{"id":59776,"date":"2024-09-30T10:30:08","date_gmt":"2024-09-30T10:30:08","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=59776"},"modified":"2024-10-09T18:42:19","modified_gmt":"2024-10-09T18:42:19","slug":"antiinflammatory-and-antioxidant-mediated-nephroprotection-of-melatonin-and-rosuvastatin-in-carboplatin-induced-nephrotoxicity-an-experimental-study","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol17no3\/antiinflammatory-and-antioxidant-mediated-nephroprotection-of-melatonin-and-rosuvastatin-in-carboplatin-induced-nephrotoxicity-an-experimental-study\/","title":{"rendered":"Antiinflammatory and Antioxidant Mediated Nephroprotection of Melatonin and Rosuvastatin in Carboplatin Induced Nephrotoxicity: An Experimental Study."},"content":{"rendered":"\n

Introduction<\/strong><\/p>\n\n\n\n

Carboplatin (cis-diammine-1,1-cyclobutanedicarboxylateplatinum II), a platinum coordination compound and alkylating agent is used in ovarian, head, neck and lung carcinomas. Carboplatin has been reported to cause hematological side effects like anemia, neutropenia leucopoenia, and minimal nephrotoxicity. 1<\/sup>The renal effects of carboplatin have been reported in high dose regimens and in the presence of concomitant risk factors.2 <\/sup>Though nephrotoxicity is dose related in both cisplatin- and carboplatin, carboplatin is less nephrotoxic but still leads to elevated serum creatinine in 10% of the patients.3 <\/sup>Cumulative doses of carboplatin have shown evidence of renal damage qualitatively like but less severe than that caused by cisplatin. Authors opine that proper monitoring of patients is required if high doses of carboplatin are used or carboplatin is combined with other nephrotoxic chemotherapy.4<\/sup> However, there is a need for drugs that can prevent carboplatin-induced nephrotoxicity. <\/p>\n\n\n\n

Melatonin is currently used in treatment of jet lag and readjustment of sleep wake cycle of shift workers. It gives protection against oxidative damage secondary to various factors like strenuous exercise, toxins primarily by scavenging the free radicals.5<\/sup> Its role in attenuating the nephrotoxicity induced by drugs like vancomycin, amikacin, and cisplatin which cause oxidative stress in kidneys has been reported.6<\/sup> Rosuvastatin, a hypolipidemic drug, decreases cholesterol levels by inhibiting the enzyme 3HMGCoA (3-hydroxy-3-methylglutaryl coenzyme) reductase. In addition, rosuvastatin has anti-inflammatory, immunomodulatory and antioxidant activities. Rosuvastatin like other statins has been reported to attenuate inflammation through inhibition of inflammatory cytokines as well as recruitment of inflammatory cells. Studies have shown the protective role of rosuvastatin in cisplatin-induced nephrotoxicity.7<\/sup><\/p>\n\n\n\n

In\nconsideration of the above reports the present experimental study was done to\nassess and compare the nephroprotective effect of melatonin and rosuvastatin at\nlow and high doses in carboplatin induced nephrotoxicity in rats.<\/p>\n\n\n\n

Material and Methods<\/strong><\/p>\n\n\n\n

Animals<\/strong><\/p>\n\n\n\n

The study was conducted after approval by the Institutional Animal Ethics Committee (IAEC\/KMC\/109\/2020). Animals were obtained from the Central Animal House of the institute in accordance with the committee for Control and supervision of experimentation on animal (CPCSEA) guidelines.Thirty-six female albino Wistar rats of weight ranging 150-250g, aged around 8-10 weeks were used for the study. The animals were housed under standard conditions, 12-hour light-dark cycle, 50% humidity, and 28\u00b0C temperature and provided with food and water ad libitum.  <\/em><\/p>\n\n\n\n

Drugs\/Chemicals<\/strong><\/p>\n\n\n\n

Carboplatin (manufactured by Fresenius Kabi\nOncology Ltd), Melatonin 3mg tab (manufactured in India by Sun Pharma\nLaboratories Ltd.), and Rosuvastatin 20mg tab (manufactured in India by Sun\nPharma Laboratories Ltd.) was purchased from Manipal Hospital Pharmacy. All the\nother reagents used were purchased from Coral Clinical Systems and were of\nanalytical grade. Dimethylsuphoxide (DMSO) was used as vehicle. The doses of\ndrugs were selected as per previous studies.5,8,9<\/sup><\/p>\n\n\n\n

