Immunohistochemical analysis of TLR2 expression level in rat renal tissues in different age groups.

<p>Immunohistochemical analysis of TLR2 expression level in rat renal tissues in different age groups.</p

Changes in the kidney function and metabolic function in the aging rats.

<p>Compared with the 3-month-old group;</p><p>* indicates P<0.05; compared with the 12-month-old group;</p>#<p>indicates P<0.05.</p

Activation changes of the MyD88-independent downstream signaling pathway molecule IRF-3 in the aging kidneys.

<p>(<b>A</b>) Western blotting was used to detect the expression levels of the Phospho-IRF-3 protein in various groups of rats. (<b>B</b>) Quantitative gray scale analysis results. Compared with the 3-month-old group, * indicates P<0.05; compared with the 12-month-old group, # indicates P<0.05.</p

Expression of endogenous ligands of TLRs in rat kidney tissues in different age groups.

<p>(<b>A</b>) Western blot detection of the HSP70 and HMGB1 protein expression levels in various groups of rats. (<b>B</b>) Quantitative analysis of gray scales for the HMGB1 and HSP70 expression levels. β-actin: internal reference. Compared with the 3-month-old group, * indicates P<0.05.</p

RT-qPCR primer sequences and the lengths of the products.

<p>F, forward; R, reverse; IL-6: interleukin-6; CCL3: chemokine ligand 3; CCL4: chemokine ligand 4; CCL5: chemokine ligand 5; ICAM-1: intercellular adhesion molecule-1; TNF-α: tumor necrosis factor-α; IL-12b: interleukin-12b; GAPDH: glyceraldehyde-3-phosphate dehydrogenase (internal reference).</p

Expression of NF-κB signaling pathway molecules in rat kidney tissues in different age groups.

<p>(<b>A</b>) Western blot detection of the expression of NF-κB signaling pathway proteins in various rat groups. (<b>B</b>) Quantitative analysis of gray scale. Compared with the 3-month-old group, * indicates P<0.05; compared with the 12-month-old group, # indicates P<0.05.</p

RT-qPCR detection of the expression of inflammatory cytokines in rat renal tissues in different age groups.

<p>Compared with the 3-month-old group, the fold change in inflammatory cytokines in the 12-month-old and 24-month-old groups  = 2^−ΔΔCT. Compared with the 3-month-old group, * indicates P<0.05; compared with the 12-month-old group, # indicates P<0.05.</p

Effect of diet on metabolic parameters and renal function in the aged rats.

<p>BMI, body mass index; CON, control animals; CR, calorie-restricted diet; HC, high-calorie diet. Data are presented as means ± SD (n  = 6), *<i>P</i><0.05 <i>vs.</i> CON.</p

Bnip3, Ambra1 decreased in HC kidneys and increased in CR kidneys compared with CON kidneys.

<p>The expression of Bnip3 and Ambra1was analyzed by Western blotting in the kidneys of CON, CR, and HC Fischer 344 rats. <b>a:</b> The expression of Bnip3 and Ambra1 was detected by Western blotting. <b>b:</b>Quantitative analysis of the band density for Bnip3. <b>c:</b>Quantitative analysis of the band density for Ambra1. The protein expression data are presented as the mean ± SD (n  = 6). *p<0.05 vs. CON.CON, control animals; CR, calorie-restricted diet; HC, high-calorie diet.</p

The level of 8-OHdG was increased in HC kidneys and decreased in CR kidneys.

<p>Immunohistochemistry staining results for LC3 proteins and 8-OHdG in the kidneys of the CON, CR and HC Fischer 344 rats were scanned by a microscope. CON, control animals; CR, calorie-restricted diet; HC, high-calorie diet.</p