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Nuciferine downregulates Per-Arnt-Sim kinase expression during its alleviation of lipogenesis and inflammation on oleic acid-induced hepatic steatosis in HepG2 cells.

Zhang DD, Zhang JG, Wu X, Liu Y, Gu SY, Zhu GH, Wang YZ, Liu GL, Li XY - Front Pharmacol (2015)

Bottom Line: It was found that nuciferine and silenced-PASK (siRNA PASK) both inhibited triglyceride (TG) accumulation and was effective in decreasing fatty acid (FFAs).In addition, nuciferine decreased TNF-a, IL-6 and IL-8 as well as the siRNA PASK group.Furthermore, the modulated effect of nuciferine on (OA)-induced HepG2 cells lipogenesis and inflammation, which was accompanied with PASK inhibition, was also consistent with siRNA PASK, implying that PASK might play a role in nuciferine-mediated regulation on NAFLD.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Pharmacy, Shanghai General Hospital, Shanghai Jiaotong University School of Medicine Shanghai, China.

ABSTRACT
Non-alcoholic fatty liver disease (NAFLD) is a prevalent liver disease associated with lipotoxicity, lipid peroxidation, oxidative stress, and inflammation. Nuciferine, an active ingredient extracted from the natural lotus leaf, has been reported to be effective for the prevention and treatment of NAFLD. Per-Arnt-Sim kinase (PASK) is a nutrient responsive protein kinase that regulates lipid and glucose metabolism, mitochondrial respiration, and gene expression. The aim of the present study was to investigate the protective effect of nuciferine against NAFLD and its inhibitory effect on PASK, exploring the possible underlying mechanism of nuciferine-mediated inhibition on NAFLD. Relevant biochemical parameters (lipid accumulation, extent of oxidative stress and release of inflammation cytokines) in oleic acid (OA)-induced HepG2 cells that mimicked steatosis in vitro were measured and compared with the control. It was found that nuciferine and silenced-PASK (siRNA PASK) both inhibited triglyceride (TG) accumulation and was effective in decreasing fatty acid (FFAs). The content of total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) were increased respectively by nuciferine and siRNA PASK without increase in glutathione (GSH). Malondialdehyde (MDA) was decreased respectively by nuciferine and siRNA PASK. In addition, nuciferine decreased TNF-a, IL-6 and IL-8 as well as the siRNA PASK group. IL-10 was increased by nuciferine and siRNA PASK respectively. Further investigation revealed that nuciferine and siRNA PASK could respectively regulate the expression of target genes involved in lipogenesis and inflammation, suggesting that nuciferine may be a potential therapeutic treatment for NAFLD. Furthermore, the modulated effect of nuciferine on (OA)-induced HepG2 cells lipogenesis and inflammation, which was accompanied with PASK inhibition, was also consistent with siRNA PASK, implying that PASK might play a role in nuciferine-mediated regulation on NAFLD.

No MeSH data available.


Related in: MedlinePlus

Oleic acid-induced TG accumulation in HepG2 cells. (A) The Dose dependent effect of OA (40, 80, 120, 160, and 200 μM) on cytotoxicity in HepG2 cells. (B,C) The effect of OA (40, 80, 120, 160, and 200 μM) on TG accumulation in HepG2 cells. Original magnification was 200×, scale bars represent 75 μm. Values are Mean ± SEM of three independent experiments performed in triplicates. Statistically significant at ∗P < 0.05, ∗∗P < 0.01 and ∗∗∗P < 0.001 compared with the control.
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Figure 1: Oleic acid-induced TG accumulation in HepG2 cells. (A) The Dose dependent effect of OA (40, 80, 120, 160, and 200 μM) on cytotoxicity in HepG2 cells. (B,C) The effect of OA (40, 80, 120, 160, and 200 μM) on TG accumulation in HepG2 cells. Original magnification was 200×, scale bars represent 75 μm. Values are Mean ± SEM of three independent experiments performed in triplicates. Statistically significant at ∗P < 0.05, ∗∗P < 0.01 and ∗∗∗P < 0.001 compared with the control.

Mentions: To mimic the hepatic steatosis in vitro, HepG2 cells were treated with 0–200 μM concentrations of OA for 24 h to induce TG steatosis. The optimal concentration depended on the cytotoxic effect of OA and the TG content evaluated by oil-red-O staining. Compare to the control (HepG2 cells without OA), the cell viability value ranged from 52 to 91% (Figure 1A) (∗P < 0.05, ∗∗P < 0.01 and ∗∗∗P < 0.001) while the recovered oil-red-O content (determined by MOD) was increased significantly (Figures 1B,C) (∗∗P < 0.01 and ∗∗∗P < 0.001). Cell viability values greater than 90% were considered the unaffected concentration combined with the increased TG content. Therefore, 40 μM OA was selected as the final concentration to induce TG accumulation.


