The effects of polymicrobial sepsis with diabetes mellitus on kidney tissues in ovariectomized rats

Figures & data

TABLE 1.  Experimental protocol of rat groups

Groups		1st day	2–120th day	121–124th day	124–154th day	155th day	155th day and 20th hour
1	Sham	Waiting period					I.P. thiopental disodium 50 mg/kg
2	Ovariectomy	Ovariectomy surgery	Waiting period
3	Ovariectomy + diabetes	Ovariectomy surgery	Waiting period	Diabetes induction		Waiting period
4	Ovariectomy + sepsis	Ovariectomy surgery	Waiting period			Sepsis induction
5	Ovariectomy + diabetes + sepsis	Ovariectomy surgery	Waiting period	Diabetes induction	Waiting period	Sepsis induction

TABLE 2.  Differences between weights of the rats after waiting period

	Initial weight	Final weight	Difference (%)
Sham	221	261	18.10
Ovariectomy	215	279	29.77
Ovariectomy + diabetes	225	195	−13.33
Ovariectomy + sepsis	219	280	27.85
Ovariectomy + diabetes + sepsis	226	201	−11.06

TABLE 3.  The effects of ovariectomy on total glutathione (GSH), lipid peroxidation levels, enzymatic activities of LPO, catalase (CAT), and superoxide dismutase (SOD) of sepsis, diabetes, and sepsis and diabetes on kidney tissues of rats

Kidney tissues	N	CAT activity (mmol/min/mg tissue)	SOD activity (mmol/min/mg tissue)	Amount of LPO (nmol/g tissue)	Amount of GSH (nmol/mg tissue)
Sham	9	92.3 ± 0.5a	158.3 ± 0.5e	15.25 ± 0.15a	3.46 ± 0.04a
Ovariectomy	9	96.9 ± 0.4b	129.4 ± 0.6d	52.61 ± 0.84b	3.70 ± 0.02b
Ovariectomy + diabetes	9	99.3 ± 0.3c	102.6 ± 0.3c	86.93 ± 2.77d	4.84 ± 0.01e
Ovariectomy + sepsis	9	110.0 ± 1.0e	59.7 ± 0.2b	77.04 ± 0.22c	4.76 ± 0.01c
Ovariectomy + diabetes + sepsis	9	104.4 ± 1.0d	45.3 ± 0.7a	105.42 ± 0.37e	4.92 ± 0.01d

Notes: Means in the same column by the same letter are not significantly different to the one way ANOVA–Duncan test (α = 0.05). Results are means ± SE. N: Number of animals.

FIGURE 1. Blood glucose levels of the rats after waiting period.

*p< 0.05 when compared to sham group.

FIGURE 2. Serum levels of BUN (A) and creatinine (B) in experimental groups.

*p < 0.05 when compared to sham group.

FIGURE 3. Light microscopy of kidney in the control (A, B) and the ovariectomy group (C, D). gl, glomerulus; d, distal tubules; p, proximal tubules; t, collecting tubules; dye: hematoxylin–eosin; magnification bars: 40 μm.

FIGURE 4. Light microscopy of kidney in the diabetes (A, B) and sepsis group (C, D). gl, glomerulus; p, proximal tubules; asterisk, reduced glomerulus; underlined asterisk, inflammatory cell infiltrations at nodular form; arrow, proximal tubules with apical degenerations; arrow with double head, inflammatory cell infiltrations at diffuse form; asterisk with white filling, proximal tubule with integrity loss; arrow head, eosinophilic accumulations in intertubular area; dye: hematoxylin–eosin; magnification bars: 40 μm.

FIGURE 5. Light microscopy of kidney in the ovariectomy and diabetes (A, B) and ovariectomy and sepsis (C, D), and diabetes and sepsis group (E, F). gl, glomerulus; asterisk, glomerulus with segmental necrosis; double asterisks, inflammatory cell infiltrations at diffuse form; thick arrow, inflammatory cell infiltrations at nodular form; thin arrow, cells of distal tubules with hydropic degenerations ; arrow with double head and inset of E, proximal tubules with apical degenerations, asterisk with white filling, glomeruli which their capillaries were congested arrowhead, erythrocytes in inter-tubular area; dye: hematoxylin–eosin; magnification bars: 40 μm.

FIGURE 6. Light microscopy of kidney in the ovariectomy, sepsis and diabetes group (A–F). gl, glomerulus; sg, sclerotic glomeruli; gn, glomerulonephritis; asterisk, proteinous accumulations in inter-tubular area and urinary space; double asterisks, inflammatory cell infiltrations at diffuse form; underlined asterisk, proximal tubule with integrity loss; thick arrows, proliferating extra-mesangial cells in cortex; thin arrow, giant cells in cortex; inset of A, glomeruli with dilated capillary; inset of C, reduced glomerulus; inset of E, degenerated tubules; dye: hematoxylin–eosin; magnification bars: 40 μm.

FIGURE 7. Immunohistochemical staining of NF-κB (p65) in tissue sections of control (A, B), ovariectomy (C, D), sepsis (E, F), and diabetes (G, H) groups. gl, glomerulus; d, distal tubules; p, proximal tubules; t, collecting tubules; positivity was seen in especially cytoplasmic pattern of tubular cells in ovariectomy (C, D), sepsis (E, F; the most expression of p65), and diabetes (G, H) groups; magnification bars: 30 μm.

FIGURE 8. Immunohistochemical staining of NF-κB (p65) in tissue sections of ovariectomy–diabetes (A, B), ovariectomy–sepsis (C, D), and diabetes–sepsis (E, F) groups. gl, glomerulus; d, distal tubules; p, proximal tubules; t, collecting tubules; arrows indicate extramesangial cells with p65 positive nuclei and cytoplasm; positivity was seen in especially cytoplasmic pattern of tubular cells, both cytoplasmic and nuclear pattern of extramesangial cells and rarely in glomeruli. The most expression of p65 was found in diabetes–sepsis group; magnification bars: 30 μm.

FIGURE 9. Immunohistochemical staining of NF-κ B (p65) in tissue sections of ovariectomy–diabetes and sepsis group (A–D). gl, glomerulus; d, distal tubules; p, proximal tubules; t, collecting tubules; arrows indicate extramesangial cells with p65 positive nuclei and cytoplasm; positivity was seen in especially cytoplasmic pattern of tubular cells, both cytoplasmic and nuclear pattern of extramesangial cells and commonly in glomeruli. The highest expression of p65 was found in this group among the all groups; magnification bars: 30 μm.