Experimental procedure<\/strong><\/p>\n\n\n\n

In the experiment, a total of 36 rats were randomly divided into 6\ngroups (n=6). The different groups received treatments as mentioned in Table 1.\nAll groups other than control group were administered Carboplatin 128mg\/kg intraperitoneal\nsingle dose on day 5 to induce nephrotoxicity. <\/p>\n\n\n\n

The\nanimals were weighed on day 0 and 11. Retro orbital blood collection from inner\ncanthus of the eye using capillary tubes was done on day 0, 7 and 11 which was\nused for biochemical and antioxidant estimations. After 11th day, kidneys were\ndissected and weighed. The renal tissue was used for histopathological\nanalysis.<\/p>\n\n\n\n

Table 1: Treatment groups <\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n
\n

Group1 (normal control)<\/p>\n<\/td>\n

\n

Dimethylsulfoxide (DMSO) as vehicle from day 0 to day 10 once a day<\/p>\n<\/td>\n<\/tr>\n

\n

Group II (Carboplatin control)<\/p>\n<\/td>\n

\n

DMSO: 1ml\/kg\/day p.o for 10 days from day 0 to day 10 once a day<\/p>\n<\/td>\n<\/tr>\n

\n

Group III (Carboplatin + Melatonin low dose)<\/p>\n

(Car +L.M)<\/p>\n<\/td>\n

\n

Melatonin: 5mg\/kg p.o for 10 days from day 0 to day 10 once a day<\/p>\n<\/td>\n<\/tr>\n

\n

Group IV (Carboplatin + Melatonin high dose)<\/p>\n

(Car +H.M)<\/p>\n<\/td>\n

\n

Melatonin: 10mg\/kg p.o for 10 days from day 0 to day 10 once a day<\/p>\n<\/td>\n<\/tr>\n

\n

Group V (Carboplatin + Rosuvastatin low dose)<\/p>\n

(Car +L.R)<\/p>\n<\/td>\n

\n

Rosuvastatin: 10mg\/kg p.o for 10 days from day 0 to day 10 once a day <\/p>\n<\/td>\n<\/tr>\n

\n

Group VI (Carboplatin + Rosuvastatin high dose)<\/p>\n

(Car +H.R)<\/p>\n<\/td>\n

\n

Rosuvastatin: 20mg\/kg p.o for 10 days from day 0 to day 10 once a day <\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

Biochemical estimations<\/strong><\/p>\n\n\n\n

Blood Urea was estimated usingGLDH kinetic method10<\/sup> and serum creatinine was estimated by modified\nJaffe\u2019s Kinetic method.11<\/sup>Uric\nacid estimation was done by uricase\/PAP method.12 <\/sup>Serum IL-18 was measured using ELISA.13<\/sup>\nThe antioxidants glutathione (GSH) and malondialdehyde (MDA) were assayed using\nmethods described earlier.9<\/sup><\/p>\n\n\n\n

Histopathological Examination<\/strong><\/p>\n\n\n\n

The histological examination of kidney tissues\nwas done using paraffin-embedded specimens. The specimens were cut into 6 mm thickness and stained with\nHematoxylin-Eosin for light microscope examination (Olympus, BH-2, Tokyo,\nJapan). The characteristic histological changes in sections was observed and\nwere analyzed semi-quantitatively using the technique of Houghton et al. 14<\/sup>\nThe lesions were graded as 0,1 or 2 if the histology showed normal structure,\nareas of focal granular-vacuolar epithelial cell degeneration or tubular\nepithelial necrosis respectively<\/p>\n\n\n\n

Results <\/strong><\/p>\n\n\n\n

The effect of melatonin and Rosuvastatin on body\nweight and weight of kidneys in carboplatin induced nephrotoxicity is shown in\ntable 2. No significant difference was observed in\nthe weight of animals on day 0 and day 11. There was no significant change in\nthe weight of kidneys in treatment groups as compared to control.<\/p>\n\n\n\n

Biochemical estimations<\/strong><\/p>\n\n\n\n

Blood urea<\/strong><\/p>\n\n\n\n

The blood urea in six groups is shown in Table 3. On day 11 Carboplatin showed a significant increase in blood urea levels as compared to control (p<0.05). Also on day 11, all treatment groups showed a significant decrease in blood urea levels as compared to the carboplatin group (Table 3)               <\/p>\n\n\n\n

