Studies on the Production of Nitric Oxide, Interleukin-1 and Interleukin-6 in Mice Exposed with Sarcoma 180 Cells

Kim, Kang Ryune; Kim, Kwang Hyuk; Chun, Yong Huck; Sunwoo, Yangil

Abstract

This study was conducted to elucidate the biological role of the overproduced nitric oxide (NO), interleukin-l (IL-l), and interleukin-6 (IL-6) which are known to elicit inflammation, rheumatic arthritis, fever, septic shock or other fatal reactions. I investigated whether the Scarcoma 180 cells elicit NO, IL-l and IL-6 production in vivo and in vitro by measuring the NO, IL-1 and IL-6 level in the splenocyte adherent cell (AD), non-adherent cell (NAD) and whole cell (W) exposed to sarcoma 180 cells. I also measured the NO, IL-1 and IL-6 level in the plasma and peritoneal fluid of the sarcoma 180 cell-transplanted mice after 2 hours, 4 hours, 6 hours and 24 hours of incubation. 1. In the splenocyte exposed to sarcoma 180 cells, the NO production of AD and NAD increased after 2, 4, and 6 hours but decreased after 24 hours of incubation. In the whole cell, the NO production was variable; it showed increased level of synthesis at 2 hours, decreased level at 4 hours, increased level again at 6 hours, and decreased level at 24 hours of incubation. In the plasma of the sarcoma 180 cell-transplanted mice, the NO synthesis significantly increased from the 6 hours of incubation. In the peritoneal fluid, the NO production significantly increased until the 4 hours of incubation then decreased gradually. However, it showed higher level of NO production compared to the control group. 2. For the splenocyte AD cell exposed to saracoma 180 cells, the IL-1 level decreased after 6 hours of incubation. For the W cells, the IL-1 level decreased at 4 hours then increased until 24 hours of incubation. The NAD cell showed increased level of IL-I production from 2 to 24 hours. All these cells showed significantly increased level of IL-1 production compared to the control. In the plasma of the sarcoma 180 cell-transplanted mice, the IL-1 production increased more than twice the level of control from the beginning. In the peritoneal fluid, no IL-1 production was detected as in the control. 3. The IL-6 synthesis of the sarcoma 180 cell-exposed splenocyte singnificantly increased compared to the control: the AD cell showed increased level of IL-6 production after 4 hours of incubation. For the NAD cell, increased level of IL-6 was detected at 2 hours after the incubation. In the plasma of the sarcoma 180 cells transplanted mice, the IL-6 level at 2 hours after incubation was 64.22+/- 5.85 pg/ml. After 4 hours of incubation, the level decreased to 43.55+/-1.56 pg/ml. At six and 24 hours after the incubation, no IL-6 was detected. In the control, IL-6 production was not detected. In the peritoneal fluid, the IL-6 production level was 712.41+/-4.27 pg/ml after 2 hours and 225.71+/-9.74 pg/ml after 4 hours of incubation, producing singnificantly higher level of IL-6 compared to the control. After 6 hours of incubation, IL-6 level was 8.27+/-0.78 pg/ml. After 24 hours of incubation, it decreased to 1.38+/-0.39 pg/ml as time proceeds. After 24 hours of incubation, the IL-6 level was the same as the control. These results suggest that the mice which were exposed to sarcoma 180 cells, nitric oxide, interleukin-1 and interleukin-6 which may lead to inflammation, fever, sepsis and septic shock. This study also helps us better understanding of the role of cytokines in inflammatory reaction.