Nuciferine downregulates Per-Arnt-Sim kinase expression during its alleviation of lipogenesis and inflammation on oleic acid-induced hepatic steatosis in HepG2 cells.

Zhang DD, Zhang JG, Wu X, Liu Y, Gu SY, Zhu GH, Wang YZ, Liu GL, Li XY - Front Pharmacol (2015)

Oleic acid-induced TG accumulation in HepG2 cells. (A) The Dose dependent effect of OA (40, 80, 120, 160, and 200 μM) on cytotoxicity in HepG2 cells. (B,C) The effect of OA (40, 80, 120, 160, and 200 μM) on TG accumulation in HepG2 cells. Original magnification was 200×, scale bars represent 75 μm. Values are Mean ± SEM of three independent experiments performed in triplicates. Statistically significant at ∗P < 0.05, ∗∗P < 0.01 and ∗∗∗P < 0.001 compared with the control.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4612658&req=5

Figure 1: Oleic acid-induced TG accumulation in HepG2 cells. (A) The Dose dependent effect of OA (40, 80, 120, 160, and 200 μM) on cytotoxicity in HepG2 cells. (B,C) The effect of OA (40, 80, 120, 160, and 200 μM) on TG accumulation in HepG2 cells. Original magnification was 200×, scale bars represent 75 μm. Values are Mean ± SEM of three independent experiments performed in triplicates. Statistically significant at ∗P < 0.05, ∗∗P < 0.01 and ∗∗∗P < 0.001 compared with the control.
Mentions: To mimic the hepatic steatosis in vitro, HepG2 cells were treated with 0–200 μM concentrations of OA for 24 h to induce TG steatosis. The optimal concentration depended on the cytotoxic effect of OA and the TG content evaluated by oil-red-O staining. Compare to the control (HepG2 cells without OA), the cell viability value ranged from 52 to 91% (Figure 1A) (∗P < 0.05, ∗∗P < 0.01 and ∗∗∗P < 0.001) while the recovered oil-red-O content (determined by MOD) was increased significantly (Figures 1B,C) (∗∗P < 0.01 and ∗∗∗P < 0.001). Cell viability values greater than 90% were considered the unaffected concentration combined with the increased TG content. Therefore, 40 μM OA was selected as the final concentration to induce TG accumulation.

Bottom Line: It was found that nuciferine and silenced-PASK (siRNA PASK) both inhibited triglyceride (TG) accumulation and was effective in decreasing fatty acid (FFAs).In addition, nuciferine decreased TNF-a, IL-6 and IL-8 as well as the siRNA PASK group.Furthermore, the modulated effect of nuciferine on (OA)-induced HepG2 cells lipogenesis and inflammation, which was accompanied with PASK inhibition, was also consistent with siRNA PASK, implying that PASK might play a role in nuciferine-mediated regulation on NAFLD.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Pharmacy, Shanghai General Hospital, Shanghai Jiaotong University School of Medicine Shanghai, China.

ABSTRACT
Non-alcoholic fatty liver disease (NAFLD) is a prevalent liver disease associated with lipotoxicity, lipid peroxidation, oxidative stress, and inflammation. Nuciferine, an active ingredient extracted from the natural lotus leaf, has been reported to be effective for the prevention and treatment of NAFLD. Per-Arnt-Sim kinase (PASK) is a nutrient responsive protein kinase that regulates lipid and glucose metabolism, mitochondrial respiration, and gene expression. The aim of the present study was to investigate the protective effect of nuciferine against NAFLD and its inhibitory effect on PASK, exploring the possible underlying mechanism of nuciferine-mediated inhibition on NAFLD. Relevant biochemical parameters (lipid accumulation, extent of oxidative stress and release of inflammation cytokines) in oleic acid (OA)-induced HepG2 cells that mimicked steatosis in vitro were measured and compared with the control. It was found that nuciferine and silenced-PASK (siRNA PASK) both inhibited triglyceride (TG) accumulation and was effective in decreasing fatty acid (FFAs). The content of total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) were increased respectively by nuciferine and siRNA PASK without increase in glutathione (GSH). Malondialdehyde (MDA) was decreased respectively by nuciferine and siRNA PASK. In addition, nuciferine decreased TNF-a, IL-6 and IL-8 as well as the siRNA PASK group. IL-10 was increased by nuciferine and siRNA PASK respectively. Further investigation revealed that nuciferine and siRNA PASK could respectively regulate the expression of target genes involved in lipogenesis and inflammation, suggesting that nuciferine may be a potential therapeutic treatment for NAFLD. Furthermore, the modulated effect of nuciferine on (OA)-induced HepG2 cells lipogenesis and inflammation, which was accompanied with PASK inhibition, was also consistent with siRNA PASK, implying that PASK might play a role in nuciferine-mediated regulation on NAFLD.

No MeSH data available.


Related in: MedlinePlus