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Involvement of KLF11 in hepatic glucose metabolism in mice via suppressing of PEPCK-C expression.

Zhang H, Chen Q, Jiao T, Cui A, Sun X, Fang W, Xie L, Liu Y, Fang F, Chang Y - PLoS ONE (2014)

Bottom Line: Abnormal hepatic gluconeogenesis is related to hyperglycemia in mammals with insulin resistance.Despite the strong evidences linking Krüppel-like factor 11 (KLF11) gene mutations to development of Type 2 diabetes, the precise physiological functions of KLF11 in vivo remain largely unknown.Our data strongly indicated the involvement of KLF11 in hepatic glucose homeostasis via modulating the expression of PEPCK-C.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry & Molecular Biology, School of Basic Medicine, Anhui Medical University, Hefei, China ; National Laboratory of Medical Molecular Biology, Institute of Basic Medical Science, Chinese Academy of Medical Science and Peking Union Medical College, Beijing, China.

ABSTRACT

Background: Abnormal hepatic gluconeogenesis is related to hyperglycemia in mammals with insulin resistance. Despite the strong evidences linking Krüppel-like factor 11 (KLF11) gene mutations to development of Type 2 diabetes, the precise physiological functions of KLF11 in vivo remain largely unknown.

Results: In current investigation, we showed that KLF11 is involved in modulating hepatic glucose metabolism in mice. Overexpression of KLF11 in primary mouse hepatocytes could inhibit the expression of gluconeogenic genes, including phosphoenolpyruvate carboxykinase (cytosolic isoform, PEPCK-C) and peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), subsequently decreasing the cellular glucose output. Diabetic mice with overexpression of KLF11 gene in livers significantly ameliorated hyperglycemia and glucose intolerance; in contrast, the knockdown of KLF11 expression in db/m and C57BL/6J mice livers impaired glucose tolerance.

Conclusions: Our data strongly indicated the involvement of KLF11 in hepatic glucose homeostasis via modulating the expression of PEPCK-C.

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Related in: MedlinePlus

KLF11 regulates the expression of the gluconeogenic genes in HepG2 cells and mouse primary hepatocytes.(A) qRT-PCR analysis of the mRNA expression levels of gluconeogenic genes in HepG2 cells infected with adenoviruses Ad-GFP or Ad-KLF11. (B) Western blot analysis of the protein levels of PEPCK-C in HepG2 cells infected with Ad-GFP or Ad-KLF11 adenoviruses. GADPH was used to show the similar amount of protein loaded in different lanes (Left panel). The relative intensities of PEPCK-C bands on the Western blot were determined using NIH Image 1.62 software and normalized using GADPH band intensity (Right panel). (C) qRT-PCR analysis of the mRNA expression levels of gluconeogenic genes in primary hepatocytes infected with adenoviruses Ad-GFP or Ad-KLF11. At 24 hr after infection, hepatocytes were switched to starvation media for 6 hr, followed by treatment with 10 µM forskolin and 1 µM dexamethasone for 1.5 hr. (D) Measurement of cellular glucose production in primary hepatocytes as described in Fig. 3C. (E) qRT-PCR analysis showing the expression levels of gluconeogenic genes in primary hepatocytes infected with control Ad-shCon or Ad-shKLF11. Hepatocytes were grown for 2 days post-infection in RPMI-1640+10% FBS. The data shown were the means ± SEM (n = 3). Statistical significance was determined using a two-tailed Student’s t-test. * on each bar indicated the comparison between Ad-KFL11 and Ad-GFP infection (*P<0.05, **P<0.01).
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pone-0089552-g003: KLF11 regulates the expression of the gluconeogenic genes in HepG2 cells and mouse primary hepatocytes.(A) qRT-PCR analysis of the mRNA expression levels of gluconeogenic genes in HepG2 cells infected with adenoviruses Ad-GFP or Ad-KLF11. (B) Western blot analysis of the protein levels of PEPCK-C in HepG2 cells infected with Ad-GFP or Ad-KLF11 adenoviruses. GADPH was used to show the similar amount of protein loaded in different lanes (Left panel). The relative intensities of PEPCK-C bands on the Western blot were determined using NIH Image 1.62 software and normalized using GADPH band intensity (Right panel). (C) qRT-PCR analysis of the mRNA expression levels of gluconeogenic genes in primary hepatocytes infected with adenoviruses Ad-GFP or Ad-KLF11. At 24 hr after infection, hepatocytes were switched to starvation media for 6 hr, followed by treatment with 10 µM forskolin and 1 µM dexamethasone for 1.5 hr. (D) Measurement of cellular glucose production in primary hepatocytes as described in Fig. 3C. (E) qRT-PCR analysis showing the expression levels of gluconeogenic genes in primary hepatocytes infected with control Ad-shCon or Ad-shKLF11. Hepatocytes were grown for 2 days post-infection in RPMI-1640+10% FBS. The data shown were the means ± SEM (n = 3). Statistical significance was determined using a two-tailed Student’s t-test. * on each bar indicated the comparison between Ad-KFL11 and Ad-GFP infection (*P<0.05, **P<0.01).

