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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

Cytotoxicity and apoptosis of nuciferine. (A) Cytotoxicity of nuciferine in HepG2 cells. HepG2 cells were incubated with OA (40 μM) and increased concentrations of nuciferine (10, 25, 50, 100, 250, and 500 μM) for 24 h, and cell viability was measured by CCK8 assay. (B) The dose dependent effect of nuciferine (10, 25, and 50 μM) on cell apoptosis in HepG2 cells. Values are Mean ± SEM of three independent experiments performed in triplicates. Statistically significant at ∗∗∗P < 0.001 compared with the control.
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Figure 2: Cytotoxicity and apoptosis of nuciferine. (A) Cytotoxicity of nuciferine in HepG2 cells. HepG2 cells were incubated with OA (40 μM) and increased concentrations of nuciferine (10, 25, 50, 100, 250, and 500 μM) for 24 h, and cell viability was measured by CCK8 assay. (B) The dose dependent effect of nuciferine (10, 25, and 50 μM) on cell apoptosis in HepG2 cells. Values are Mean ± SEM of three independent experiments performed in triplicates. Statistically significant at ∗∗∗P < 0.001 compared with the control.

Mentions: To eliminate any misinterpretation owing to cell apoptosis or death caused by toxicity of nuciferine, apoptosis, and cytotoxicity on HepG2 cells were assessed. Cells were treated with the OA (40 μM) and 0–500 μM nuciferine for 24 h. The cytotoxic effect of nuciferine was determined by cell viability. Compare to the control (HepG2 cells without both OA and nuciferine), the value ranged from 57 to 91% at 0–500 μM concentrations and values greater than 85% were considered unaffected concentrations (∗∗∗P < 0.001 vs. control). Therefore, 10 μM, 25 μM, and 50 μM nuciferine concentrations were used to reduce fatty liver (Figure 2A). In addition, nuciferine concentrations ranging from 10 to 50 μM did not significantly induce apoptosis of HepG2 cells according to flow cytometric analysis (Figure 2B).


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)

Cytotoxicity and apoptosis of nuciferine. (A) Cytotoxicity of nuciferine in HepG2 cells. HepG2 cells were incubated with OA (40 μM) and increased concentrations of nuciferine (10, 25, 50, 100, 250, and 500 μM) for 24 h, and cell viability was measured by CCK8 assay. (B) The dose dependent effect of nuciferine (10, 25, and 50 μM) on cell apoptosis in HepG2 cells. Values are Mean ± SEM of three independent experiments performed in triplicates. Statistically significant at ∗∗∗P < 0.001 compared with the control.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 2: Cytotoxicity and apoptosis of nuciferine. (A) Cytotoxicity of nuciferine in HepG2 cells. HepG2 cells were incubated with OA (40 μM) and increased concentrations of nuciferine (10, 25, 50, 100, 250, and 500 μM) for 24 h, and cell viability was measured by CCK8 assay. (B) The dose dependent effect of nuciferine (10, 25, and 50 μM) on cell apoptosis in HepG2 cells. Values are Mean ± SEM of three independent experiments performed in triplicates. Statistically significant at ∗∗∗P < 0.001 compared with the control.
Mentions: To eliminate any misinterpretation owing to cell apoptosis or death caused by toxicity of nuciferine, apoptosis, and cytotoxicity on HepG2 cells were assessed. Cells were treated with the OA (40 μM) and 0–500 μM nuciferine for 24 h. The cytotoxic effect of nuciferine was determined by cell viability. Compare to the control (HepG2 cells without both OA and nuciferine), the value ranged from 57 to 91% at 0–500 μM concentrations and values greater than 85% were considered unaffected concentrations (∗∗∗P < 0.001 vs. control). Therefore, 10 μM, 25 μM, and 50 μM nuciferine concentrations were used to reduce fatty liver (Figure 2A). In addition, nuciferine concentrations ranging from 10 to 50 μM did not significantly induce apoptosis of HepG2 cells according to flow cytometric analysis (Figure 2B).

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