Serum creatinine<\/strong><\/p>\n\n\n\n

 The serum creatinine levels of different groups is shown in table 4. On day 11 carboplatin group showed a significant increase in serum creatinine levels as compared to control(p<0.05). There is a significant decrease in creatinine levels as compared to carboplatin in all treatment groups. On comparing day 11 and day 7 levels with day 0 in carboplatin group, a significant increase is observed on day 11 and day 7 (p<0.05).    <\/strong><\/p>\n\n\n\n

Uric acid<\/strong><\/p>\n\n\n\n

On day 11 carboplatin group showed a significant increase in uric acid levels as compared to control(p<0.05). There was a significant decrease in uric acid levels as compared to carboplatin in rosuvastatin and high dose of melatonin treatment groups (table 5). On comparing day 11 and day 7 levels with day 0 in carboplatin group, a significant increase is observed on day 11 and day 7 (p<0.05). <\/strong><\/p>\n\n\n\n

IL-18 levels<\/strong><\/p>\n\n\n\n

The IL-18 levels were significantly more in carboplatin group on day11 as compared to control group (p<0.05). On day 11, all treatment groups showed a decrease in IL-18 levels as compared to carboplatin but the decrease was not statistically significant (p>0.05) as seen in table 6. <\/p>\n\n\n\n

Antioxidant estimation<\/strong><\/p>\n\n\n\n

GSH assay<\/strong><\/p>\n\n\n\n

There is a non- significant decrease in GSH levels in carboplatin treated group as compared to control on day11 (table 7). GSH levels in the other treatment groups were comparable to control (p>0.05) <\/p>\n\n\n\n

MDA assay<\/strong><\/p>\n\n\n\n

There\nwas significant increase in MDA levels in carboplatin treated group in\ncomparison to the control group. The MDA levels in the treatment groups were\nsignificantly reduced in comparison to the carboplatin treated group as shown\nin table 8. <\/p>\n\n\n\n

Table 2: Body weight of rats on day 0 and day 11 and weight of kidney on day 11<\/strong>.<\/p>\n\n\n\n\n\n\n\n\n\n\n\n
\n

GROUPS<\/strong><\/p>\n<\/td>\n

\n

Body weight in (g) DAY0<\/strong><\/p>\n<\/td>\n

\n

Body weight in (g) DAY11<\/strong><\/p>\n<\/td>\n

\n

Kidney weight (g)<\/strong><\/p>\n

Day 11<\/strong><\/p>\n<\/td>\n

\n

Organ weight\/Bodyweight<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

1.Control<\/p>\n<\/td>\n

\n

153.33\u00b18.16<\/p>\n<\/td>\n

\n

182.00\u00b116.92<\/p>\n<\/td>\n

\n

0.62 \u00b1 0.074<\/p>\n<\/td>\n

\n

0.003<\/p>\n<\/td>\n<\/tr>\n

\n

2.Carboplatin<\/p>\n<\/td>\n

\n

175.00\u00b113.52<\/p>\n<\/td>\n

\n

172.67\u00b110.07<\/p>\n<\/td>\n

\n

0.59 \u00b1 0.028<\/p>\n<\/td>\n

\n

0.003<\/p>\n<\/td>\n<\/tr>\n

\n

3.Car+ L.M<\/p>\n<\/td>\n

\n

174.00\u00b125.27<\/p>\n<\/td>\n

\n

164.00\u00b126.58<\/p>\n<\/td>\n

\n

0.67 \u00b1 0.079<\/p>\n<\/td>\n

\n

0.003<\/p>\n<\/td>\n<\/tr>\n

\n

4.Car +H.M<\/p>\n<\/td>\n

\n

177.33\u00b120.20<\/p>\n<\/td>\n

\n

182.67\u00b132.40<\/p>\n<\/td>\n

\n

0.65 \u00b1 0 .096<\/p>\n<\/td>\n

\n

0.003<\/p>\n<\/td>\n<\/tr>\n

\n

5.Car +L.R<\/p>\n<\/td>\n

\n

184.17\u00b119.33<\/p>\n<\/td>\n

\n

182.67\u00b132.40<\/p>\n<\/td>\n

\n

0.70 \u00b1 0.083<\/p>\n<\/td>\n

\n

0.003<\/p>\n<\/td>\n<\/tr>\n

\n

6.Car +H.R<\/p>\n<\/td>\n

\n

187.33\u00b124.54<\/p>\n<\/td>\n

\n

173.33\u00b128.92<\/p>\n<\/td>\n

\n

0.64 \u00b1 0.062<\/p>\n<\/td>\n

\n

0.003<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

            <\/em>All values are expressed as Mean\u00b1S.D<\/p>\n\n\n

Table 3: Blood urea levels in different groups on day 0, 7 and 11.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n
\n

