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目的 观察内毒素血症时肝组织和游离肝细胞中ＣＤ１４基因和ＣＤ１４蛋白的表达。方法 经尾静脉注入脂多糖（ＬＰＳ， Ｅ ｃｏｌｉ Ｏ１１１．Ｂ４）建立大鼠急性内毒素血症动物模型，并用原位胶原酶灌注法分离肝细胞。用流式细胞仪（ＦＣＭ）测定异硫氢酸荧光素（ＦＩＴＣ）－ＣＤ１４阳性肝细胞数及其荧光强度。同时用逆转录－ＰＣＲ和Ｗｅｓｔｅｒｎｂｌｏｔ法测定肝组织和肝细胞中ＣＤ１４ｍＲＮＡ和ＣＤ１４蛋白的表达。结果 ＦＣＭ显示：内毒素血症大鼠６ｈ和１２ｈＦＩＴＣ－ＣＤ１４阳性细胞数明显增多，荧光强度也明显增加。ＲＴ－ＰＣＲ显示，肝组织和肝细胞中ＣＤ１４ｍＲＮＡ的表达在３ｈ时明显增强，６ｈ达高峰，２４ｈ恢复正常水平；Ｗｅｓｔｅｒｎ ｂｌｏｔ分析示肝组织和肝细胞中ＣＤ１４蛋白的表达在６ｈ明显增高，１２ｈ达高峰，２４ｈ仍有一定表达，各时相点间比较有显著差别（Ｐ＜０．０１）。结论 内毒素血症时能明显上调肝细胞ＣＤ１４基因和ＣＤ１４蛋白的表达。
Objective To observe the expression of CD14 and CD14 in liver and free hepatocytes during endotoxemia. Methods Animal models of acute endotoxemia in rats were established by injecting lipopolysaccharide (LPS, E. coli O111.B4) via tail vein, and hepatocytes were isolated by in situ collagenase perfusion. Flow cytometry (FCM) was used to determine FITC-CD4 positive hepatocytes and their fluorescence intensity. At the same time, the expression of CD14mRNA and CD14 protein in liver and liver cells were determined by RT-PCR and Western blot. Results FCM showed that the number of FITC-CD14 positive cells in endotoxemia rats increased significantly at 6h and 12h, and the fluorescence intensity also increased significantly. The expression of CD14 mRNA in liver tissue and hepatocytes increased significantly at 3h and peaked at 6h, and returned to normal level at 24h. Western blot analysis showed that the expression of CD14 protein in liver and hepatocytes increased significantly at 6h and reached the peak at 12h Peak, 24h still have some expression, the time points were significantly different (P <0.01). Conclusions Endotoxemia can significantly upregulate the expression of CD14 and CD14 in hepatocytes.