Nutr Res Pract.  2024 Oct;18(5):633-646. 10.4162/nrp.2024.18.5.633.

Cladophora glomerata Kützing extract exhibits antioxidant, anti-inflammation, and anti-nitrosative stress against impairment of renal organic anion transport in an in vivo study

• Ontawong A
• Aida CJ
• Vivithanaporn P
• Amornlerdpison D
• Vaddhanaphuti CS

• KMID: 2560049
• DOI: http://doi.org/10.4162/nrp.2024.18.5.633

Abstract

BACKGROUND/OBJECTIVES
Cladophora glomerata extract (CGE), rich in polyphenols, was reported to exhibit antidiabetic and renoprotective effects by modulating the functions of protein kinases-mediated organic anion transporter 1 (Oat1) and 3 (Oat3) in rats with type 2 diabetes mellitus (T2DM). Nevertheless, the antioxidant effects of CGE on such renoprotection have not been investigated. This study examined the mechanisms involved in the antioxidant effects of CGE on renal organic anion transport function in an in vivo study.
MATERIALS/METHODS
Diabetes was induced in the rats through a high-fat diet combined with a single dose of 40 mg/kg body weight (BW) streptozotocin. Subsequently, normal-diet rats were supplemented with a vehicle or 1,000 mg/kg BW of CGE, while T2DM rats were supplemented with a vehicle, CGE, or 200 mg/kg BW of vitamin C for 12 weeks. The study evaluated the general characteristics of T2DM and renal oxidative stress markers. The renal organic transport function was assessed by measuring the para-aminohippurate (PAH) uptake using renal cortical slices and renal inflammatory cytokine expression in the normal diet (ND) and ND + CGE treated groups.
RESULTS
CGE supplementation significantly reduced hyperglycemia, hypertriglyceridemia, insulin resistance, and renal lipid peroxidation in T2DM rats. This was accompanied by the normalization of high expressions of renal glutathione peroxidase and nuclear factor kappa B by CGE and vitamin C. The renal anti-inflammation of CGE was evidenced by the reduction of tumor necrosis factor-1α and interleukin-1β. CGE directly blunted sodium nitroprusside-induced renal oxidative/nitrosative stresses and mediated the PAH uptake in the normally treated CGE in rats was particularly noteworthy. These data also correlated with reduced nitric oxide production, highlighting the potential of CGE as a therapeutic agent for managing T2DM-related renal complications.
CONCLUSION
These findings suggest that CGE has antidiabetic effects and directly prevents diabetic nephropathy through oxidative/nitrosative stress pathways.

Keyword

Antioxidants; nitrosative stress; organic anion transporter; oxidative stress; type 2 diabetes mellitus

Figure

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  Fig. 1 Effects of CGE on the plasma total, reduced, and oxidized GSH levels, determined in plasma samples from each experimental group using a colorimetric assay kit. The results are expressed as the mean ± SE of the mean (n = 6).CGE, Cladophora glomerata extract; T2DM or DM, type 2 diabetes mellitus; ND, normal diet; VC, vitamin C; GSH, glutathione.

• 

  Fig. 2 Effects of CGE on renal cortical MDA level, determined in renal cortical tissue homogenates from each experimental group using a commercial TBARS assay kit. The results are expressed as mean ± SE of the mean (n = 6).CGE, Cladophora glomerata extract; T2DM or DM, type 2 diabetes mellitus; ND, normal diet; VC, vitamin C; MDA, malondialdehyde.*P < 0.05 indicates a significant difference from ND; #P < 0.05 indicates a significant difference from DM rats.

• 

  Fig. 3 Effect of CGE on the expression of stress-sensitive markers. (A) p65NF-κB expression in subcellular fractions of rat kidneys. Anti-p65NF-κB antibody was then detected, whereas anti-β-actin and anti-lamin B1 antibodies were used as a loading control and nuclei marker, respectively. The data are expressed as mean ± SE of the mean and repeated for separate sets of animals (n = 3). p65NF-κB protein expression is shown at the top, and the quantification of relative protein expression in each fraction is at the bottom. (B) Nrf2 expression in the subcellular fractions of the rat kidneys. Anti-Nrf2 antibody was subsequently detected, whereas anti-β-actin and anti-lamin B1 antibodies were used as a loading control and nuclei marker, respectively. The data are expressed as the mean ± SE of the mean and repeated for separate sets of animals (n = 3). Nrf2 protein expression is shown at the top and quantification of the relative protein expression in each fraction at the bottom. (C) mRNA expression levels of CAT, GPx, and Cu-Zn SOD from experimental rat kidneys. The values represent mean ± SE of the mean (n = 6).CGE, Cladophora glomerata extract; T2DM or DM, type 2 diabetes mellitus; ND, normal diet; VC, vitamin C; p65NF-κB, p65 nuclear factor κB; Nrf2, nuclear factor (erythroid-derived 2)-like 2; CAT, catalase; GPx, glutathione peroxidase; Cu-Zn SOD, copper-zinc superoxide dismutase.*P < 0.05 indicates the significant differences from ND; #P < 0.05 indicates significant differences from DM rats.

• 

  Fig. 4 Effect of CGE on the protein expressions of pro-inflammatory cytokines. (A) TNF-α and (B) IL-1β expression in whole-cell lysate from the renal cortical tissues in ND and ND + CGE groups. Anti-TNF-α and anti-IL-1β antibodies were then detected, whereas anti-β-actin antibody was used as a loading control. The values represent the mean ± SE of the mean from separate sets of animals (n = 4).CGE, Cladophora glomerata extract; ND, normal diet; TNF-α, tumor necrosis factor-alpha; IL-1β, interleukin-1β.*P < 0.05 indicates significant differences from ND rats.

• 

  Fig. 5 Effect of CGE on sodium nitroprusside-induced NO production in renal cortical tissue. (A) PAH transport in the renal cortical slices from ND and ND + CGE rats. Rat renal cortical slices were incubated for 30 min in the buffer containing 10 μM of tritiated PAH in the presence or absence of SNP. Data are expressed as mean ± SE of the mean (n = 5). (B) Sodium nitroprusside induced NO production in renal cortical tissues from ND and ND + CGE rats. Rat renal cortical slices were incubated for 30 min in the buffer with or without SNP (n = 5). The data are expressed as mean ± SE.CGE, Cladophora glomerata extract; ND, normal diet; NO, nitric oxide; PAH, para-aminohippurate; SNP, sodium nitroprusside; T/M, tissue to medium.*P < 0.05 indicates significant differences from slices incubated with buffer alone.

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