Effects of Pancragen on The Differentiation of Pancreatic Cells During Their Ageing

Abstract

Expression of differentiation markers was found to be reduced during ageing of pancreatic cells. Tetrapeptide pancragen stimulates the expression of differentiation factors of acinar (Pdx1, Ptfla) and islet of Langerhans (Pdx1, Pax6, Pax4, Foxa2, NKx2.2) cells in "young" and " aged" cultures. Differentiation of acinar and islet pancreatic cells induced by pancragen can be a mechanism underlying its anti-diabetic and anti-inflammatory effects. Thus, transcription factors that regulate differentiation of pancreatic cells are a pharmacological target for pancragen, which allows considering it as an effective tool in the treatment of diabetes mellitus and pancreatitis.

Tetrapeptide KEDW Interacts with DNA and Regulates Gene Expression

“…An increase in cell culture passages number results in decreased gene expression levels of differentiation factors; this is necessary in order to induce maturation of islet pancreatic cells [15]. It is understood that absence/depression of synthesis of these proteins results in a disturbed development of pancreatic cells from pluripotent cells [6].…”

“…However, we assume that peptide regulation of gene expression existing under physiological conditions should not lead to overexpression of genes. Even an increase of genes expression 2-3 times by peptide KEDW promotes the synthesis of pancreatic differentiation factor regulators by 2-3 times, which leads to the restoration of pancreatic function, as shown in animal studies [13][14] [15]. We could assume that if the expression of genes and the synthesis of the corresponding proteins increased tenfold it would lead to short-term improvement of pancreatic function and rapid resource depletion with long-term negative consequences.…”

“…Cell cultures were divided into 2 equal groups: to the first group, we added 165 mL of normal saline solution and to the second group, the same 165 mL solution and additional KEDW peptide (20 ng/mL). This concentration was used as it seemed the most effective in previous studies [15] [14]. Cells were grown in vials with 25 cm surface (JetBiofil, Japan) in 5 mL of growth DMEM medium at 37ºC with addition of L-glutamine (Biolot), 15% of fetal bovine serum SC-BIOL (Biolot), and 1% PenicillinStreptomycin solution.…”

“…Mutations in these differentiation and expression genes can disturb insulin synthesis in β-cells and lead to DM [7][8] [9][10] [11] [12]. Expression of differentiation and proliferation factors, which regulate the maturation of endocrine insular cells and exocrine acinar, gradually decreases with continuous propagation of pancreatic cell cultures [13][14] [15].…”

“…KEDW peptide is also shown to lower blood glucose after fasting and during standard glucose tolerance tests; additionally, KEDW peptide can lower the concentration of plasma insulin and the insulin sensitivity index in patients of advanced age with DM types 1 and 2 [17]. Furthermore, KEDW peptide increases synthesis of Pax6, Foxa2, Nkx2.2, Nkx6.1, Hoxa3, and Pax4 factors in cell cultures at the 14 th passage [13][14] [15]. Herein, the project aim is to study the mechanism of KEDW peptide influence on gene expression of aforementioned transcription factors in human pancreatic cell cultures.…”