GROUPS<\/strong><\/p>\n<\/td>\n

\n

Urea at DAY 0<\/strong><\/p>\n

mg\/dl<\/strong><\/p>\n<\/td>\n

\n

Urea at DAY7<\/strong><\/p>\n

mg\/dl<\/strong><\/p>\n<\/td>\n

\n

Urea at DAY11<\/strong><\/p>\n

mg\/dl<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

1.Control<\/p>\n<\/td>\n

\n

1.06 \u00b1 0.07<\/p>\n<\/td>\n

\n

1.35 \u00b1 0 .33<\/p>\n<\/td>\n

\n

1.21\u00b1 0.13a<\/sup><\/p>\n<\/td>\n<\/tr>\n

\n

2.Carboplatin<\/p>\n<\/td>\n

\n

1.23 \u00b1 0.13<\/p>\n<\/td>\n

\n

3.07 \u00b10.28a<\/sup><\/p>\n<\/td>\n

\n

3.38\u00b1 0.13*a<\/sup><\/p>\n<\/td>\n<\/tr>\n

\n

3.Car+ L.M<\/p>\n<\/td>\n

\n

1.17  \u00b1 0.15<\/p>\n<\/td>\n

\n

1.56 \u00b10 .36a<\/sup><\/p>\n<\/td>\n

\n

1.22\u00b1 0.11+<\/sup><\/p>\n<\/td>\n<\/tr>\n

\n

4.Car +H.M<\/p>\n<\/td>\n

\n

1.01  \u00b1 0.27<\/p>\n<\/td>\n

\n

1.46 \u00b1 0. 21a<\/sup><\/p>\n<\/td>\n

\n

1.15 \u00b1 0.19+<\/sup><\/p>\n<\/td>\n<\/tr>\n

\n

5.Car +L.R<\/p>\n<\/td>\n

\n

1.23  \u00b1 0.03<\/p>\n<\/td>\n

\n

1.73 \u00b1 0.54<\/p>\n<\/td>\n

\n

1.15 \u00b1 0.20+<\/sup><\/p>\n<\/td>\n<\/tr>\n

\n

6.Car +H.R<\/p>\n<\/td>\n

\n

1.24  \u00b1 0.03<\/p>\n<\/td>\n

\n

1.25 \u00b10 .35<\/p>\n<\/td>\n

\n

1.06 \u00b1 0.32+<\/sup><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Repeated measures of ANOVA p<0.05, post hoc Bonferronis test *<\/sup>p<0.05 vs control, a <\/sup>vs day 0, One way ANOVA on day 11, post hoc Tukey test +<\/sup>p<0.05 vs carboplatin. All values are expressed   as Mean\u00b1S.D<\/p>\n\n\n

Table 4: Serum creatinine levels in different groups on day0, 7 and 11.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n
\n

