摘要
本试验旨在通过给小鼠腹腔注射脂多糖(LPS),检测随着时间的推移小鼠血液细胞因子水平和肠道组织形态的变化。25只雄性ICR小鼠经腹腔注射LPS(5 mg/kg BW),在不同致炎时间[LPS注射前(0 h)以及注射后3、6、12、24、48和72 h]采血制备血清,通过酶联免疫吸附试验(ELISA)方法检测小鼠血清肠碱性磷酸酶(IAP)活性及炎性细胞因子肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-4、IL-6和IL-8的水平;同时采集小鼠十二指肠、空肠、回肠和盲肠样品,苏木精-伊红(HE)染色制作组织切片,观察各肠段病理学变化,并测量各肠段绒毛高度(VH)和隐窝深度(CD),计算VH/CD(V/C)。结果显示:血清IAP活性,促炎细胞因子TNF-α、IL-6和IL-8以及抗炎细胞因子IL-4的水平在LPS注射后6 h时达到高峰,随后下降;而血清促炎细胞因子IL-1β的水平在LPS注射后6 h时略有升高,在注射后48 h时达高峰。与注射前(0 h)相比,LPS致炎小鼠在LPS注射后6 h肠道病理变化比较明显,小鼠肠黏膜损伤明显,上皮脱落、绒毛破裂、黏膜萎缩、水肿、且绒毛较短;随着时间的推移,肠黏膜在LPS注射后48 h时已开始缓慢恢复。与注射前(0 h)相比,LPS致炎小鼠十二指肠和空肠的V/C均在注射后6 h时出现显著或极显著下降(P<0.05或P<0.01),但在注射后72 h时已无显著差异(P>0.05)。由此得出,LPS(5 mg/kg BW)腹腔注射后6 h时小鼠血清促炎性细胞因子水平达到高峰,肠黏膜损伤明显,之后逐渐恢复正常。
This experiment was conducted to detect the changes in blood cytokine levels and intestinal tissue morphology of mice by intraperitoneal injection of lipopolysaccharide( LPS). Twenty-five male ICR mice were intraperitoneally injected with LPS( 5 mg/kg BW),and the blood were sampled to prepare serum at different time points of inflammation [before LPS injection( 0 h) and 3,6,12,24,48 and 72 h after injection]. The activity of intestinal alkaline phosphatase( IAP) and the levels of inflammatory cytokine,such as tumor necrosis factor-α( TNF-α),interleukin( IL)-1β,IL-4,IL-6 and IL-8 in serum of mice were detected by enzymelinked immunosorbent assay( ELISA) method. The samples of duodenum,jejunum,ileum and cecum were collected from the mice and hematoxylin-eosin( HE) staining was performed to observe the pathological changes of each intestinal segment. The villous height( VH) and crypt depth( CD) in each intestinal segment were measured and the VH/CD( V/C) was calculated. The results showed as follows: as time progressed,the activity of serum IAP and the levels of serum pro-inflammatory cytokines TNF-α,IL-6 and IL-8,and anti-inflammatory cytokines IL-4 peaked at 6 h after LPS injection,and then decreased continuously. The levels of IL-1β,a pro-inflammatory cytokine,increased slightly at 6 h after LPS injection and peaked at 48 h after injection.Compared with 0 h before LPS injection,the intestinal pathological changes of LPS-induced mice at 6 h after LPS injection were obvious. The mucous membranes of intestine were damaged,the epithelium was detached,the villus was broken,the mucous membranes were atrophic,edema,and the villus was short; over time,intestinal mucosa had slowly recovered at 48 h after LPS injection. Compared with 0 h before LPS injection,V/C in the duodenum and jejunum of LPS-inflamed mice were significantly decreased at 6 h after injection( P<0.05 or P<0.01); however,there was no significant difference at 72 h after injection( P>0.05). It is concluded that the levels of serum pro-inflammatory cytokines peaked at 6 h after intraperitoneal injection of LPS( 5 mg/kg BW),intestinal mucosal damage is obvious,and gradually returns to normal afterwards.
引文
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