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Article Open Access

Peroxisome proliferator‑activated receptor α regulates acesulfame‑K‑induced NAFLD via hepatic PLCβ: Foe and friend

  • Authors:
    • Peng-Yao Lin
    • Jia-Rong Xie
    • Tian-Chen Qian
    • Shi-Song Wang
    • Si-Yi Yu
    • Wen-Bo Shi
    • Ying Wang
    • Lu-Ze Cen
    • Qing-Jing Zhu
    • Yi-Yang Zheng
    • Hui Gao
    • Rong Fang
    • Zhao-Xia Xia
    • Ai-Ming Liu
    • Lei Xu
  • View Affiliations / Copyright

    Affiliations: Department of Gastroenterology, The First Affiliated Hospital of Ningbo University, Ningbo, Zhejiang 315010, P.R. China, Department of Gastroenterology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310027, P.R. China, Department of Pharmacology, Health Science Center, Ningbo University, Ningbo, Zhejiang 315211, P.R. China, Ningbo Clinical Pathology Diagnosis Center, Ningbo, Zhejiang 315021, P.R. China
    Copyright: © Lin et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 102
    |
    Published online on: February 25, 2026
       https://doi.org/10.3892/ijmm.2026.5773
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Abstract

Food additive acesulfame‑K (AK), a non‑nutritive sweetener, is widely used as a low‑calorie sugar substitute to reduce energy intake. However, its potential impact on nonalcoholic fatty liver disease (NAFLD) and the involvement of peroxisome proliferator‑activated receptor α (PPARα) remain unclear. In the present study, male wild‑type (WT) and PPARα‑null (KO) mice fed a 60% high‑fat diet were treated with AK (2 mg/ml) in drinking water for 12 weeks to evaluate the effects of chronic AK exposure on NAFLD progression and the role of PPARα. PPARα inhibition and activation strategies were further applied in in vivo and in vitro models to validate the key findings. AK supplementation markedly increased hepatic lipid accumulation and impaired glucose tolerance through activation of phospholipase C beta (PLCβ) in hepatic sweet taste receptor (STR) signaling in the WT mice, but not in the KO mice. Consistently, PLCβ activation was observed in AK‑induced lipid accumulation in Hepa1‑6 and Huh‑7 cells and was abolished by PPARα knockdown or inhibition. Pharmacological activation of PPARα mitigated AK‑induced NAFLD progression by suppressing PLCβ activation in STR signaling. These findings demonstrated that chronic AK intake exacerbates NAFLD progression via PLCβ activation in hepatic STR signaling and that PLCβ activation depends on physiological PPARα activity. Pharmacological PPARα activation exerts a protective effect, highlighting the dual roles of PPARα in regulating AK‑associated NAFLD risk.
View Figures

Figure 1

AK intake does not modify the
phenotype of the WT or KO mice. (A) Body weight gain of the WT and
KO mice. (B) Cumulative food intake of the WT and KO mice. (C)
Ratio of total food intake to body weight per 10 g in the WT and KO
mice. (D) Cumulative water intake of the WT and KO mice. (E) Ratio
of total water intake to body weight per 10 g in the WT and KO
mice. (F) Liver indices of the WT and KO mice. The data were
expressed as the means ± SEM. *P<0.05 and
***P<0.0001 WT vs. KO mice; #P<0.05
WT-AK vs. KO-AK mice; ns. not significant. Differences at the time
points were determined by two-way ANOVA and Bonferroni post hoc
correction. AK, acesulfame-K; BW, body weight; KO, PPARα-null; WT,
wild-type; PPARα, peroxisome proliferator-activated receptor α.

