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【目的】探讨上调pellino-1在Kupffer细胞（KC）内毒素耐受时对肿瘤坏死因子受体相关因子3（TRAF3）泛素化、 促分裂原活化蛋白激酶（MAPK）信号通路及下游细胞因子分泌情况的影响。【方法】分离、培养 C57BL/6 小鼠 KC，随机分 为2组：①空载对照组：转染空载质粒48 h后，先给予小剂量LPS（10 ng/mL）刺激24 h，再给予大剂量LPS（300 ng/mL）刺激。 ②过表达组：过表达pellino-1慢病毒转染48 h后，先给予小剂量LPS（10 ng/mL）刺激24 h，再给予大剂量LPS（300 ng/mL）刺 激。两组分别于处理后0、5、10、30、60 min收获细胞，蛋白质免疫印迹试验（Western blot）检测pellino-1、K48泛素化（K48 ubiquitin，K48-Ub）、TRAF3、JNK 、p-JNK、p38、p-p38蛋白水平表达变化；酶联免疫吸附法（ELISA）检测细胞培养上清液中 IL-1β、TNF-α及IL-10的分泌情况。【结果】与空载对照组相比，过表达组pellino-1蛋白表达量在各个时间点均明显升高； K48-Ub水平明显升高；TRAF3蛋白表达量明显降低；JNK、p38蛋白表达量没有明显变化，但p-JNK及p-p38蛋白表达量显 著升高；过表达组细胞上清中IL-1β及TNF-α的量明显升高（P < 0.05），而IL-10的量则明显降低（P < 0.05）。【结论】过表达 pellino-1可促进TRAF3蛋白K-48泛素化降解，导致TRAF3蛋白表达量降低，激活下游的MAPK信号，从而抑制内毒素耐受 的形成。

To address the inaccuracy of the Thornhill-Craver method in predicting gas-lift valve flow rates,research on gas-lift valve flow was conducted using air as the medium on a gas-lift valve test platform. Experiments were carried out on gas-lift valves with orifice sizes of 2. 8, 3. 2, 4. 8 and 6. 4 mm,resulting in a functional relationship between valve stem travel and flow coefficient,as well as critical pressure ratio for different orifice sizes. By considering the dynamic changes in surface pressure of the valve ball under operating conditions and introducing the degree of valve opening, the method for solving the valve stem stroke was optimized. A model for the flow area of the gas-lift valve was developed,and a new method for calculating the flow area was derived using known parameters. Based on experimental and theoretical analysis,a new model for predicting the flow rate of gas-lift valves was established. The research results indicate that the new model accurately predicts the flow rates of gas-lift valves for different orifice sizes and under various working conditions,with an average relative error of 3. 34%,which is a significant improvement over the Thornhill-Craver method. This is of great importance for determining the working state of gas-lift valves downhole and for improving the precision of gas-lift design. [ABSTRACT FROM AUTHOR]

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