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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

The effect of nuciferine or siRNA PASK on TG accumulation in HepG2 cells of OA-induced hepatic steatosis. (A) TG concentration tested by ELISA. (B,C) Oil-red-O staining. Original magnification was 200×, scale bars represent 75 μm. SiRNA PASK HepG2 cells and HepG2 cells incubated with increasing concentrations of nuciferine (NF: 10, 25, and 50 μM) or vitamin E (25 μM) both treated with OA (40 μM). The measurement was described in the Section “Materials and Methods”. Values are Mean ± SEM of three independent experiments performed in triplicates. Significant differences with OA group were designated as ∗∗P < 0.01 and ∗∗∗P < 0.001. (Control, siRNA scrambled; NF, Nuciferine.)
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Figure 3: The effect of nuciferine or siRNA PASK on TG accumulation in HepG2 cells of OA-induced hepatic steatosis. (A) TG concentration tested by ELISA. (B,C) Oil-red-O staining. Original magnification was 200×, scale bars represent 75 μm. SiRNA PASK HepG2 cells and HepG2 cells incubated with increasing concentrations of nuciferine (NF: 10, 25, and 50 μM) or vitamin E (25 μM) both treated with OA (40 μM). The measurement was described in the Section “Materials and Methods”. Values are Mean ± SEM of three independent experiments performed in triplicates. Significant differences with OA group were designated as ∗∗P < 0.01 and ∗∗∗P < 0.001. (Control, siRNA scrambled; NF, Nuciferine.)

Mentions: To examine the catabatic role of nuciferine and silenced PASK in TG accumulation, any decrease in TG content was evaluated using the ELISA kit and oil-red-O staining. Owing to the results of all parameters in the control group were almost consistent with the siRNA scrambled (data were not shown), for more concise, we selected siRNA scrambled as control in our next results. It was found that TG accumulated in OA treated cells as compared with the control (∗∗∗P < 0.001). The three concentrations of nuciferine decreased TG accumulation by 16.6, 33.7, and 52.7% respectively (∗∗P < 0.01 and ∗∗∗P < 0.001 vs. OA), while vitamin E (25 μM) and siRNA PASK reduced the TG content by 38.6% (∗∗P < 0.01 vs. OA) and 53% (∗∗∗P < 0.001 vs. OA) respectively as compared with OA group (Figure 3A). Microscopic examination using oil-red-O staining showed that lipid droplets in cells treated with 10, 25, and 50 μM nuciferine decreased by 15, 45.5, and 60.7% respectively as determined by MOD (∗∗∗P < 0.001 vs. OA), 54.6% for 25 μM vitamin E (∗∗∗P < 0.001 vs. OA) and 63% for siRNA PASK (∗∗∗P < 0.001 vs. OA) as compared with OA group (∗∗∗P < 0.001 vs. control) (Figures 3B,C). These data initially imply that both nuciferine and siRNA PASK had a regulatory effect on lipid 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)

The effect of nuciferine or siRNA PASK on TG accumulation in HepG2 cells of OA-induced hepatic steatosis. (A) TG concentration tested by ELISA. (B,C) Oil-red-O staining. Original magnification was 200×, scale bars represent 75 μm. SiRNA PASK HepG2 cells and HepG2 cells incubated with increasing concentrations of nuciferine (NF: 10, 25, and 50 μM) or vitamin E (25 μM) both treated with OA (40 μM). The measurement was described in the Section “Materials and Methods”. Values are Mean ± SEM of three independent experiments performed in triplicates. Significant differences with OA group were designated as ∗∗P < 0.01 and ∗∗∗P < 0.001. (Control, siRNA scrambled; NF, Nuciferine.)
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Related In: Results  -  Collection

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Figure 3: The effect of nuciferine or siRNA PASK on TG accumulation in HepG2 cells of OA-induced hepatic steatosis. (A) TG concentration tested by ELISA. (B,C) Oil-red-O staining. Original magnification was 200×, scale bars represent 75 μm. SiRNA PASK HepG2 cells and HepG2 cells incubated with increasing concentrations of nuciferine (NF: 10, 25, and 50 μM) or vitamin E (25 μM) both treated with OA (40 μM). The measurement was described in the Section “Materials and Methods”. Values are Mean ± SEM of three independent experiments performed in triplicates. Significant differences with OA group were designated as ∗∗P < 0.01 and ∗∗∗P < 0.001. (Control, siRNA scrambled; NF, Nuciferine.)
Mentions: To examine the catabatic role of nuciferine and silenced PASK in TG accumulation, any decrease in TG content was evaluated using the ELISA kit and oil-red-O staining. Owing to the results of all parameters in the control group were almost consistent with the siRNA scrambled (data were not shown), for more concise, we selected siRNA scrambled as control in our next results. It was found that TG accumulated in OA treated cells as compared with the control (∗∗∗P < 0.001). The three concentrations of nuciferine decreased TG accumulation by 16.6, 33.7, and 52.7% respectively (∗∗P < 0.01 and ∗∗∗P < 0.001 vs. OA), while vitamin E (25 μM) and siRNA PASK reduced the TG content by 38.6% (∗∗P < 0.01 vs. OA) and 53% (∗∗∗P < 0.001 vs. OA) respectively as compared with OA group (Figure 3A). Microscopic examination using oil-red-O staining showed that lipid droplets in cells treated with 10, 25, and 50 μM nuciferine decreased by 15, 45.5, and 60.7% respectively as determined by MOD (∗∗∗P < 0.001 vs. OA), 54.6% for 25 μM vitamin E (∗∗∗P < 0.001 vs. OA) and 63% for siRNA PASK (∗∗∗P < 0.001 vs. OA) as compared with OA group (∗∗∗P < 0.001 vs. control) (Figures 3B,C). These data initially imply that both nuciferine and siRNA PASK had a regulatory effect on lipid 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