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The inflammatory response seen when human omental adipose tissue explants are incubated in primary culture is not dependent upon albumin and is primarily in the nonfat cells.

Fain JN, Cheema P, Tichansky DS, Madan AK - J Inflamm (Lond) (2010)

Bottom Line: In the nonfat cells derived by a 2 h collagenase digestion of omental fat there was an inflammatory response comparable but not greater than that seen in tissue.The inflammatory response was not seen with respect to omentin/intelectin.The marked inflammatory response seen when human omental adipose tissue is incubated in vitro is reduced but not abolished in the presence of albumin with respect to IL-1beta, TNFalpha, IL-8, and is primarily in the nonfat cells of adipose tissue.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Molecular Sciences, College of Medicine, University of Tennessee Health Science Center, Memphis, TN 38163, USA.

ABSTRACT

Background: The present studies were designed to investigate the changes in gene expression during in vitro incubation of human visceral omental adipose tissue explants as well as fat cells and nonfat cells derived from omental fat.

Methods: Adipose tissue was obtained from extremely obese women undergoing bariatric surgery. Explants of the tissue as well as fat cells and the nonfat cells derived by digestion with collagenase were incubated for 20 minutes to 48 h. The expression of interleukin 1beta [IL-1beta], tumor necrosis factor alpha [TNFalpha], interleukin 8 [IL-8], NFkappaB(1)p50 subunit, hypoxia-inducible factor 1alpha [HIF1alpha], omentin/intelectin, and 11beta-hydroxysteroid dehydrogenase 1 [11beta-HSD1] mRNA were measured by qPCR as well as the release of IL-8 and TNFalpha.

Results: There was an inflammatory response at 2 h in explants of omental adipose tissue that was reduced but not abolished in the absence of albumin from the incubation buffer for IL-8, IL-1beta and TNFalpha. There was also an inflammatory response with regard to upregulation of HIF1alpha and NFkappaB1 gene expression that was unaffected whether albumin was present or absent from the medium. In the nonfat cells derived by a 2 h collagenase digestion of omental fat there was an inflammatory response comparable but not greater than that seen in tissue. The exception was HIF1alpha where the marked increase in gene expression was primarily seen in intact tissue. The inflammatory response was not seen with respect to omentin/intelectin. Over a subsequent 48 h incubation there was a marked increase in IL-8 mRNA expression and IL-8 release in adipose tissue explants that was also seen to the same extent in the nonfat cells incubated in the absence of fat cells.

Conclusion: The marked inflammatory response seen when human omental adipose tissue is incubated in vitro is reduced but not abolished in the presence of albumin with respect to IL-1beta, TNFalpha, IL-8, and is primarily in the nonfat cells of adipose tissue.

No MeSH data available.


Related in: MedlinePlus

Upregulation of the inflammatory response is rapid in onset. Explants of human omental adipose tissue were incubated for the indicated times and samples were taken from the medium to examine IL-8 release. The values in panel A are the means of two experiments. The values for IL-8 mRNA [log2 scale] in panel B are the means ± SEM of the ratios of mRNA at the indicated times to that at the start of the incubation for 5 experiments from as many different individuals. The values in panels B & C are based on the changes in absolute Cp values over time as compared to the Cp value in the unincubated tissue. Statistically significant changes in mRNA are indicated as follows: * P < 0.05 and ** P < 0.025. The IL-8 mRNA values at 1 h and all later times shown in panel B were statistically significant from the value at 0.33 h: P < 0.05. The values in panel C for IL-8, IL-1β, TNFα, NFκB1 and HIF-1α mRNA are from a different series of 4 experiments.
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Figure 1: Upregulation of the inflammatory response is rapid in onset. Explants of human omental adipose tissue were incubated for the indicated times and samples were taken from the medium to examine IL-8 release. The values in panel A are the means of two experiments. The values for IL-8 mRNA [log2 scale] in panel B are the means ± SEM of the ratios of mRNA at the indicated times to that at the start of the incubation for 5 experiments from as many different individuals. The values in panels B & C are based on the changes in absolute Cp values over time as compared to the Cp value in the unincubated tissue. Statistically significant changes in mRNA are indicated as follows: * P < 0.05 and ** P < 0.025. The IL-8 mRNA values at 1 h and all later times shown in panel B were statistically significant from the value at 0.33 h: P < 0.05. The values in panel C for IL-8, IL-1β, TNFα, NFκB1 and HIF-1α mRNA are from a different series of 4 experiments.

Mentions: The experiments shown in Figure 1 were designed to see how rapid was the upregulation of IL-8 mRNA and protein release by explants of human omental adipose tissue as well as compare IL-8 gene expression at early time points to that of IL-1β, TNFα, NFκB1 [p50 subunit], and HIF-1α mRNA. There was a 8-fold increase in IL-8 gene expression after only 20 minutes incubation of omental adipose tissue explants [Figure 1]. By 2 h, there was a 64-fold increase in IL-8 mRNA. The increase in IL-8 was sustained and reached its highest level by 48 h. The release of IL-8 was also upregulated during incubation, but was not seen until after 40 minutes of incubation and further increases were seen over 48 h. The data in figure 1 also indicate that there were similar increases in IL-1β, TNFα, NFκB1 [p50 subunit], and HIF-1α mRNA within 20 minutes but the increases in the latter two genes were of lesser magnitude.