GROUPS<\/strong><\/p>\n<\/td>\n

\n

Creatinine at DAY 0<\/strong><\/p>\n

mg\/dl<\/strong><\/p>\n<\/td>\n

\n

 Creatinine at DAY7<\/strong><\/p>\n

 mg\/dl<\/strong><\/p>\n<\/td>\n

\n

Creatinine at DAY11 <\/strong><\/p>\n

mg\/dl<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

1.Control<\/p>\n<\/td>\n

\n

0.24 \u00b1 0.03<\/p>\n<\/td>\n

\n

0.23 \u00b1 0.03<\/p>\n<\/td>\n

\n

0.24 \u00b1 0.03<\/p>\n<\/td>\n<\/tr>\n

\n

2.Carboplatin<\/p>\n<\/td>\n

\n

0.24 \u00b1 0.03<\/p>\n<\/td>\n

\n

1.60 \u00b10 .66a<\/sup><\/strong><\/p>\n<\/td>\n

\n

1.86 \u00b1 0.24*a<\/sup><\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

3.Car+ L.M<\/p>\n<\/td>\n

\n

0.23 \u00b1 0.03<\/p>\n<\/td>\n

\n

0.64 \u00b10 .40<\/p>\n<\/td>\n

\n

0.28 \u00b1 0.09+<\/sup><\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

4.Car +H.M<\/p>\n<\/td>\n

\n

0.24 \u00b10. 03<\/p>\n<\/td>\n

\n

0.37 \u00b1 0.21<\/p>\n<\/td>\n

\n

0.22 \u00b1 0.11+<\/sup><\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

5.Car +L.R<\/p>\n<\/td>\n

\n

0.23 \u00b1 0.03<\/p>\n<\/td>\n

\n

0.38 \u00b1 0.20<\/p>\n<\/td>\n

\n

0.26 \u00b1 0 .12+<\/sup><\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

6.Car +H.R<\/p>\n<\/td>\n

\n

0.24 \u00b1 0.03<\/p>\n<\/td>\n

\n

0.32 \u00b1 0.27<\/p>\n<\/td>\n

\n

0.22 \u00b1 0.14+<\/sup><\/strong><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Repeated measures of ANOVA, post hoc Bonferroni\u2019s test p<0.05, *vs control, + <\/sup>vs carboplatin, One way ANOVA on day 11, post hoc Tukey test , a <\/sup>vs day 0. All values are expressed as Mean\u00b1S.D<\/p>\n\n\n

Table 5: Uric acid levels in different groups on day 0, 7 and 11.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n
\n

GROUPS<\/strong><\/p>\n<\/td>\n

\n

Uric acid at DAY 0 <\/strong><\/p>\n

mg\/dl<\/strong><\/p>\n<\/td>\n

\n

Uric acid at DAY7<\/strong><\/p>\n

mg\/dl<\/strong><\/p>\n<\/td>\n

\n

Uric acid at DAY11<\/strong><\/p>\n

mg\/dl<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

1.Control<\/p>\n<\/td>\n

\n

0.70 \u00b1 0.21<\/p>\n<\/td>\n

\n

0.82 \u00b10.34<\/p>\n<\/td>\n

\n

0.96 \u00b10 .18<\/p>\n<\/td>\n<\/tr>\n

\n

2.Carboplatin<\/p>\n<\/td>\n

\n

0.48 \u00b10.17<\/p>\n<\/td>\n

\n

1.40 \u00b10.21a<\/sup><\/strong><\/p>\n<\/td>\n

\n

2.57 \u00b1 1.07*a<\/sup><\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

3.Car+ L.M<\/p>\n<\/td>\n

\n

0.82 \u00b1 0.43<\/p>\n<\/td>\n

\n

1.37 \u00b10.64<\/p>\n<\/td>\n

\n

1.93 \u00b1 0.54<\/p>\n<\/td>\n<\/tr>\n

\n

4.Car +H.M<\/p>\n<\/td>\n

\n

0.89 \u00b1 0.29<\/p>\n<\/td>\n

\n

1.01 \u00b10.40<\/p>\n<\/td>\n

\n

0.88 \u00b10 .38+<\/sup><\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

5.Car +L.R<\/p>\n<\/td>\n

\n

0.63 \u00b1 0.23<\/p>\n<\/td>\n

\n

0.82 \u00b10.43<\/p>\n<\/td>\n

\n

1.01 \u00b1 0.40+<\/sup><\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

6.Car +H.R<\/p>\n<\/td>\n

\n

0.75 \u00b1 0.38<\/p>\n<\/td>\n

\n

0.76 \u00b10.29<\/p>\n<\/td>\n

\n

0.88 \u00b1 0.45+<\/sup><\/strong><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Repeated measure of ANOVA, post hoc Bonferroni\u2019s test *p<0.05 vs day 0, + <\/sup>vs carboplatin. One way ANOVA on day 11, post hoc Tukey test a<\/sup>p<0.05 vs day 0. All values are expressed as Mean\u00b1S.D<\/p>\n\n\n

Table 6: IL-18 levels on day0, 7 and 11.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n
\n

