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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).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.

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 cell lipid peroxidation in HepG2 cells of OA-induced hepatic steatosis. (A) GSH. (B) T-AOC. (C) SOD. (D) MDA. SiRNA PASK HepG2 cells and HepG2 cells incubated with increased 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 < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001. (Control, siRNA scrambled; NF, Nuciferine.)
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Figure 7: The effect of nuciferine or siRNA PASK on cell lipid peroxidation in HepG2 cells of OA-induced hepatic steatosis. (A) GSH. (B) T-AOC. (C) SOD. (D) MDA. SiRNA PASK HepG2 cells and HepG2 cells incubated with increased 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 < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001. (Control, siRNA scrambled; NF, Nuciferine.)

Mentions: To see whether nuciferine or siRNA PASK could ameliorate lipid peroxidation and oxidative stress, the main factors contributed to NASH, we tested the content of GSH, T-AOC and SOD, which were selected as measurement of lipid peroxidation. The GSH level in OA group was significantly depleted compared with the control (∗∗P < 0.01). Upon addition of nuciferine, the content of GSH was surprising decreased by 13.51, 46.66 and 49% (∗P < 0.05 vs. OA), and siRNA PASK depleted by 69.97% (∗P < 0.05 vs. OA), but vitamin E significantly increased GSH level by twofold compared with the OA group (∗∗∗P < 0.001 vs. OA) (Figure 7A). The T-AOC and SOD level in OA group was significantly depleted compared with the control (∗P < 0.05 and ∗∗P < 0.01), and addition of nuciferine could simultaneously reverse the decrease level of T-AOC (∗∗∗P < 0.001 vs. OA) and SOD (∗P < 0.05 vs. OA), while the siRNA PASK (∗∗∗P < 0.001 and ∗P < 0.05 vs. OA) and vitamin E (∗∗∗P < 0.001 vs. OA) showed similar trend (Figures 7B,C). In addition, MDA was regarded as measurement of oxidative stress. The OA treated group could significantly increase MDA content compared with the control (∗∗∗P < 0.001). Addition of nuciferine diminished MDA level by 33.57, 34.07 and 34.12 (∗∗P < 0.01 vs. OA), and vitamin E (25 μM) and siRNA PASK were inhibited by 27.62% (∗∗P < 0.01 vs. OA) and 35% (∗∗P < 0.01 vs. OA) compared with the OA group (Figure 7D). These findings demonstrate that nuciferine and siRNA PASK could both successfully decrease the oxidative stress, probably owing to the down-regulation of antioxidant molecules SOD, T-AOC, but not GSH.


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 cell lipid peroxidation in HepG2 cells of OA-induced hepatic steatosis. (A) GSH. (B) T-AOC. (C) SOD. (D) MDA. SiRNA PASK HepG2 cells and HepG2 cells incubated with increased 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 < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001. (Control, siRNA scrambled; NF, Nuciferine.)
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Figure 7: The effect of nuciferine or siRNA PASK on cell lipid peroxidation in HepG2 cells of OA-induced hepatic steatosis. (A) GSH. (B) T-AOC. (C) SOD. (D) MDA. SiRNA PASK HepG2 cells and HepG2 cells incubated with increased 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 < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001. (Control, siRNA scrambled; NF, Nuciferine.)
Mentions: To see whether nuciferine or siRNA PASK could ameliorate lipid peroxidation and oxidative stress, the main factors contributed to NASH, we tested the content of GSH, T-AOC and SOD, which were selected as measurement of lipid peroxidation. The GSH level in OA group was significantly depleted compared with the control (∗∗P < 0.01). Upon addition of nuciferine, the content of GSH was surprising decreased by 13.51, 46.66 and 49% (∗P < 0.05 vs. OA), and siRNA PASK depleted by 69.97% (∗P < 0.05 vs. OA), but vitamin E significantly increased GSH level by twofold compared with the OA group (∗∗∗P < 0.001 vs. OA) (Figure 7A). The T-AOC and SOD level in OA group was significantly depleted compared with the control (∗P < 0.05 and ∗∗P < 0.01), and addition of nuciferine could simultaneously reverse the decrease level of T-AOC (∗∗∗P < 0.001 vs. OA) and SOD (∗P < 0.05 vs. OA), while the siRNA PASK (∗∗∗P < 0.001 and ∗P < 0.05 vs. OA) and vitamin E (∗∗∗P < 0.001 vs. OA) showed similar trend (Figures 7B,C). In addition, MDA was regarded as measurement of oxidative stress. The OA treated group could significantly increase MDA content compared with the control (∗∗∗P < 0.001). Addition of nuciferine diminished MDA level by 33.57, 34.07 and 34.12 (∗∗P < 0.01 vs. OA), and vitamin E (25 μM) and siRNA PASK were inhibited by 27.62% (∗∗P < 0.01 vs. OA) and 35% (∗∗P < 0.01 vs. OA) compared with the OA group (Figure 7D). These findings demonstrate that nuciferine and siRNA PASK could both successfully decrease the oxidative stress, probably owing to the down-regulation of antioxidant molecules SOD, T-AOC, but not GSH.

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).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.

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