Cardiac damage following renal ischemia reperfusion injury increased with excessive consumption of high fat diet but enhanced the cardiac resistance to reperfusion stress in rat

Fig. 1. Cardiac changes associated with SD and HD feeding in rats. Gene expression of A) ANP, B) BNP and C) Heart weight/body weight ratio *p < 0.05 vs SD. The data are presented as mean ± SD (n = 6/group). One-way ANOVA, followed by post-Dunnet's test was used to analyse the data. SD-Standard diet, HD-High fat diet.

Fig. 2. Renal Injury in SD and HD rats subjected to IR. A) Representative H and E stained images (n = 3) (The images are presented at 10x magnification) B) Renal injury score C) Urinary levels of Kim-1, D) Na–K ATPase activity, E) Plasma creatinine and F) Blood urea nitrogen. *p < 0.05 vs SD, #p < 0.05 vs HD. The data are presented as mean ± SD (n = 6/group). One-way ANOVA, followed by post-Dunnet's test was used to analyse the data. SD- Standard diet, HD-High fat diet.

Fig. 3. A) Representative H and E images of different experimental groups (n = 3) (The images are presented at 10x magnification). SD-Standard diet, HD-High fat diet, HS- Heart Sham, HR- Heart ischemia reperfusion, RS- Renal sham, RR- Renal ischemia reperfusion.

Fig. 4. A) Representative TUNNEL images of different experimental groups (n = 3) (The images are presented at 10x magnification). Biochemical analysis of B) LDH activity in the tissue C) LDH activity in the perfusate D) CK activity in the tissue and E) CK activity in the perfusate. *p < 0.05 HD group vs respective SD group, #p < 0.05 vs SD_RSHS, $ p < 0.05 vs SD_RRHS and & p < 0.05 vs HD_RRHS. The data are presented as mean ± SD (n = 6/group). One-way ANOVA, followed by post-Dunnet's test was used to analyse the data. SD-Standard diet, HD-High fat diet, HS- Heart Sham, HR- Heart ischemia reperfusion, RS- Renal sham, RR- Renal ischemia reperfusion.

Fig. 5. Spectrophotometric analysis of oxidative stress in tissue homogenate and mitochondria. A) TBARS B) GSH: GSSG ratio, C) catalase activity, D) SOD activity in the respective groups and E) DCFH/DA analysis in the tissue. *p < 0.05 HD group vs respective SD group, #p < 0.05 vs SD_RSHS, $ p < 0.05 vs SD_RRHS and & p < 0.05 vs HD_RRHS. The data are presented as mean ± SD (n = 6/group). One-way ANOVA, followed by post-Dunnet's test was used to analyse the data. SD-Standard diet, HD-High fat diet, HS- Heart Sham, HR- Heart ischemia reperfusion, RS- Renal sham, RR- Renal ischemia reperfusion.

Fig. 6. Mitochondrial electron transport chain enzyme activities in cardiac mitochondrial subpopulations using spectrophotometry-based analysis A) NQR (Complex I), B) SQR (Complex II), C) QCR (Complex III), D) COX (Complex IV) activities and E) ATP level in the mitochondrial samples isolated from heart tissues. *p < 0.05 HD group vs respective SD group, #p < 0.05 vs SD_RSHS, $ p < 0.05 vs SD_RRHS and & p < 0.05 vs HD_RRHS. The data are presented as mean ± SD (n = 6/group). One-way ANOVA, followed by post-Dunnet's test was used to analyse the data. SD-Standard diet, HD-High fat diet, HS- Heart Sham, HR- Heart ischemia reperfusion, RS- Renal sham, RR- Renal ischemia reperfusion.

Fig. 7. Gene expression analysis of A) TNF-α and B) IL-6 in the cardiac tissue, protein levels of C) TNF- α and D) IL-6 in the cardiac tissue. PI3k/Akt signaling pathway determined by qRTPCR technique. A) mRNA expression changes of PI3k B) mRNA expression changes of Akt. *p < 0.05 HD group vs respective SD group, #p < 0.05 vs SD_RSHS, $ p < 0.05 vs SD_RRHS and & p < 0.05 vs HD_RRHS. The data are presented as mean ± SD (n = 6/group). One-way ANOVA, followed by post-Dunnet's test was used to analyse the data. SD-Standard diet, HD-High fat diet, HS- Heart Sham, HR- Heart ischemia reperfusion, RS- Renal sham, RR- Renal ischemia reperfusion.

Fig. 8. Expression analysis of PI3K/AKT pathway A) Representative blot image of p-PI3k, Total PI3K, p-AKT (Ser 473), Total AKT and β actin proteins, B) Relative protein expression of p-PI3k/PI3K, C) Relative protein expression of p-AKT/AKT in the cardiac tissue *p < 0.05 HD group vs respective SD group, #p < 0.05 vs SD_RSHS, $ p < 0.05 vs SD_RRHS and & p < 0.05 vs HD_RRHS. The data are presented as mean ± SD (n = 3/group). One-way ANOVA, followed by post-Dunnet's test was used to analyse the data. SD-Standard diet, HD-High fat diet, HS- Heart Sham, HR- Heart ischemia reperfusion, RS- Renal sham, RR- Renal ischemia reperfusion. The uncropped blot images are included in the supplementary file.