Changes of Proliferation and Apoptosis Processes in Fibroblasts in Abdominal Adhesions Formation
S.V. Skalsky, T.F. Sokolova
DOI: 10.22138/2500-0918-2017-14-4-369-374
Omsk State Medical University, Omsk, Russian Federation
Abstract. Objective of the study is to evaluate the processes of proliferation, collagenosis and apoptosis in fibroblasts activated in vivo while modeling hemoperitoneum in rats. Material and methods. The studies were performed in the primary cell culture on 114 pools of rat peritoneal fibroblasts obtained from 26 animals with hemoperitoneum on the 10th day after simulating the model; it was a main group, and 12 control rats. A suspension of peritoneal fibroblasts (2×106 cells/ml) was cultured for 24 hours (t37°C, 5% CO2). Proliferative activity was assessed by the number of fibroblasts with mitosis morphological features, as well as by the expression of a proliferation marker Ki-67 using the monoclonal antibodies. The degree of collagenosis stimulation by fibroblasts was evaluated by the amount of protein-bound oxyproline in the supernatants of the cell cultures. Amount of active forms of the nuclear transcription factor NF-kB was determined by ELISA. The revision of the abdominal cavity in rats with macro- and microscopic analysis of adhesions was performed. It was found that the proliferative activity of fibroblasts was high in rats with hemoperitoneum: the mitotic index exceeded the control values by more than 5 times, the level of the proliferation marker Ki-67 was increased almost 8 times. It was detected a 4-fold increase in the collagen-synthetic function of activated fibroblasts. A direct correlation between the proliferative activity of fibroblasts, the amount of protein-bound oxyproline and the number of abdominal adhesions in rats was noted (r=0.91, r=0.78, p<0.05). The intensification of fibroblasts proliferation, their collagen-synthetical function in adhesion process induction in the abdominal cavity was accompanied by the activation of the transcription factor NF-κB, leading to the blocking of apoptosis processes. The indices of activation for NF-κB peritoneal fibroblasts in rats with hemoperitoneum were 8 times higher than the control values. Conclusions. Thus, violations of growth autoregulation of the connective tissue in hemoperitoneum with the predominance of fibroblast proliferation induction, accompanied by activation of their collagen-synthesizing function over inhibition, leading to the adhesion progression.
Keywords: adhesion, proliferation, hydroxyproline, apoptosis, proliferation marker Ki-67, nuclear transcription factor NF-κB
Received 08.09.2017
For citation:
Skalsky S.V., Sokolova T.F. Changes of proliferation and apoptosis processes in fibroblasts in abdominal adhesions formation. Vestn. Ural. Med. Akad. Nauki. Journal of Ural Medical Academic Science. 2017, Vol. 14, no. 4, pp. 369–374. DOI: 10.22138/2500-0918-2017-14-4-369-374 (In Russ)
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Authors
Sergey V. Skalsky
Cand. Sci. (Med.), Docent, Head of the Department of Pharmacology
sergscalskiy@mail.ru
Tatyana F. Sokolova
Doc. Sci. (Med.), Accistant of the Department of Pharmacology
tfsokolova@mail.ru
Omsk State Medical University
Russian Federation, 644099 Omsk, ul. Lenin, 12