GROUPS<\/strong><\/p>\n<\/td>\n

\n

DAY0<\/strong><\/p>\n

ng\/l<\/strong><\/p>\n<\/td>\n

\n

DAY7<\/strong><\/p>\n

ng\/l<\/strong><\/p>\n<\/td>\n

\n

DAY11<\/strong><\/p>\n

ng\/l<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

1.Control<\/p>\n<\/td>\n

\n

11.83 \u00b1 0.72<\/p>\n<\/td>\n

\n

11.91 \u00b1 0.79<\/p>\n<\/td>\n

\n

11.89 \u00b1 0.76<\/p>\n<\/td>\n<\/tr>\n

\n

2.Carboplatin<\/p>\n<\/td>\n

\n

11.11 \u00b1 0.84<\/p>\n<\/td>\n

\n

14.06 \u00b1 0.82<\/p>\n<\/td>\n

\n

14.63 \u00b1 0.82*<\/sup><\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

3.Car+ L.M<\/p>\n<\/td>\n

\n

11.24 \u00b1 0.76<\/p>\n<\/td>\n

\n

12.95 \u00b1 0.52<\/p>\n<\/td>\n

\n

13.49 \u00b1 0.94<\/p>\n<\/td>\n<\/tr>\n

\n

4.Car +H.M<\/p>\n<\/td>\n

\n

10.91 \u00b1 0.64<\/p>\n<\/td>\n

\n

13.38 \u00b1 0.75<\/p>\n<\/td>\n

\n

12.78 \u00b1 1.30<\/p>\n<\/td>\n<\/tr>\n

\n

5.Car +L.R<\/p>\n<\/td>\n

\n

11.77 \u00b1 0.53<\/p>\n<\/td>\n

\n

12.84 \u00b1 0.48<\/p>\n<\/td>\n

\n

13.38 \u00b1 1.11<\/p>\n<\/td>\n<\/tr>\n

\n

6.Car +H.R<\/p>\n<\/td>\n

\n

11.76 \u00b1 0.68<\/p>\n<\/td>\n

\n

12.19 \u00b1 0.94<\/p>\n<\/td>\n

\n

12.86 \u00b1 1.55<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Repeated measures of ANOVA, post hoc Bonferroni\u2019s test *p<0.05 <\/sup><\/strong>vscarboplatin. Remaining groups were tested but not significantly comparable to control group. All values are expressed as Mean\u00b1S.D.<\/p>\n\n\n

Table 7: GSH levels in kidney tissue of rats<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n
\n

GROUPS<\/strong><\/p>\n<\/td>\n

\n

GSH levels in \u00b5M\/mg tissue<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

1.Control<\/p>\n<\/td>\n

\n

1.312 \u00b1 0.13<\/p>\n<\/td>\n<\/tr>\n

\n

2.Carboplatin<\/p>\n<\/td>\n

\n

1.235 \u00b1 0.13<\/p>\n<\/td>\n<\/tr>\n

\n

3.Car+ L.M<\/p>\n<\/td>\n

\n

1.463 \u00b1 0.26<\/p>\n<\/td>\n<\/tr>\n

\n

4.Car +H.M<\/p>\n<\/td>\n

\n

1.538 \u00b1 0.46<\/p>\n<\/td>\n<\/tr>\n

\n

5.Car +L.R<\/p>\n<\/td>\n

\n

1.354 \u00b1 0.35<\/p>\n<\/td>\n<\/tr>\n

\n

6.Car +H.R<\/p>\n<\/td>\n

\n

1.248 \u00b1 0.19<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

  All values are expressed as Mean\u00b1S.D<\/p>\n\n\n

Table 8: MDA levels in kidney tissue of rats.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n
\n

GROUPS<\/strong><\/p>\n<\/td>\n

\n

MDA levels in nM\/mg tissue<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

1.Control<\/p>\n<\/td>\n

\n

51.15 \u00b1 0.599<\/p>\n<\/td>\n<\/tr>\n

\n

2.Carboplatin<\/p>\n<\/td>\n

\n

63.83 \u00b1 1.23*<\/sup><\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

3.Car+ L.M<\/p>\n<\/td>\n

\n

54.73\u00b13.31+<\/sup><\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

4.Car +H.M<\/p>\n<\/td>\n

\n

48.61\u00b15.89+<\/sup><\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

5.Car +L.R<\/p>\n<\/td>\n

47.32\u00b14.61+<\/sup><\/strong><\/td>\n<\/tr>\n
\n

6.Car +H.R<\/p>\n<\/td>\n

\n

47.33\u00b15.19+<\/sup><\/strong><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

<\/p>\n\n\n<\/p>\n

All values are expressed as Mean\u00b1S.D<\/p>\n

\n\n\n\n<\/p>\n

One way ANOVA, p<0.05; post hoc Tukey test, *<\/strong>p<0.05 vs control, +<\/sup><\/strong>p<0.05 vs carboplatin<\/span>    <\/strong>\n\n\n

<\/p>\n\n\n

Histopathological examinations<\/strong><\/p>\n\n\n\n

Microphotographs of rat kidneys stained by Hematoxylin and Eosin (200X) is shown in the figure 1.  <\/p>\n\n\n\n