The inflammatory response seen when human omental adipose tissue explants are incubated in primary culture is not dependent upon albumin and is primarily in the nonfat cells.

Fain JN, Cheema P, Tichansky DS, Madan AK - J Inflamm (Lond) (2010)

Upregulation of the inflammatory response is rapid in onset. Explants of human omental adipose tissue were incubated for the indicated times and samples were taken from the medium to examine IL-8 release. The values in panel A are the means of two experiments. The values for IL-8 mRNA [log2 scale] in panel B are the means ± SEM of the ratios of mRNA at the indicated times to that at the start of the incubation for 5 experiments from as many different individuals. The values in panels B & C are based on the changes in absolute Cp values over time as compared to the Cp value in the unincubated tissue. Statistically significant changes in mRNA are indicated as follows: * P < 0.05 and ** P < 0.025. The IL-8 mRNA values at 1 h and all later times shown in panel B were statistically significant from the value at 0.33 h: P < 0.05. The values in panel C for IL-8, IL-1β, TNFα, NFκB1 and HIF-1α mRNA are from a different series of 4 experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2818611&req=5

Figure 1: Upregulation of the inflammatory response is rapid in onset. Explants of human omental adipose tissue were incubated for the indicated times and samples were taken from the medium to examine IL-8 release. The values in panel A are the means of two experiments. The values for IL-8 mRNA [log2 scale] in panel B are the means ± SEM of the ratios of mRNA at the indicated times to that at the start of the incubation for 5 experiments from as many different individuals. The values in panels B & C are based on the changes in absolute Cp values over time as compared to the Cp value in the unincubated tissue. Statistically significant changes in mRNA are indicated as follows: * P < 0.05 and ** P < 0.025. The IL-8 mRNA values at 1 h and all later times shown in panel B were statistically significant from the value at 0.33 h: P < 0.05. The values in panel C for IL-8, IL-1β, TNFα, NFκB1 and HIF-1α mRNA are from a different series of 4 experiments.
Mentions: The experiments shown in Figure 1 were designed to see how rapid was the upregulation of IL-8 mRNA and protein release by explants of human omental adipose tissue as well as compare IL-8 gene expression at early time points to that of IL-1β, TNFα, NFκB1 [p50 subunit], and HIF-1α mRNA. There was a 8-fold increase in IL-8 gene expression after only 20 minutes incubation of omental adipose tissue explants [Figure 1]. By 2 h, there was a 64-fold increase in IL-8 mRNA. The increase in IL-8 was sustained and reached its highest level by 48 h. The release of IL-8 was also upregulated during incubation, but was not seen until after 40 minutes of incubation and further increases were seen over 48 h. The data in figure 1 also indicate that there were similar increases in IL-1β, TNFα, NFκB1 [p50 subunit], and HIF-1α mRNA within 20 minutes but the increases in the latter two genes were of lesser magnitude.

Bottom Line: In the nonfat cells derived by a 2 h collagenase digestion of omental fat there was an inflammatory response comparable but not greater than that seen in tissue.The inflammatory response was not seen with respect to omentin/intelectin.The marked inflammatory response seen when human omental adipose tissue is incubated in vitro is reduced but not abolished in the presence of albumin with respect to IL-1beta, TNFalpha, IL-8, and is primarily in the nonfat cells of adipose tissue.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Molecular Sciences, College of Medicine, University of Tennessee Health Science Center, Memphis, TN 38163, USA.

ABSTRACT

Background: The present studies were designed to investigate the changes in gene expression during in vitro incubation of human visceral omental adipose tissue explants as well as fat cells and nonfat cells derived from omental fat.

Methods: Adipose tissue was obtained from extremely obese women undergoing bariatric surgery. Explants of the tissue as well as fat cells and the nonfat cells derived by digestion with collagenase were incubated for 20 minutes to 48 h. The expression of interleukin 1beta [IL-1beta], tumor necrosis factor alpha [TNFalpha], interleukin 8 [IL-8], NFkappaB(1)p50 subunit, hypoxia-inducible factor 1alpha [HIF1alpha], omentin/intelectin, and 11beta-hydroxysteroid dehydrogenase 1 [11beta-HSD1] mRNA were measured by qPCR as well as the release of IL-8 and TNFalpha.

Results: There was an inflammatory response at 2 h in explants of omental adipose tissue that was reduced but not abolished in the absence of albumin from the incubation buffer for IL-8, IL-1beta and TNFalpha. There was also an inflammatory response with regard to upregulation of HIF1alpha and NFkappaB1 gene expression that was unaffected whether albumin was present or absent from the medium. In the nonfat cells derived by a 2 h collagenase digestion of omental fat there was an inflammatory response comparable but not greater than that seen in tissue. The exception was HIF1alpha where the marked increase in gene expression was primarily seen in intact tissue. The inflammatory response was not seen with respect to omentin/intelectin. Over a subsequent 48 h incubation there was a marked increase in IL-8 mRNA expression and IL-8 release in adipose tissue explants that was also seen to the same extent in the nonfat cells incubated in the absence of fat cells.

Conclusion: The marked inflammatory response seen when human omental adipose tissue is incubated in vitro is reduced but not abolished in the presence of albumin with respect to IL-1beta, TNFalpha, IL-8, and is primarily in the nonfat cells of adipose tissue.

No MeSH data available.


Related in: MedlinePlus