction (Parsazmoon, Iran) and by the photometer Model 100 (Convergys, Germany) at a wavelength of 505 nm. In this method, creatinine makes a colorful complex with alkaline Pikerate that the intensity of produced color is proportional to the amount of creatinine in the sample [17]. In order to examine urine samples, urine was diluted with the ratio of 50+1 with distilled water and ultimately resulting number was multiplied by 51. Glomerular filtration rate (creatinine clearance based on ml per minute) was calculated through “the amount of urine creatinine (mg per deciliter)x urine output (ml per minute)’ divided by “ the level of plasma creatinine (mg per deciliter)”. To determine the percentage of kidney damage, left kidneys of animals were used. After weighing the animals, their left kidney was left outside of the body and it was halved in a longitudinal incision. Then, the two halves was fixed with 10% formalin and it was placed in paraffin after the process and the slices of 3-4 micrometers were prepared. Slices were placed on a glass side and for analyzing by light microscopy, the staining was performed with hematoxylin and eosin. Renal pathological changes was recorded by a rating scale of zero to four by Goering et al. [18] in this form: zero= indistinguishable from control group, 1=minimum (less than or equal to 25% of cortex is being affected), 2=mild (greater than 25% and less or equal to 50% of cortex is being affected), 3=average (more than 50% and less or equal to 75% of cortex is being affected), and 4=severe (more than 75% of cortex is being affected). Renal pathology was investigated with regard to the percentage of renal damage according to vacuolization, inflammation, hyaline casts tubules bleeding, cell swelling, loss of tubular and glomerular integrity, glomerular space increase, glomerular atrophy, and tubular distension with a magnification of 40 (scale sample=20 micrometers) and in any slide, 3 fields were examined. Data were analyzed by SPSS 16 software. In order to compare variables between groups, ANOVA and Tukey post hoc test and to compare the variables in the groups, paired T-test were used.
Findings by Text
The mean GFR in the control group in each of days 10, 14, 21, 28, 35 was not significantly different from day zero and 6. The mean GFR in the Adriamycin group was significantly decreased only on days 21 and 35 compared to the days zero and 6. However, no significant difference was observed in the remained days. The mean GFR in the group of turmeric extract was significantly increased in each of days 10, 14, 21, and 28 compared to days zero and 6. However, no significant change was observed on day 35 compared to days zero and 6. In the group of Adriamycin with turmeric extract no significant difference was observed between the mean GFR in each days of 10, 14, 21, 28, and 35 and the days zero and 6 (table 1). Changes in body weight was significantly decreased during the study in the group receiving Adriamycin as well as the group treated with Adriamycin plus turmeric extract compared to the control group (Figure 1). Percentage of renal damage in each of Adriamycin group (p<0.001) and Adriamycin plus turmeric extract (p<0.01) was significantly increased compared to the control group. Also, the percentage of kidney damage in turmeric extract and turmeric extract plus Adriamycin showed a significant decrease compared to Adriamycin group (p<0.001; Figure 2). In examining the microscopic section of cortex area, in the group of Adriamycin plus turmeric extract relative improvement was observed in vacuolization, inflammation and vascular congestion and dilation of tubular space (Figure 1 A, 1B, 1 C and 1D).
Main comparison to the similar studies
… [17-21]. Adriamycin administration caused glomerular sclerosis and the damage and uniformity of Podocytes [22]. In another study, also, Adriamycin was used intravenously. Fragmentation and reduction in the number of the waste podocytes feet were caused by a decrease in the expression of podocytic nephron protein [23]. … [24-34].
Suggestions
It is recommended that other doses of turmeric extract with other solvents and other administration methods, also, be investigated.
Limitations
Of the limitation of this study, was time-consuming nature of preparing the extract and pathology work.
Conclusions
Hydro-alcoholic extract of turmeric reduces the percentage of Adriamycin-induced renal injury in rats and improves the GFR amount.
Acknowledgments
The authors appreciate the extensive support of the Research Deputy of University of Medical Sciences.
Conflict of interest
Non-declared
Ethical Permissions
Ethical approval of this study has been obtained from medical ethical committee of Mashhad University of Medical Sciences.
Funding Sources
This article is the result of research project and enjoyed the financial support of respectful deputy of Mashhad University of Medical Sciences.

TABLES and CHARTS

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