Mentions: KLF11 inhibited the PEPCK-C promoter activity and mutation of the GC-rich sequence in promoter sequence abolished KLF11-mediated suppression. Subsequently, we sought to pursue the functional importance of KLF11 in gluconeogenic program in vitro. HepG2 cells were infected with adenovirus expressing KLF11 or GFP (control). Overexpression of KLF11 decreased PEPCK-C expression level for both mRNA and protein (∼70%) in HepG2 cells (Fig. 3A), whereas the expression levels of PGC-1α and G6pase were not markedly affected (Fig. 3A and B). A similar experiment was performed in primary hepatocytes, although overexpression of KLF11 by Ad-KLF11 did not affect the expression of gluconeogenic genes in the primary hepatocytes under basal conditions (data not shown), it significantly inhibited the expression of these genes (including PGC-1α, PEPCK-C, and G6pase) in the presence of forskolin (Fsk) and dexamethasone (Dex), which mimic the fasting action of glucagons and glucocorticoids, respectively (Fig. 3C). The ability of KLF11 to inhibit the gluconeogenic program in hepatocytes suggested that it may decrease cellular glucose output. As expected, Ad-KLF11 infection reduced glucose production in primary mouse hepatocytes in the presence of FSK and Dex (Fig. 3D). In contrast, Ad-shKLF11-mediated reduction of KLF11 expression in primary hepatocytes modestly increased the expression levels of PEPCK-C and G6pase (Fig. 3E).


Involvement of KLF11 in hepatic glucose metabolism in mice via suppressing of PEPCK-C expression.

Zhang H, Chen Q, Jiao T, Cui A, Sun X, Fang W, Xie L, Liu Y, Fang F, Chang Y - PLoS ONE (2014)

