Korean J Gastroenterol.  2019 Oct;74(4):219-226. 10.4166/kjg.2019.74.4.219.

Screening of Sera from Patients with Pancreatitis by an Apoptosis Assay of Skin-derived Cells

• Seok AE
• Son BK
• Lee J
• Chung KH
• Lee YR
• Kim D
• Cha BH
• Kang HG

• KMID: 2460987
• DOI: http://doi.org/10.4166/kjg.2019.74.4.219

Abstract

BACKGROUND/AIMS
An excessive inflammatory response is typical in acute pancreatitis and a significant cause of early mortality in severe acute pancreatitis. This is believed to be caused by inflammatory molecules or upregulated cytokine levels in the serum of patients. The aim of this study was to identify the serum-mediated apoptosis-inducing effects in acute pancreatitis patients.
METHODS
A skin tissue-derived cell line, BJ, was treated for 24 hours with the sera of 22 healthy volunteers (control) and 71 acute pancreatitis patients (22 with gallstone pancreatitis, 16 with alcoholic pancreatitis, and 11 with pancreatitis with other causes) collected at the time of hospital admission (active) and discharge (resolved). Apoptosis was analyzed by flow cytometry.
RESULTS
The average percentage of living cells, early apoptotic cells, and late apoptotic cells ranged from 78.8% to 85.0%, 5.5% to 7.3%, and 7.7% to 13.1%, respectively. The number of live cells increased significantly using the serum from the resolved state of gallstone-induced pancreatitis. In addition, the number of early apoptotic cells increased significantly using the serum from the resolved state of pancreatitis with other causes. The number of late apoptotic cells decreased significantly with the serum from the resolved state compared to the active state of gallstone- and alcohol-induced pancreatitis.
CONCLUSIONS
Serum samples from patients with pancreatitis induced a change in the apoptosis profiles of skin-derived cells. These results indicate changes in the serum components in patients with acute pancreatitis.

Keyword

Acute pancreatitis; Skin tissue-derived cell line; Apoptosis; Alcohol-induced pancreatitis

MeSH Terms

Apoptosis*
Cell Line
Flow Cytometry
Gallstones
Healthy Volunteers
Humans
Mass Screening*
Mortality
Pancreatitis*
Pancreatitis, Alcoholic
Skin

Figure

• 

  Fig. 1 Apoptosis analysis of sera from healthy control and acute pancreatitis patients of BJ cell lines. Scatter plot of Annexin V and propidium iodide staining for healthy control serum- and patient serum-treated cells. Quantitative analysis of the percentage of live cells, and the early and late apoptotic cells by flow cytometry are shown. Q, quadrant; UL, upper left; UR, upper right; LL, lower left; LR, lower right.

• 

  Fig. 2 Flow cytometry analysis of sera from patients with acute pancreatitis includes apoptosis of BJ cell lines. Scatter plot of Annexin V and propidium iodide staining acute pancreatitis patient serum-treated cells. Quantitative analysis of the percentage of live cells, and early and late apoptotic cells by flow cytometry analysis are shown. Error bars, mean±standard error. AS, active state; RS, resolved state; Q, quadrant; UL, upper left; UR, upper right; LL, lower left; LR, lower right.

• 

  Fig. 3 Comparative analysis of living cells in an apoptosis assay. Error bars, mean±standard error. *p-value <0.05.

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  Fig. 4 Comparative analysis of early apoptotic cells in an apoptosis assay. Error bars, mean±standard error. *p-value <0.05.

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  Fig. 5 Comparative analysis of late apoptotic cells in an apoptosis assay. Error bars, mean±standard error. *p-value <0.05.

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