  In control group (A) there are no changes in\nglomerulus, tubules and vessels, but there was mild small lymphocytic\ninfiltration in interstitium. In carboplatin group (B) glomerular atrophy,\nvacuolar degeneration, hyaline casts were observed in the tubules. In car+ L.M\ngroup (C) glomerular atrophy, vacuolar degeneration, hyaline casts in   tubules and moderate to severe small\nlymphocytic infiltration was seen in interstitium, but no changes were observed\nin the vessels.  In car+ H.M (D)\ninfiltration by small lymphoid cells are present. In car+ L.R (E) RBC casts\nseen inside the tubules. In Car + H.R (F) RBC casts seen inside the tubules\nalong with glomerular atrophy and vacuolar degeneration but had no changes in\ninterstitium and vessels. <\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 1<\/strong>: Histopathology of renal tissue in different treatment groups<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Discussion<\/strong><\/p>\n\n\n\n

In\nthe present study, nephroprotective effects of melatonin and rosuvastatin in\ncarboplatin induced nephrotoxic model were evaluated. Carboplatin 128mg\/kg\nsingle dose i.p induced nephrotoxicity which was manifested as increase in\nblood urea, serum creatinine and serum uric acid levels. This was further\nconfirmed by the glomerular atrophy, vacuolar degeneration and hyaline casts in\nkidney tubules seen in histopathology which was similar to a previous study. 15<\/sup>\nCarboplatin caused dose-dependent renal injury in rats by depleting renal\nantioxidants, enhancing lipid peroxidation in addition to platinum content.16<\/sup>\n<\/p>\n\n\n\n

Treatment\nwith low and high doses of melatonin and carboplatin prevented the rise in\nblood urea, serum creatinine and uric acid levels on day 7 and day 11, which\nwas suggestive of their nephroprotective property. This was confirmed by\nhistopathological studies where there was only mild infiltration in\ninterstitium and no changes in blood vessels. <\/p>\n\n\n\n

Melatonin\nattenuates the nephrotoxicity induced by drugs by its antioxidant action as it\nstrengthens the antioxidant enzymes and causes free radical scavenging\ndirectly. In addition, it modulates inflammatory cytokines which in turn\nreestablishes the balance of apoptosis and cell survival.17<\/sup>  <\/p>\n\n\n\n

Treatment\nwith low and high doses of rosuvastatin and carboplatin prevented the rise in\nblood urea, serum creatinine and uric acid on day 7 and day 11 which is\nsuggestive of their nephroprotective effect as well. Histopathology of\nrosuvastatin groups did not show any changes in interstitium and vessels. This\neffect was predominantly seen at the high dose of rosuvastatin. Statins in\ngeneral produce nephroprotective effect by inhibiting receptor-mediated\nendocytosis that is responsible for the protein uptake in proximal tubular\ncells in experimental studies.18<\/sup><\/p>\n\n\n\n

Clinical\nstudies have shown that statins produce nephroprotective effect secondary to pleiotropic\neffects like antithrombotic, anti-inflammatory and decreasing endothelial\nnitric oxide synthase.19<\/sup> <\/p>\n\n\n\n

Though\nsome studies report the superiority of lipophilic statins like atorvastatin\nover rosuvastatin in terms of nephroprotective effect20<\/sup>, other\nstudies refute it and claim rosuvastatin to be better.<\/p>\n\n\n\n

Comparison\nof melatonin and rosuvastatin did not show any significant difference in terms\nof nephroprotective activity and histopathological changes. This implicates\nthat the nephroprotective action of melatonin is comparable to that of\nrosuvastatin. There was no significant difference in the nephroprotective\nactivity of different doses of rosuvastatin and melatonin except that the low\ndose of melatonin did not show a significant decrease.<\/p>\n\n\n\n

Studies\nhave shown that different statins like atorvastatin and rosuvastatin have same\nefficacy in preventing contrast induced nephropathy.21<\/sup><\/p>\n\n\n\n

IL-18\nis an inflammatory marker produced during acute kidney injury. IL-18,\nmacrophage-derived cytokine, is expresses in cells like macrophages, dendritic\ncells, T cells, and B cells in addition to parenchymal kidney cells i.e.tubular\nepithelial cells, podocytes, and mesangial cells. 22<\/sup> IL-18 plays an\nimportant role in processes that aggravate renal injury in extension phase of\nacute kidney injury. 23 <\/sup>Animal studies have shown that drugs that\ntarget IL-18\u2013signaling axis can attenuate renal injury.24<\/sup> In the\npresent study, melatonin and rosuvastatin reduced the 1L-18 levels suggesting\ntheir anti- inflammatory action. The exact mechanism of IL-18 in renal injury\nis not clear but studies have shown that it has an important role in tubular\ninjury and renal dysfunction and thus IL-18 as a target has a wide therapeutic\npotential.24<\/sup><\/p>\n\n\n\n