Figure 2

AK promotes NAFLD development in the
WT mice not the KO mice. (A) Hematoxylin and eosin staining of
liver tissues from each group (magnification, ×40 and ×100). (B)
Liver TG and (C) TC concentrations in each group. (D) Serum TG and
(E) TC concentrations in each group. The data were expressed as the
means ± SEM. **P<0.01, ***P<0.0001, ns.
not significant. AK, acesulfame-K; NAFLD, nonalcoholic fatty liver
disease; WT, wild-type; KO, PPARα-null; TC, total cholesterol; TG,
triglycerides; PPARα, peroxisome proliferator-activated receptor
α.

Figure 3

AK impairs glucose homeostasis via
activating hepatic PLCβ in STR signaling. (A) HFD-fed WT and KO
mice receiving normal water or AK supplementation (12-week
intervention, n=5). Metabolic tests (OGTT/ITT) at weeks 11-12. (B)
OGTT and (C) the corresponding AUCs in each group. (D) Fasting
blood glucose levels in the different groups of mice after 6 h of
fasting. (E) ITT and (F) the corresponding AUCs in the WT and WT-AK
mice. The average changes in (G) Gck, (H) G6pc1, (I)
Slc2a2 and (J) Slc5a10 transcription in the 4 groups.
(K) PAS staining of liver tissues from each group (magnification,
×100). (L) Liver glycogen content in each group. (M) Liver protein
expression levels of PLCβ, p-PLCβ, TAS1R2, and TAS1R3 were measured
using western blot analysis. The data were expressed as the means ±
SEM. *P<0.05, **P<0.01,
***P<0.0001, ns. not significant. AK, acesulfame-K;
p-, phosphorylated; PLCβ, phospholipase C beta; STR, sweet taste
receptor; HFD, high-fat diet; WT, wild-type; KO, PPARα-null; OGTT,
oral glucose tolerance test; ITT, insulin tolerance test; AUC, area
under the curve; G6pc1, glucose-6-phosphatase catalytic subunit
1; Gck, glucokinase; Slc2a2, solute carrier family 2,
member 2; Slc5a10, solute carrier family 5, member 10; TAS1R,
taste 1 receptor member.

Figure 4

PLCβ is activated in lipid
accumulation increased by AK in Hepa1-6 and Huh-7 cells. (A) Oil
Red O staining of Hepa1-6 and Huh-7 cells after 48 h of treatment
with FFA combined with AK. (B) TG content in the cytoplasm of
Hepa1-6 cells. (C) TG content in the cytoplasm of Huh-7 cells. (D)
PLCβ phosphorylation in Hepa1-6 cells during a time course of AK
treatment. (E) PLCβ phosphorylation in Hepa1-6 cells after 1 h of
AK treatment. The data were expressed as the means ± SEM.
**P<0.01, ***P<0.0001. PLCβ,
phospholipase C beta; AK, acesulfame-K; Hepa1-6, mouse hepatoma
cells; Huh-7, human hepatocellular carcinoma cells; FFA, free fatty
acid; TG, triglycerides; WT, wild-type; NC, control group without
free fatty acid treatment; CON, control group without added
acesulfame-K but with free fatty acids added; KO, PPARα-null; p-,
phosphorylated.

Figure 5

PLCβ activation and lipid
accumulation induced by AK are deleted by PPARα downregulation. (A)
Pparα mRNA levels in Hepa1-6 cells transfected with a vector
plasmid and three shPparα plasmids. (B) PPARα protein levels
in Hepa1-6 cells transfected with a vector plasmid and three
shPparα plasmids. (C) PLCβ phosphorylation in Hepa1-6 cells
transfected with shPparα-2 after 1 h of AK treatment. (D)
Oil Red O staining of Hepa1-6 cells after transfection with the
shPparα-2 plasmid. (E) TG content in the cytoplasm of
Hepa1-6 cells transfected with the shPparα-2 plasmid. (F)
Oil Red O staining of Hepa1-6 cells treated with GW6,471. (G) TG
concentration in Hepa1-6 cells treated with GW6,471. The data were
expressed as the means ± SEM. **P<0.01,
***P<0.001, ****P<0.0001. AK, acesulfame-K; PLCβ,
phospholipase C beta; PPARα, peroxisome proliferator-activated
receptor α; sh, short hairpin; Hepa1-6, mouse hepatoma cells; TG,
triglycerides; CON, control group without added acesulfame-K but
with free fatty acids added; FFA, free fatty acid; GW6,471, a
peroxisome proliferator-activated receptor α inhibitor; KO,
PPARα-null; NC, control group without free fatty acid treatment;
p-, phosphorylated; shNC, short hairpin RNA negative control; TC,
total cholesterol; WT, wild-type.

