The Eflect Of Trypsin On The Production Of Cytokines (TNF-α,IL-1β And IL-6) From Monocytes In Vitro

Objective: To study the effect of the trypsin on the production of cytokines (TNF- a JL-1 3 and IL-6) by monocytes in vitro. Based on the relationship between trypsin and the discharge of cytokines, discussing about the mechanism of cytokine discharge in AP.Method: The monocytes were gained from peripheral blood by centrifuge, and the concentration adjusted to 2.5X 105/ml. 1.Everybody's monocytes was divided into nine groups based on the varities of trypsin with each piece of monocytes 250ul/ml.The monocytes were incubated in RPMI medium +10% fetal bovine serum (FBS) at 37'C for 4.5-5.0 hours. The PH of medium was adjust to 7.40-7.45.Supernatants were collected after defined incubation periods and assayed for cytokines (TNF-a JL-1 3 and IL-6). 2. Everybody's monocytes was divided into nine groups based on the varities of trypsin-challenging periods with each piece of monocytes 250ul/ml.The monocytes were incubated with defined concentration of trypsin in RPMI medium +10% fetal bovine serum (FBS) at 37 .The PH of medium was adjust to 7.40-7.45.Supernatants were collected at defined incubation periods and assayed for cytokines(TNF- a or IL-6). 3.Control groups: The monocytes were incubated with trypsin, heat-treated trypsin, or natural saline (NS). Supernatants were collected after defined incubation periods and assayed for cytokines, while the amount of monocytes was counted.Results: l.The monocytes incubated with trypsin displayed an elevation of TNF- a . 1.1 The monocytes incubated with various concentration of trypsin displayed a stepwise dose-dependent increase in TNF- a elevation, which peaked in response to 15ug/ml trypsin (mean 234.15 ?45.42pg/ml)(p<0.01) and then declined. 1.2.The monocytes incubated with the concentration of 15ug/ml trypsin displayed a time-dependent increase in TNF- a elevation that peaked at 20 hours (mean 421.25 ?8.35 )(p<0.05) and then declined. 2.The monocytes incubated with various concentration of trypsin displayed a stepwise dose-dependent increase in IL-1 3 elevation, which peaked in response to 15ug/ml trypsin (mean 202.7 + 25.97pg/ml)(p<0.05) and then declined. 3. The monocytes incubated with trypsin displayed an elevation of IL-6. 3.1 The monocytes incubated with various concentration of trypsin displayed a stepwise dose-dependent increase in IL-6 elevation, which peaked in response to 15ug/ml trypsin (mean 751 ?0.2pg/ml)(p<0.01) and then declined. 3.2 The monocytes incubated with the concentration of 20ug/ml trypsin displayed a time-dependentincrease in IL-6 elevation that peaked at 24 hours (mean 2026.8+119.8)(p<0.01) and then declined. 4 The group of monocytes incubated with trypsin displayed a significant difference in TNF- a elevation compared to the control groups (p<0.05),while there was no significant difference in the amount of monocytes survivities between them (p>0.05)..Conclude: l.In vitro trypsin induced production of TNF-a 5IL-1 3 and IL-6 by monocytes,which displayed a stepwise dose-dependent increase and peaked in response to 15ug/ml or 20ug/ml trypsin then declined. 2. hi vitro trypsin induced production of TNF- a and IL-6 by monocytes, which displayed a time-dependent elevation and peaked at 20 or 24 hours then declined.