目的 探讨多黏菌素B(PMB)干预颅脑创伤后大鼠外周血肠道细菌崩解产物脂多糖(LPS)表达的变化。方法 选取健康7周龄成年雄性SD大鼠50只,随机分为5组,正常组(Normal)、假手术组(Sham)、创伤性脑损伤组(TBI)、创伤性脑损+生理盐水组(TBI+ NS组)及创伤性脑损+多黏菌素组(TBI+PMB组),每组10只。通过电子皮质损伤撞击仪(eCCI)造模, 分别计算Normal、Sham、TBI组大鼠造模前后24、48 h神经功能缺损评分(NSS)。造模后48 h,这三组大鼠给予FD4灌胃,检测循环中FD4的浓度间接反映肠道通透性改变;TBI+PMB组使用1 ml注射器经腹腔注射PMB溶液(2.5 μg/ml),TBI+ NS组给予等体积生理盐水。记录实验前,试验第1、3、5、7 天 共5个时间点的(试剂终点显色法检测)外周血中LPS的浓度;并结合7 d脑组织(石蜡包埋HE染色)炎症评分评估脑组织炎症反应。结果 与Sham组比较, Normal组外周血中FD4的浓度升高[(2743±279)ng/ml vs. (1850±80) ng/ml)],TBI组外周血中FD4的浓度明显增高[(4228±203)ng/ml vs. (2743±279)ng/ml],LPS浓度在第3天时显著升高,差异均有统计学意义(P<0.05)。其余时间点差异无统计学意义。与TBI+NS组比较,TBI+PMB组LPS浓度在术后第3天明显降低[(0.56±0.04)EU/ml vs.(0.94±0.03)EU/ml];炎症评分TBI组和TBI+NS组明显下降,差异均有统计学意义(P<0.05)。结论 大鼠急性TBI后可出现肠道菌群移位及外周血LPS水平一过性增加,PMB能降低TBI后早期LPS浓度,减少急性TBI后脑组织炎症反应。
Abstract
Objective To explore the effect of polymycin B (PMB) on the expression of lipopolyssacharide (LPS)in peripheral blood of rats after craniocerebral trauma. Methods Fifty healthy 7-week-old adult male SD rats were randomly divided into 5 groups: normal group, sham group, traumatic brain injury group (TBI), traumatic brain injury+Normal saline group (TBI+NS group), traumatic brain injury + polymycin group (TBI+PMB group), with 10 rats in each group. Electronic cortical injury impingement instrument (eCCI) was used to construct models. The neurological deficit scores (NSS) of the rats in the first 3 groups were calculated at 24 h and 48 h before and after modeling. At 48h after modeling, the rats in the first three groups were given FD4 gavage, and the concentration of FD4 in the cycle was detected to indirectly reflect changes in intestinal permeability. The rats in TBI+PMB group were intraperitoneally injected with PMB solution (2.5 μg/g) through 1 ml syringe, and those in TBI+ NS group were given equal volume of normal saline. At the time points of 0 d, 1 d, 3 d, 5 d and 7 d, the concentration of LPS in peripheral blood was detected by limulus lysate end-point colorimetric method. Paraffin-embedded HE staining and inflammation scores were used to evaluate the inflammatory response of brain tissue at 7 d. Results Compared with the sham group[(1850±80) ng/ml], the concentration of FD4 in the normal group and the TBI group increased [(2743±279) ng/ml vs. (4228±203) ng/ml](P<0.05); compared with the sham group, LPS concentration in TBI group significantly increased at day 3 [(0.94±0.02) EU/ml vs. (0.45±0.06) EU/ml], and there was no significant difference at other time points. Compared with TBI+NS group, LPS concentration in TBI+PMB group significantly decreased at day 3 after operation [(0.56±0.04) EU/ml vs. (0.94±0.03) EU/ml, P<0.001];inflammation scores in TBI+PMB group were significantly lower than those in TBI group and TBI+NS group. Conclusions The acute TBI can cause intestinal flora shifting and transient increase of LPS level in peripheral blood of rats. PMB can reduce the early LPS concentration after TBI and reduce the inflammatory response in brain tissues after acute TBI.
关键词
多黏菌素B /
创伤性脑损伤 /
肠道细菌崩解产物脂多糖
Key words
polymycin B /
traumatic brain injury /
lipopolyssacharide
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