摘要
目的建立脂多糖（LPS）诱导潘氏细胞脱颗粒的动物模型，研究NF-κB信号通路是否参与脱颗粒过程及其可能作用机制。方法雄性KM小鼠18只，均分为对照组、脂多糖（LPS）组和吡咯烷二硫基甲酸盐（PDTC）组。各组小肠插管，对照组灌注生理盐水，LPS组和PDTC组灌注LPS（100gg／ml，1．5mt／h），PDTC组术前1h腹腔注射NF-κB抑制剂PDTC。收集0—30min、30～60min、60～120min三个时间段灌注液，ELISA检测灌注液溶菌酶浓度并计算相对浓度。免疫组化观察肠黏膜NF-κB活性。结果①各时间段LPS组溶菌酶相对浓度较对照组显著升高伊〈0．05）。②PDTC组溶菌酶相对浓度低于LPS组，至60～120min差异有统计学意义（P〈0．05）。③免疫组化结果显示：NF-κB在肠腺中上部上皮细胞、绒毛上皮细胞、黏膜固有层细胞的胞浆和（或）胞核呈强阳性染色，潘氏细胞胞浆、细胞核呈弱阳性染色。PDTC组染色明显减弱。结论LPS可诱导潘氏细胞脱颗粒，NF-κB可能参与这一过程，其机制可能涉及到间接途径，即LPS首先活化邻近细胞的NF-κB通路再间接引起潘氏细胞脱颗粒。
Objective To establish an experimental model of lipopolysaccharide （LPS）-induced degranulation of Paneth cells, and to investigate the role of NF-κB pathway and its possible mechanism. Methods Eighteen male KM mice were randomly divided into control, LPS and PDTC groups. In each group, the small intestines were intubated. The control group were perfused with physiological saline. The experimental groups were perfused with LPS （100 μg/ml, 1.5 ml/h）and were injected intraperitoneally with saline or NF-κB inhibitor PDTC one hour before surgery. Then the perfusates were collected in three periods （0-30 min, 30-60 min, 60-120 min）. Lysozyme concentration in perfusates were tested by ELISA. The relative concentrations were expressed as the ratio of lysozyme concentration to basal levels. NF-κB activity in intestinal mucosa was observed by immunohistochemistry. Results （1） Relative concentration of lysozyme in LPS group was significantly higher than that in control group at all time duration （P〈0.05）. （2） Relative concentration of lysozyme in PDTC group was lower than that in LPS group. The difference in 60-120 min was statistically significant （P〈0.05）. （3） Immunohistochemistry showed that, in LPS group, strong positive staining of NF-κB p65 was observed in the cytoplasm and （or） nuclei, including epithelial cells in the middle and upper crypts, villous epithelial cells and lamina propria cells. Weak positive staining was observed only in Paneth cells. The staining intensity of the PDTC group were significantly weaker than that in other groups. Conclusion LPS can induce Paneth cell degranulation. NF-κB signaling pathway may be involved in this process. NF-κB signaling pathway may act as an indirect way, i.e. LPS first activates the cells beyond the crypt bottom through the NF-κB pathway, then leads to Paneth cell degranulation by yet unknown mechanisms.
出处
《实验动物与比较医学》
CAS
2010年第2期109-112,共4页
Laboratory Animal and Comparative Medicine
关键词
潘氏细胞
NF-ΚB
脂多糖
脱颗粒
模式识别受体
Paneth cell
NF-κB
Lipopolysaccharide
Degranulation
Pattern recognition receptor