KLF11 regulates the expression of the gluconeogenic genes in HepG2 cells and mouse primary hepatocytes.(A) qRT-PCR analysis of the mRNA expression levels of gluconeogenic genes in HepG2 cells infected with adenoviruses Ad-GFP or Ad-KLF11. (B) Western blot analysis of the protein levels of PEPCK-C in HepG2 cells infected with Ad-GFP or Ad-KLF11 adenoviruses. GADPH was used to show the similar amount of protein loaded in different lanes (Left panel). The relative intensities of PEPCK-C bands on the Western blot were determined using NIH Image 1.62 software and normalized using GADPH band intensity (Right panel). (C) qRT-PCR analysis of the mRNA expression levels of gluconeogenic genes in primary hepatocytes infected with adenoviruses Ad-GFP or Ad-KLF11. At 24 hr after infection, hepatocytes were switched to starvation media for 6 hr, followed by treatment with 10 µM forskolin and 1 µM dexamethasone for 1.5 hr. (D) Measurement of cellular glucose production in primary hepatocytes as described in Fig. 3C. (E) qRT-PCR analysis showing the expression levels of gluconeogenic genes in primary hepatocytes infected with control Ad-shCon or Ad-shKLF11. Hepatocytes were grown for 2 days post-infection in RPMI-1640+10% FBS. The data shown were the means ± SEM (n = 3). Statistical significance was determined using a two-tailed Student’s t-test. * on each bar indicated the comparison between Ad-KFL11 and Ad-GFP infection (*P<0.05, **P<0.01).
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pone-0089552-g003: KLF11 regulates the expression of the gluconeogenic genes in HepG2 cells and mouse primary hepatocytes.(A) qRT-PCR analysis of the mRNA expression levels of gluconeogenic genes in HepG2 cells infected with adenoviruses Ad-GFP or Ad-KLF11. (B) Western blot analysis of the protein levels of PEPCK-C in HepG2 cells infected with Ad-GFP or Ad-KLF11 adenoviruses. GADPH was used to show the similar amount of protein loaded in different lanes (Left panel). The relative intensities of PEPCK-C bands on the Western blot were determined using NIH Image 1.62 software and normalized using GADPH band intensity (Right panel). (C) qRT-PCR analysis of the mRNA expression levels of gluconeogenic genes in primary hepatocytes infected with adenoviruses Ad-GFP or Ad-KLF11. At 24 hr after infection, hepatocytes were switched to starvation media for 6 hr, followed by treatment with 10 µM forskolin and 1 µM dexamethasone for 1.5 hr. (D) Measurement of cellular glucose production in primary hepatocytes as described in Fig. 3C. (E) qRT-PCR analysis showing the expression levels of gluconeogenic genes in primary hepatocytes infected with control Ad-shCon or Ad-shKLF11. Hepatocytes were grown for 2 days post-infection in RPMI-1640+10% FBS. The data shown were the means ± SEM (n = 3). Statistical significance was determined using a two-tailed Student’s t-test. * on each bar indicated the comparison between Ad-KFL11 and Ad-GFP infection (*P<0.05, **P<0.01).
Mentions: KLF11 inhibited the PEPCK-C promoter activity and mutation of the GC-rich sequence in promoter sequence abolished KLF11-mediated suppression. Subsequently, we sought to pursue the functional importance of KLF11 in gluconeogenic program in vitro. HepG2 cells were infected with adenovirus expressing KLF11 or GFP (control). Overexpression of KLF11 decreased PEPCK-C expression level for both mRNA and protein (∼70%) in HepG2 cells (Fig. 3A), whereas the expression levels of PGC-1α and G6pase were not markedly affected (Fig. 3A and B). A similar experiment was performed in primary hepatocytes, although overexpression of KLF11 by Ad-KLF11 did not affect the expression of gluconeogenic genes in the primary hepatocytes under basal conditions (data not shown), it significantly inhibited the expression of these genes (including PGC-1α, PEPCK-C, and G6pase) in the presence of forskolin (Fsk) and dexamethasone (Dex), which mimic the fasting action of glucagons and glucocorticoids, respectively (Fig. 3C). The ability of KLF11 to inhibit the gluconeogenic program in hepatocytes suggested that it may decrease cellular glucose output. As expected, Ad-KLF11 infection reduced glucose production in primary mouse hepatocytes in the presence of FSK and Dex (Fig. 3D). In contrast, Ad-shKLF11-mediated reduction of KLF11 expression in primary hepatocytes modestly increased the expression levels of PEPCK-C and G6pase (Fig. 3E).

Bottom Line: Abnormal hepatic gluconeogenesis is related to hyperglycemia in mammals with insulin resistance.Despite the strong evidences linking Krüppel-like factor 11 (KLF11) gene mutations to development of Type 2 diabetes, the precise physiological functions of KLF11 in vivo remain largely unknown.Our data strongly indicated the involvement of KLF11 in hepatic glucose homeostasis via modulating the expression of PEPCK-C.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry & Molecular Biology, School of Basic Medicine, Anhui Medical University, Hefei, China ; National Laboratory of Medical Molecular Biology, Institute of Basic Medical Science, Chinese Academy of Medical Science and Peking Union Medical College, Beijing, China.

ABSTRACT

Background: Abnormal hepatic gluconeogenesis is related to hyperglycemia in mammals with insulin resistance. Despite the strong evidences linking Krüppel-like factor 11 (KLF11) gene mutations to development of Type 2 diabetes, the precise physiological functions of KLF11 in vivo remain largely unknown.

Results: In current investigation, we showed that KLF11 is involved in modulating hepatic glucose metabolism in mice. Overexpression of KLF11 in primary mouse hepatocytes could inhibit the expression of gluconeogenic genes, including phosphoenolpyruvate carboxykinase (cytosolic isoform, PEPCK-C) and peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), subsequently decreasing the cellular glucose output. Diabetic mice with overexpression of KLF11 gene in livers significantly ameliorated hyperglycemia and glucose intolerance; in contrast, the knockdown of KLF11 expression in db/m and C57BL/6J mice livers impaired glucose tolerance.

Conclusions: Our data strongly indicated the involvement of KLF11 in hepatic glucose homeostasis via modulating the expression of PEPCK-C.

Show MeSH
Related in: MedlinePlus