Melatonin\nhas shown to ameliorate antibiotic induced nephrotoxicity by reducing oxidative\nstress in renal tissues. 25<\/sup>Melatonin and its metabolites scavenge\nfree radicals. In addition to stimulating the synthesis of antioxidant enzymes\nit protects oxidative damage of antioxidant enzymes.26<\/sup> Furthermore,\nmelatonin also exerts an anti-apoptotic effect on various types of cells.27<\/sup>\nThe mechanism postulated by Dai W et al is that melatonin mediated cell\napoptosis by inhibiting the Parkin1 signaling pathway induced by NLRP3\ninflammasomes reduces the secretion of inflammatory cytokines like IL-18.28<\/sup><\/p>\n\n\n\n

Rosuvastatin\nproduced significant renoprotective effect in gentamicin induced nephrotoxicity\nin Sprague Dolly rats by reducing kidney injury molecule-1 and interleukin-18.29<\/sup>\nIn addition to antioxidant action, the renoprotective effect of rosuvastatin\ncan be mediated by lipid lowering action. prevention of lipid accumulation in\nrenal tubules can also be contributing to renoprotective action of statins.7<\/sup>\nStudies have shown that rosuvastatin produces nephroprotection by hemin-like effect on hemeoxygenase 1\nwhich has a vital role to play in oxidative stress against antioxidants. 30\n<\/sup>Also, when comparing between different statins, rosuvastatin\nshowed a significant renoprotective effect as compared to simvastatin.7<\/sup>\n<\/p>\n\n\n\n

As\noxidative stress is a major mechanism implicated in platinum compounds induced\nnephrotoxicity, malondialdehyde and reduced glutathione assay was done in\nkidney tissue to measure lipid peroxidation and glutathione content.\nCarboplatin induced a significant increase in MDA and a decrease in GSH (though\nnot significant).  A significant decrease\nin MDA by both melatonin and rosuvastatin suggests their antioxidant mechanism.31<\/sup>\nGSH is also a biomarker for oxidative stress and its increase in GSH in\ntreatment groups was comparable to control group.<\/p>\n\n\n\n

Both\nmelatonin and rosuvastatin increased GSH levels most probably by inhibiting\nlipid peroxidation<\/p>\n\n\n\n

This\nstudy has certain limitations. Urinary markers of kidney injury and other serum\nmarkers like beta 2 microglobulin which is an early marker of renal injury were\nnot estimated.<\/p>\n\n\n\n

Conclusion<\/strong><\/p>\n\n\n\n

To\nconclude, rosuvastatin and melatonin demonstrated an equi-efficacious\nnephroprotective effects mediated by anti-inflammatory and antioxidant\nproperties. The anti-inflammatory action was primarily mediated by reducing\nIL-18 and antioxidant action by decreasing MDA and increasing GSH levels. Further\nlong-term studies are warranted to substantiate these findings. <\/p>\n\n\n\n

Acknowledgement<\/strong><\/p>\n\n\n\n

The\nauthors acknowledge Kasturba Medical College, Manipal for the support and\nproviding facilities for the conduct of research.<\/p>\n\n\n\n

Conflict of Interest<\/strong><\/p>\n\n\n\n

<\/strong>The authors declare that there are no competing interests.<\/p>\n\n\n\n

Funding Source<\/strong><\/p>\n\n\n\n

The study did not receive any external funding.<\/p>\n\n\n\n

Ethics committee<\/strong><\/p>\n\n\n\n

The study was conducted after approval by the Institutional Animal Ethics Committee (IAEC\/KMC\/109\/2020). <\/p>\n\n\n\n

References<\/strong><\/p>\n\n\n\n

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    CrossRef<\/a><\/li><\/ol>\n","protected":false},"excerpt":{"rendered":"

    Introduction Carboplatin (cis-diammine-1,1-cyclobutanedicarboxylateplatinum II), a platinum coordination compound and alkylating […]<\/p>\n","protected":false},"author":15,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[117],"tags":[],"class_list":["post-59776","post","type-post","status-publish","format-standard","hentry","category-vol17no3"],"_links":{"self":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/59776","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/users\/15"}],"replies":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/comments?post=59776"}],"version-history":[{"count":5,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/59776\/revisions"}],"predecessor-version":[{"id":61734,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/59776\/revisions\/61734"}],"wp:attachment":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/media?parent=59776"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/categories?post=59776"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/tags?post=59776"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}