Figure 6

Multiple targets including PLCβ in
STR signaling are regulated by PPARα. Upregulation of PPARα target
genes (A) Acot1 and (C) Ehhadh by the agonists
fenofibrate and Wy-14,643. (B and D) Pharmacological regulation of
TAS1R2, TAS1R3, GNAT3, and PLCβ induced by fenofibrate and
Wy-14,643. (E) Volcano plot depicting hepatic transcriptomic
profiles of Wy14,643-treated mice vs. vehicle controls. (F) Heatmap
of diacylglycerol kinase family genes revealed Dgkh as a
potential PPARα target gene. (G) DAG levels in the liver of the WT,
WT-AK mice were lower than those in the KO and KO-AK mice. (H)
Dgkh transcription regulated by fenofibrate and Wy-14,643
detected using quantitative PCR. (I) Validation of DGKH expression
regulated by PPARα agonists fenofibrate and Wy-14,643. (J) Hepatic
DAG content in experimental groups. The data were expressed as the
means ± SEM. *P<0.05, **P<0.01,
***P<0.001, and ****P<0.0001. PLCβ,
phospholipase C beta; STR, sweet taste receptor; PPARα, peroxisome
proliferator-activated receptor PPARα, peroxisome proliferator-act;
Acot, acyl-CoA thioesterase; Ehhadh, enoyl-CoA
hydratase/3-hydroxyacyl-CoA dehydrogenase; TAS1R, taste 1
receptor member; GNAT3, G protein subunit alpha transducin 3; Wy,
Wy-14,643, a peroxisome proliferator-activated receptor alpha
agonist; Dgkh, diacylglycerol kinase eta; KO, PPARα-null;
DGKH, diacylglycerol kinase η; DAG, diacylglycerol; WT, wild-type;
CON, control group; FC, fold change; PLCβ, phospholipase C
beta.

Figure 7

PLCβ is inhibited in steatosis
attenuated by pharmacologically-activated PPARα. (A) HFD-fed WT and
KO mice receiving normal water or AK supplementation (12-week
intervention, n=3). During weeks 16-18, control solvent or F (25
mg/kg) was administered twice daily via oral gavage. (B)
Hematoxylin and eosin staining of liver tissues from each group
(magnification, ×100). (C) Liver TG concentrations in each group.
(D) The PLCβ protein expression and phosphorylation status were
quantified in each group. Data were expressed as the means ± SEM.
*P<0.05, ***P<0.0001, ns. not
significant. PLCβ, phospholipase C beta; PPARα, peroxisome
proliferator-activated receptor α; HFD, high-fat diet; WT,
wild-type; KO, PPARα-null; AK, acesulfame-K; TG, triglycerides; F,
fenofibrate; p-, phosphorylated.

