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Role of AMP-Activated Protein Kinase (AMPK) in Smoking-Induced Lung Inflammation and Emphysema.

Lee JS, Park SJ, Cho YS, Huh JW, Oh YM, Lee SD - Tuberc Respir Dis (Seoul) (2015)

Bottom Line: Furthermore, there is recent evidence that it participates in limiting acute inflammatory reactions, apoptosis and cellular senescence.We then investigated the role of AMPK in the induction of interleukin-8 (IL-8) by cigarette smoke extract (CSE) in A549 cells.CSE increased AMPK activation in a CSE concentration- and time-dependent manner. 5-Aminoimidazole-4-carboxamide-1-β-4-ribofuranoside (AICAR), an AMPK activator, decreased CSE-induced IL-8 production while Compound C, an AMPK inhibitor, increased it, as did pretreatment with an AMPKα1-specific small interfering RNA.

View Article: PubMed Central - PubMed

Affiliation: Department of Pulmonary and Critical Care Medicine and Clinical Research Center for Chronic Obstructive Airway Diseases, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea.

ABSTRACT

Background: AMP-activated protein kinase (AMPK) not only functions as an intracellular energy sensor and regulator, but is also a general sensor of oxidative stress. Furthermore, there is recent evidence that it participates in limiting acute inflammatory reactions, apoptosis and cellular senescence. Thus, it may oppose the development of chronic obstructive pulmonary disease.

Methods: To investigate the role of AMPK in cigarette smoke-induced lung inflammation and emphysema we first compared cigarette smoking and polyinosinic-polycytidylic acid [poly(I:C)]-induced lung inflammation and emphysema in AMPKα1-deficient (AMPKα1-HT) mice and wild-type mice of the same genetic background. We then investigated the role of AMPK in the induction of interleukin-8 (IL-8) by cigarette smoke extract (CSE) in A549 cells.

Results: Cigarette smoking and poly(I:C)-induced lung inflammation and emphysema were elevated in AMPKα1-HT compared to wild-type mice. CSE increased AMPK activation in a CSE concentration- and time-dependent manner. 5-Aminoimidazole-4-carboxamide-1-β-4-ribofuranoside (AICAR), an AMPK activator, decreased CSE-induced IL-8 production while Compound C, an AMPK inhibitor, increased it, as did pretreatment with an AMPKα1-specific small interfering RNA.

Conclusion: AMPKα1-deficient mice have increased susceptibility to lung inflammation and emphysema when exposed to cigarette smoke, and AMPK appears to reduce lung inflammation and emphysema by lowering IL-8 production.

No MeSH data available.


Related in: MedlinePlus

Generation of heterozygous AMPKα1-deficient (AMPKα1-HT) C57BL/6 mice. (A) Genotyping of mice by multiplex polymerase chain reaction; this yielded amplification products of 350 bp (knockout) and 450 bp (wild-type). (B) Western blot analysis of AMPKα expression in lung tissue. AMPK: AMP-activated protein kinase; WT: wild-type littermate; HT: heterozygous AMPKα1-deficient mouse; M: marker. Data represent mean±SEM (n=3). Each data point is based on three independent Western blots. *p<0.05, vs. WT mice.
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Figure 1: Generation of heterozygous AMPKα1-deficient (AMPKα1-HT) C57BL/6 mice. (A) Genotyping of mice by multiplex polymerase chain reaction; this yielded amplification products of 350 bp (knockout) and 450 bp (wild-type). (B) Western blot analysis of AMPKα expression in lung tissue. AMPK: AMP-activated protein kinase; WT: wild-type littermate; HT: heterozygous AMPKα1-deficient mouse; M: marker. Data represent mean±SEM (n=3). Each data point is based on three independent Western blots. *p<0.05, vs. WT mice.

Mentions: Routine genotyping was carried out by multiplex polymerase chain reaction with primers synthesized by Bioneer (Daejeon, Korea). Reaction conditions were as follows: 5 minutes at 94℃, 35 cycles of 94℃ for 30 seconds, 60℃ for 30 seconds and 72℃ for 1 minute, and 5 minutes at 72℃. Primers were as follows: for AMPKα1 knockout, forward (F) 5'-GGGCTGCAGBAATTCGATAT CAAGC-3' and reverse (R) 5'-CCTTCCTGAAATBACTTCTG GTGC-3', and for AMPK wild type, forward (F) 5'-AGCCGACT TTGGTAAGGATG-3', and reverse (R) 5'-CCCACTTTCCATTT TCTCCA-3', yielding amplification products of 350 bp and 450 bp, respectively (Figure 1A).


