摘要
目的 探讨烫伤大鼠脾脏和肠系膜淋巴结（ｍｅｓｅｎｔｅｒｉｃ ｌｙｍｐｈ ｎｏｄｅｓ，ＭＬＮｓ）淋巴细胞凋亡与肠道 ＬＰＳ移位的关系。方法 ３０％ＴＢＳＡ Ⅲ度烫伤 Ｗｉｓｔａｒ大鼠及对照组，取门、腔静脉血用鲎基质显色法定量 ＬＰＳ；脾脏和 ＭＬＮｓ切片以 ＴＵＮＥＬ－ＰＯＤ法染色，应用 Ｃｏｏｌｄｃｃｄ成象系统、Ｓｐｏｔ和 ＩＰＰ（Ｉｍａｇｉｎ ｐｒｏｐｌｕｓ）４．１０ａ软件进行凋亡细胞计数和透射光光密度分析。结果 烫伤后大鼠门、腔静脉血ＬＰＳ水平显著增加（Ｐ＜０．０１），伤后６ｈ达峰值；门静脉ＬＰＳ３－４８ｈ显著高于腔静脉（Ｐ＜０ ．０１）。ＴＵＮＥＬ－ＰＯＤ染色结果，ＭＬＮｓ伤前几乎未见凋亡淋巴细胞，但伤后显著增多（Ｐ＜０．０１），以３ｈ为明显；脾脏伤前凋亡淋巴细胞多集中在皮质。伤后显著减少（Ｐ＜０．０１），２４ｈ后略有恢复。透射光光密度值分析表明，伤后２４ ｈ内脾脏凋亡淋巴细胞的光密度值较伤前明显降低（Ｐ＜０．０１）；伤后 ４８ ｈ内 ＭＬＮｓ凋亡淋巴细胞光密度值较伤前显著增高（Ｐ＜０．０１）。结论 大鼠严重烫伤早期即发生肠源性ＬＰＳ血症；肠道移位ＬＰＳ可诱导ＭＬＮｓ淋巴细胞大量凋亡，脾脏中凋亡淋巴细胞数量显著减少，提示?
Objective To investigate the relationship between lymphocyte apoptosis in spleen and mesenteric lymph nodes (MLNs) and gut -origin lipopolysaccharide ( LPS) translocation after severe scal- ding in rats. Methods Wistar rats inflicted 30% TBSA H degree scalding were employed as the model. Serum LPS levels in portal vein and vena cava were determined by limulus amebocyte lysate(LAL) test. Ap- optotic lymphocytes in paraffin - embedded spleen and MLNs sections were detected with TdT - mediated and dUTP nick end labeling (TUNEL) and peroxidase (POD) staining. The images were taken by cooldccd cam- era system,and the count and optical density (OD) value of apoptotic lymphocytes were analyzed with soft- ware Spot and Imagine proplus 4. 10a( IPP4. 10a). Results Serum LPS levels in portal vein and vena ca- va increased sharply in rats postburn ( P < 0. 01) and reached a peak value at 6 postburn hour( PBH). LPS level in portal vein was higher than that in vena cava ( P <0. 01) in the period of 3 to 48 PBHs. The results of TUNEL - POD staining and the count of apoptotic lymphocytes showed that the apoptotic cells were few in normal MLNs, but increased obviously postburn ( P <0.01) , especially at 3 PBH. On the contrary, apop- totic lymphocytes were concentrated in spleen cortex before injury and decreased obviously postburn ( P <0. 01) , though a slight recovery was found after 24 PBH. Optical density of apoptotic lymphocytes was signifi- cantly reduced in spleen at 24 PBH, But the OD value of apoptotie lymphocytes in MLNs increased signifi- cantly within 48 PBHs when compared with that before injury ( P < 0. 01). Conclusion Gut - origin en- dotoxemia in rat could occur in early postburn period. The LPS translocation might induce lymphocyte apop- tosis in MLNs. The decreased apoptotic lymphocytes in spleen implied abnormal immune function in spleen.
出处
《中华烧伤杂志》
CAS
CSCD
2001年第4期228-230,共3页
Chinese Journal of Burns
基金
国家重点基础研究发展规划资助项目(G199905403)
