Role of Fucoxanthin towards Cadmium-induced renal impairment with the antioxidant and anti-lipid peroxide activities

Figures & data

Table 1. Housekeeping Gene (mouse β-actin) and targeted gene expression primer sequences

Gene	Primer sequence	PCR amplicon size
M-β-actin-F	GTGCTATGTTGCTCTAGACTTCG	174bp
M-β -actin-R	ATGCCACAGGATTCCATACC
M-ERK1-F	GGCTTTCTGACGGAGTATGTG	197bp
M-ERK1-R	GGGGAACCCAAGATACCTAGA
M- ERK2-F	GGTTGTTCCCAAATGCTGAC	138bp
M- ERK2-R	GCTCATCACTTGGGTCATAATACT
M- MEK1-F	TGACGCAGAAGCAGAAGGTG	90bp
M-MEK1-R	TGAAGACCACTCCACCGTTG
M-eIf2a-F	TAATCAATGTCGCTAACAAGGG	152bp
M-eIf2a-R	AAGTTGTAGGTTAGGCGTCCC
M-p-eIf2a-F	TTCTACAGAAACCATGCCCAT	123bp
M-p-eIf2a-R	TTGATAACTGCCATAGCCTGAT
M-GRP78-F	GCCAACTGTAACAATCAAGGTCT	234bp
M-GRP78-R	TCAGGTGTCAGGCGGTTTT
M-GRP94-F	AGGTGTTGTGGATTCCGATGA	225bp
M-GRP94-R	AGTTTAGCAAGCCGTGTTCG
M-caspase3-F	TGACTGGAAAGCCGAAACTCT	213bp
M-caspase3-R	TGCTGCAAAGGGACTGGAT
M-caspase8-F	TCAAAGTGCCCTTCCCTGT	197bp
M-caspase8-R	TTCTTCACCGTAGCCATTCC
M-caspase9-F	AGAACGACCTGACTGCCAAGA	101bp
M-caspase9-R	ATGAGAGAGGATGACCACCACA

M: mouse; F: forward; R: reverse; ERK1:extracellular signal-regulated kinase 1; MEK1, also named MAP2K1, mitogen-activated protein kinase1; eIF2a, Eukaryotic translation initiation factor 2A;p-IF2a, phosphorylated eukaryotic translation initiation factor2a;GRP78, glucose regulatory protein 78.

Table 2. Renal cadmium concentration

Group	Control	NCG	PCG	F1	F2	F3
Cadmium concentration (ng/ml)	5.28 ±0.45	128.55 ±6.75*	92.81 ±3.24*#	120.98 ±9.55*^	79.49 ±6.31*#^	75.49 ±4.56*#^

Single factor analysis of variance between multiple groups was performed using Duncan methods. ‘*’ compared with the control group, P < 0.05; ‘#’ compared with the negative control group, P < 0.05; ‘^’ compared with the positive control group, P < 0.05. Control group: no CdCl₂ administration; NCG, negative control group, only CdCl₂ orally at a dose of 30 milligram (mg)/ kilogram (kg) body weight (bw)/day for 30 days; PCG, positive control group, CdCl₂+ Shenfukang tablets orally at a dose of 50 mg/kg bw/day for 14 days; F1, CdCl₂+ low fucoxanthin concentration group, mice received fucoxanthin orally at a dose of 10 mg/kg bw/day for 14 days; F2, CdCl₂+ mice received fucoxanthin orally at a dose of 25 mg/kg bw/day for 14 days; F3, CdCl₂+ mice received fucoxanthin orally at a dose of 50 mg/kg bw/day for 14 days.

Figure 1. Hematoxylin and eosin (H &E) staining of renal in CdCl₂ exposure after the treatment with fucoxanthin and Shenfukang tablets supplementation (1) the control group: no CdCl₂ administration; (2) NCG, negative control group, only CdCl₂ treatment; (3) PCG, positive control group, CdCl₂+ Shenfukang tablets; (4) F1, CdCl₂ + 10 mg/kg fucoxanthin body weight for 14 day treatment ; (5) F2, CdCl₂ + 25 mg/kg body weight fucoxanthin; (6) F3,CdCl₂ + 50 mg/kg body weight fucoxanthin. Each group used 20 mice

Figure 2. The quantity of the cross-sectional area of the glomeruli (μm²) Single factor analysis of variance between multiple groups was performed using Duncan methods. ‘*’ compared with the control group, P < 0.05; ‘#’ compared with the negative control group, P < 0.05; ‘^’ compared with the positive control group, P < 0.05. Each group used 20 mice

Figure 3. The semi-quantitative score of renal tubular damage in mice Single factor analysis of variance between multiple groups was performed using Duncan methods. ‘*’ compared with the control group, P < 0.05; ‘#’ compared with the negative control group, P < 0.05. Each group used 20 mice

Figure 4. Ultrastructural changes after treatment with fucoxanthin and Shenfukang tablets administration (1) Control group, (2) NCG, (3) PCG, (4) F3, (a) mitochondria damage, (b) karyopyknosis, and (c) mitochondrial rupture. Each group used 20 mice

Figure 5. Semi-quantitative evaluation of mitochondrial damage in mouse kidney cells Single factor analysis of variance between multiple groups was performed using Duncan methods.‘*’ compared with the control group, P < 0.05; ‘#’ compared with the negative control group, P < 0.05. Each group used 20 mice

Table 3. Changes in the BUN, KIM-1 and NGAL levels

	Control	NCG	PCG	F1	F2	F3
BUN (mmol/L)	2.70 ±0.12	4.13 ±0.16*	2.94 ±0.11#	4.35 ±0.20*^	2.64 ±0.16#	2.55 ±0.20#
KIM-1 (pg/ml)	67.67 ±35.69	713.86 ±117.82*	473.67 ±97.94*#	404.92 ±73.13*#	174.85 ±87.75#^	281.07 ±79.56*#^
NGAL (ng/ml)	39.61 ±9.66	95.52 ±9.38*	45.77 ±9.80#	76.70 ±13.19*#^	63.60 ±6.08*#^	60.85 ±12.76*#^

Single factor analysis of variance between multiple groups was performed using Duncan methods. ‘*’ compared with the control group, P < 0.05; ‘#’ compared with the negative control group, P < 0.05; ‘^’ compared with the positive control group, P < 0.05. Control group: no CdCl₂ administration; NCG, negative control group, only CdCl₂ orally at a dose of 30 milligram (mg)/ kilogram (kg) body weight (bw) /day for 30 days; PCG, positive control group, CdCl₂+ Shenfukang tablets orally at a dose of 50 mg/kg bw/day for 14 days; F1, CdCl₂+ fucoxanthin orally at a dose of 10 mg/kg bw/day for 14 days; F2, CdCl₂+ fucoxanthin orally at a dose of 25 mg/kg bw/day for 14 days; F3, CdCl₂+ fucoxanthin orally at a dose of 50 mg/kg bw/day for 14 days.

Figure 6. Changes in Renal POD, SOD, CAT and APX Activities (U/g) Single factor analysis of variance between multiple groups was performed using Duncan methods. ‘*’ compared with the control group, P < 0.05; ‘#’ compared with the negative control group, P < 0.05; ‘^’ compared with the positive control group, P < 0.05. Each group used 20 mice

Figure 7. Effect of fucoxanthin on the expression of apoptosis-associated genes in the cadmium kidney-damaged mouse model Single factor analysis of variance between multiple groups was performed using Duncan methods.‘*’ compared with the control group, P < 0.05; ‘#’ compared with the negative control group, P < 0.05. Each group used 20 mice