Kaempferol suppresses acetaminophen-induced liver damage by upregulation/activation of SIRT1

Figures & data

Table 1. Antibodies used in this study.

Target	Cat. no.	Manufacturer	Dilution	kDa
SIRT1	sc-74504	Santa Cruz Biotechnology	1:1000	120
PARP1	sc-8007	Santa Cruz Biotechnology	1:1000	110
CYP2E1	ab28146	Abcam	1:1000	50
Bax	2772	Call Signaling Technology	1:1000	20
Bcl-2	Sc-7382	Santa Cruz Biotechnology	1:1000	26
Cleaved caspase-3	9661	Cell Signaling Technology	1:500	19
MnSOD	ab13533	Abcam	1:1000	26
NF-κB p65	ab16502	Abcam	1:1000	65
P53	Sc-126	Santa Cruz Biotechnology	1:1000	53
FOXO-1	2880	Cell Signaling Technology	1:1000	80
Lamin B1	ab65986	Abcam	1:1000	70
β-actin	4967	Cell Signaling Technology	1:2000	45
Acetyl lysine	9441	Cell Signaling Technology	1:500

Figure 1. Circulatory levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and γ-glutamyl transferase (γ-GT). Serum levels of ALT (A), AST (B) and γ-GT (C) in the experimental groups of rats were determined using an automatic-analyser. Values are expressed as mean ± standard deviation (SD; eight rats/group). ^avs. control group. ^bvs. kaempferol-treated control group. ^cvs. APAP (acetaminophen)-treated rats.

Figure 2. Levels of tumour necrosis factor-α (TNF-α) (A) and interleukin 6 (IL-6) (B) in the liver homogenates of the experimental groups. Values are expressed as mean ± SD (eight rats/group). ^avs. control group. ^bvs. kaempferol-treated control group. ^cvs. APAP (acetaminophen)-treated rats.

Figure 3. Levels of malondialdehyde (MDA) (A), reactive oxygen species (ROS) (B), glutathione (GSH) (C) and superoxide dismutase (SOD) (D) in the liver homogenates of the experimental groups. Values are expressed as mean ± SD (eight rats/group). ^avs. control group. ^bvs. kaempferol-treated control group. ^cvs. APAP (acetaminophen)-treated rats.

Figure 4. Light micrographs of liver tissues from all groups after H&E staining. (A, B) control and kaempferol-treated rats, respectively, showing normal hepatocytes (H) and blood sinusoids (S) around the central veins (CVs). (C) APAP-treated rats showing swollen hepatocytes and wide blood sinusoids around the CVs. Some apoptotic and necrotic hepatocytes (H1), and inflammatory cells (arrows) were also seen. Some accumulation of erythrocytes was observed inside the CVs. (D) Kaempferol + APAP-treated rats showing normal hepatocytes and blood sinusoids around the CVs. Very few inflammatory cells (arrows) were seen around the CVs.

Figure 5. Protein levels of Bcl-2 (A), Bax (B), cleaved caspase-3 (C) and manganese superoxide dismutase (MnSOD) (D) in the liver homogenates of all experimental groups. Values are expressed as mean ± SD (six rats/group). ^avs. control group. ^bvs. kaempferol-treated control group. ^cvs. APAP (acetaminophen)-treated rats.

Figure 6. Nuclear activity of SIRT1 (A), protein levels of SIRT1 (B), cytochrome-2E1 (CYP2E1) (C) and PARP1 (D) in the liver homogenates of all experimental groups. Values are expressed as mean ± SD (six rats/group). ^avs. control group. ^bvs. kaempferol-treated control group. ^cvs. APAP (acetaminophen)-treated rats.

Figure 7. Total nuclear levels of NF-κB P65/acetylated NF-κB p65 (A), P53/acetylated p53 (B), FOXO-1/acetylated FOXO-1 (C) in the livers of all experimental groups. Values are expressed as mean ± SD (six rats/group). ^avs. control group. ^bvs. kaempferol-treated control group. ^cvs. APAP (acetaminophen)-treated rats.