LACTIC-ACIDOSIS INCREASES TUMOR-NECROSIS-FACTOR SECRETION AND TRANSCRIPTION INVITRO
LACTIC-ACIDOSIS INCREASES TUMOR-NECROSIS-FACTOR SECRETION AND TRANSCRIPTION INVITRO
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DOI：
10.1016/0022-4804(90)90036-2
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发表时间：
1990-10-01
影响因子：
2.2
通讯作者：
NORTON, JA
中科院分区：
文献类型：
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作者：
JENSEN, JC;BURESH, C;NORTON, JA
乳酸已被证明影响许多生物过程。我们研究了乳酸酸中毒作为体外巨噬细胞产生TNF的信号的作用。雄性F344大鼠腹腔内给予巯基乙酸盐培养基。7天后回收巨噬细胞，在完全培养基（CM）或含有L-乳酸（5、10或15 mM）或内毒素（LPS）（10 μ g/ml）的CM中培养24小时。测量上清液中的TNF水平。类似地处理雌性C57 BL/6小鼠，收获巨噬细胞并在CM或含有乳酸（15 mM）或LPS（10 μ g/ml）的CM中培养。24小时后提取RNA，通过电泳分离，并转移至硝酸纤维素。人32 P-cDNA TNF和肌动蛋白探针用于确定相对TNF基因表达。如所述，使用凝胶密度测定法计算乳酸和LPS处理的细胞中的TNF表达指数（EI）。上清液的pH值水平表明，乳酸浓度的增加导致酸中毒增加。台盼蓝拒染法证明乳酸酸中毒不会降低细胞活力。与对照组相比，LPS显著增加TNF的分泌（P < 0.001）。各浓度乳酸均能显著增加TNF的分泌（P < 0.05），但无剂量依赖性。与对照组相比，在用乳酸和LPS培养的巨噬细胞中TNF基因转录升高（EI分别为1.13和1.18）。这表明乳酸浓度可以在转录水平上调节TNF分泌，并且与局部水平的乳酸（乳酸酸中毒）可能是细胞因子分泌的调节剂的假设一致。
Lactic acid has been shown to affect numerous biologic processes. We investigated the role of lactic acidosis as a signal for the production of TNF by macrophages in vitro. Male F344 rats were administered thioglycolate media intraperitoneally. Macrophages were recovered 7 days later, cultured for 24 hr in complete media (CM), or CM with L-lactic acid (5, 10, or 15 mM), or with endotoxin (LPS) (10 .mu.g/ml). TNF levels were measured in the supernatants. Female C57BL/6 mice were similarly treated, and macrophages were harvested and cultured in CM or CM containing lactic acid (15 mM), or LPS (10 .mu.g/ml. RNA was extracted after 24 hr, separated by electrophoresis, and transferred to nitrocellulose. Human 32P-cDNA TNF and actin probes were used to determine relative TNF gene expression. Gel densitometry was used to calculate the TNF expression index (EI) in lactic acid and LPS treated cells as described. pH levels of the supernatant indicated that increasing concentrations of lactic acid caused increasing acidosis. Trypan blue exclusion demonstrated that lactic acidosis did not reduce cell viability. LPS significantly increased secretion of TNF relative to control (P < 0.001). Each concentration of lactic acid significantly increased TNF secretion (P < 0.05), but not in a dose-dependent manner. TNF gene transcription was elevated in macrophages cultured with lactic acid and LPS relative to control (EI = 1.13 and 1.18, respectively). This suggests that lactic acid concentration can regulate TNF secretion at the level of transcription, and is consistent with the hypothesis that local levels of lactic acid (lactic acidosis) may be a regulator of cytokine secretion.