Role of AMP-Activated Protein Kinase (AMPK) in Smoking-Induced Lung Inflammation and Emphysema.

Lee JS, Park SJ, Cho YS, Huh JW, Oh YM, Lee SD - Tuberc Respir Dis (Seoul) (2015)

Generation of heterozygous AMPKα1-deficient (AMPKα1-HT) C57BL/6 mice. (A) Genotyping of mice by multiplex polymerase chain reaction; this yielded amplification products of 350 bp (knockout) and 450 bp (wild-type). (B) Western blot analysis of AMPKα expression in lung tissue. AMPK: AMP-activated protein kinase; WT: wild-type littermate; HT: heterozygous AMPKα1-deficient mouse; M: marker. Data represent mean±SEM (n=3). Each data point is based on three independent Western blots. *p<0.05, vs. WT mice.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4311035&req=5

Figure 1: Generation of heterozygous AMPKα1-deficient (AMPKα1-HT) C57BL/6 mice. (A) Genotyping of mice by multiplex polymerase chain reaction; this yielded amplification products of 350 bp (knockout) and 450 bp (wild-type). (B) Western blot analysis of AMPKα expression in lung tissue. AMPK: AMP-activated protein kinase; WT: wild-type littermate; HT: heterozygous AMPKα1-deficient mouse; M: marker. Data represent mean±SEM (n=3). Each data point is based on three independent Western blots. *p<0.05, vs. WT mice.
Mentions: Routine genotyping was carried out by multiplex polymerase chain reaction with primers synthesized by Bioneer (Daejeon, Korea). Reaction conditions were as follows: 5 minutes at 94℃, 35 cycles of 94℃ for 30 seconds, 60℃ for 30 seconds and 72℃ for 1 minute, and 5 minutes at 72℃. Primers were as follows: for AMPKα1 knockout, forward (F) 5'-GGGCTGCAGBAATTCGATAT CAAGC-3' and reverse (R) 5'-CCTTCCTGAAATBACTTCTG GTGC-3', and for AMPK wild type, forward (F) 5'-AGCCGACT TTGGTAAGGATG-3', and reverse (R) 5'-CCCACTTTCCATTT TCTCCA-3', yielding amplification products of 350 bp and 450 bp, respectively (Figure 1A).

Bottom Line: Furthermore, there is recent evidence that it participates in limiting acute inflammatory reactions, apoptosis and cellular senescence.We then investigated the role of AMPK in the induction of interleukin-8 (IL-8) by cigarette smoke extract (CSE) in A549 cells.CSE increased AMPK activation in a CSE concentration- and time-dependent manner. 5-Aminoimidazole-4-carboxamide-1-β-4-ribofuranoside (AICAR), an AMPK activator, decreased CSE-induced IL-8 production while Compound C, an AMPK inhibitor, increased it, as did pretreatment with an AMPKα1-specific small interfering RNA.

View Article: PubMed Central - PubMed

Affiliation: Department of Pulmonary and Critical Care Medicine and Clinical Research Center for Chronic Obstructive Airway Diseases, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea.

ABSTRACT

Background: AMP-activated protein kinase (AMPK) not only functions as an intracellular energy sensor and regulator, but is also a general sensor of oxidative stress. Furthermore, there is recent evidence that it participates in limiting acute inflammatory reactions, apoptosis and cellular senescence. Thus, it may oppose the development of chronic obstructive pulmonary disease.

Methods: To investigate the role of AMPK in cigarette smoke-induced lung inflammation and emphysema we first compared cigarette smoking and polyinosinic-polycytidylic acid [poly(I:C)]-induced lung inflammation and emphysema in AMPKα1-deficient (AMPKα1-HT) mice and wild-type mice of the same genetic background. We then investigated the role of AMPK in the induction of interleukin-8 (IL-8) by cigarette smoke extract (CSE) in A549 cells.

Results: Cigarette smoking and poly(I:C)-induced lung inflammation and emphysema were elevated in AMPKα1-HT compared to wild-type mice. CSE increased AMPK activation in a CSE concentration- and time-dependent manner. 5-Aminoimidazole-4-carboxamide-1-β-4-ribofuranoside (AICAR), an AMPK activator, decreased CSE-induced IL-8 production while Compound C, an AMPK inhibitor, increased it, as did pretreatment with an AMPKα1-specific small interfering RNA.

Conclusion: AMPKα1-deficient mice have increased susceptibility to lung inflammation and emphysema when exposed to cigarette smoke, and AMPK appears to reduce lung inflammation and emphysema by lowering IL-8 production.

No MeSH data available.


Related in: MedlinePlus