Figure 8

Proposed schematic mechanism by which
AK promotes the development of NAFLD dependent on PPARα. The
physiological actions of PPARα involve adaptive regulation of STRs,
activation of PLCβ, and transcription of DGKH, which mediates DAG
catabolism. Pharmacological actions of PPARα downregulate STR
signaling via modulation of GNAT3 and PLCβ expression. AK,
acesulfame-K; NAFLD, nonalcoholic fatty liver disease; PPARα,
peroxisome proliferator-activated receptor α; STR, sweet taste
receptor; PLCβ, phospholipase C beta; DGKH, diacylglycerol kinase
eta; DAG, diacylglycerol; GNAT3, G protein subunit alpha transducin
3; TAS1R, taste 1 receptor member; PA, phosphatidic acid.
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Copy and paste a formatted citation
Spandidos Publications style
Lin P, Xie J, Qian T, Wang S, Yu S, Shi W, Wang Y, Cen L, Zhu Q, Zheng Y, Zheng Y, et al: Peroxisome proliferator‑activated receptor &alpha; regulates acesulfame‑K‑induced NAFLD via hepatic PLC&beta;: Foe and friend. Int J Mol Med 57: 102, 2026.
APA
Lin, P., Xie, J., Qian, T., Wang, S., Yu, S., Shi, W. ... Xu, L. (2026). Peroxisome proliferator‑activated receptor &alpha; regulates acesulfame‑K‑induced NAFLD via hepatic PLC&beta;: Foe and friend. International Journal of Molecular Medicine, 57, 102. https://doi.org/10.3892/ijmm.2026.5773
MLA
Lin, P., Xie, J., Qian, T., Wang, S., Yu, S., Shi, W., Wang, Y., Cen, L., Zhu, Q., Zheng, Y., Gao, H., Fang, R., Xia, Z., Liu, A., Xu, L."Peroxisome proliferator‑activated receptor &alpha; regulates acesulfame‑K‑induced NAFLD via hepatic PLC&beta;: Foe and friend". International Journal of Molecular Medicine 57.4 (2026): 102.
Chicago
Lin, P., Xie, J., Qian, T., Wang, S., Yu, S., Shi, W., Wang, Y., Cen, L., Zhu, Q., Zheng, Y., Gao, H., Fang, R., Xia, Z., Liu, A., Xu, L."Peroxisome proliferator‑activated receptor &alpha; regulates acesulfame‑K‑induced NAFLD via hepatic PLC&beta;: Foe and friend". International Journal of Molecular Medicine 57, no. 4 (2026): 102. https://doi.org/10.3892/ijmm.2026.5773
Copy and paste a formatted citation
x
Spandidos Publications style
Lin P, Xie J, Qian T, Wang S, Yu S, Shi W, Wang Y, Cen L, Zhu Q, Zheng Y, Zheng Y, et al: Peroxisome proliferator‑activated receptor &alpha; regulates acesulfame‑K‑induced NAFLD via hepatic PLC&beta;: Foe and friend. Int J Mol Med 57: 102, 2026.
APA
Lin, P., Xie, J., Qian, T., Wang, S., Yu, S., Shi, W. ... Xu, L. (2026). Peroxisome proliferator‑activated receptor &alpha; regulates acesulfame‑K‑induced NAFLD via hepatic PLC&beta;: Foe and friend. International Journal of Molecular Medicine, 57, 102. https://doi.org/10.3892/ijmm.2026.5773
MLA
Lin, P., Xie, J., Qian, T., Wang, S., Yu, S., Shi, W., Wang, Y., Cen, L., Zhu, Q., Zheng, Y., Gao, H., Fang, R., Xia, Z., Liu, A., Xu, L."Peroxisome proliferator‑activated receptor &alpha; regulates acesulfame‑K‑induced NAFLD via hepatic PLC&beta;: Foe and friend". International Journal of Molecular Medicine 57.4 (2026): 102.
Chicago
Lin, P., Xie, J., Qian, T., Wang, S., Yu, S., Shi, W., Wang, Y., Cen, L., Zhu, Q., Zheng, Y., Gao, H., Fang, R., Xia, Z., Liu, A., Xu, L."Peroxisome proliferator‑activated receptor &alpha; regulates acesulfame‑K‑induced NAFLD via hepatic PLC&beta;: Foe and friend". International Journal of Molecular Medicine 57, no. 4 (2026): 102. https://doi.org/10.3892/ijmm.2026.5773
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