鼠源重组UBC13蛋白对脂多糖诱导的小鼠急性炎症的影响

试验旨在探讨鼠源重组UBC13蛋白对脂多糖(lipopolysaccharide,LPS)诱导的小鼠急性炎症的影响。将24只SPF雌性小鼠随机分成4组:PBS组,LPS模型组,重组UBC13蛋白高、低剂量组(分别为100和25μg/只),每组6只。LPS模型组与各蛋白剂量组腹腔注射20 mg/kg LPS,PBS组腹腔注射等体积PBS;注射结束1 h后,各蛋白组按相应剂量背部皮下多点注射重组UBC13蛋白,PBS组与LPS模型组注射等体积PBS。给予蛋白24 h后处死小鼠。收集小鼠肺脏、脾脏、胸腺及肝脏组织,计算脏器指数,HE染色观察组织病理学变化,实时荧光定量PCR检测肺脏、脾脏、胸腺和肝脏中IL-1β、TNF-α、IL-6 mRNA的相对表达量,以及肺脏中iNOS mRNA的相对表达量,综合评价鼠源重组UBC13蛋白对LPS诱导小鼠急性炎症的影响。结果显示,与PBS组相比,LPS模型组小鼠肺脏、脾脏及肝脏指数均显著或极显著升高(P<0.05;P<0.01),且肺脏、脾脏和肝脏组织均出现病理变化。实时荧光定量PCR结果显示,与PBS组相比,LPS模型组肺脏、脾脏、胸腺和肝脏中IL-1β、TNF-α、IL-6 mRNA相对表达量均极显著升高(P<0.01),肺脏中iNOS mRNA相对表达量也极显著升高(P<0.01);与LPS模型组相比,UBC13蛋白高剂量组肺脏、脾脏和肝脏中病理变化明显改善,肺脏、肝脏、脾脏中IL-1β、TNF-α、IL-6及肺脏中iNOS mRNA表达量均极显著降低(P<0.01);胸腺中TNF-αmRNA表达量显著降低(P<0.05),IL-6 mRNA和IL-1β表达量极显著降低(P<0.01)。表明鼠源重组UBC13蛋白可下调炎性因子的表达,从而改善LPS诱导的小鼠急性炎症反应。

Effect of Murine Recombinant UBC13 Protein on Acute Inflammation Induced by LPS in Mice

To investigate the effect of murine recombinant UBC13 protein on lipopolysaccharide (LPS)-induced acute inflammation in mice,twenty-four SPF female mice were randomly divided into 4 groups:PBS group,LPS model group,UBC13 high and low dose groups (100 and 25 μg per mouse,respectively),with 6 mice in each group.LPS model group and each protein dose group were intraperitoneally injected with 20 mg/kg LPS,and the PBS group was intraperitoneally injected with equal volume of PBS.After 1 h of injection,each protein group was injected subcutaneously with recombinant UBC13 protein at the corresponding dose,PBS group and LPS model group were injected equal volume PBS.The mice were sacrificed 24 h after the administration of the protein.The lung,spleen,thymus and liver tissues of the mice were collected,the organ indexes were calculated.The histopathological changes were observed by HE staining and the relative expression of IL-1β, TNF-α and IL-6 mRNA in lung,spleen,thymus and liver and iNOS mRNA in lung were detected by Real-time PCR.The results showed that compared with PBS control group,the indexes of lung,spleen and liver in LPS model group were significantly or extremely increased (P<0.05; P<0.01),and the lung,spleen,thymus and liver tissues showed pathological changes.The results of Real-time PCR showed that the relative expressions of IL-1β,TNF-α and IL-6 mRNA in lung,spleen,thymus and liver of LPS model group were extremely significantly increased compared with PBS control group (P<0.01).The relative expression of iNOS mRNA in lung was also extremely significantly increased (P<0.01).Compared with LPS model group,the pathological changes in lung,spleen and liver were significantly improved in high dose group of UBC13 protein;The expressions of IL-1β,TNF-α and IL-6 mRNA in lung,liver and spleen were extremely significantly decreased (P<0.01),and the expression of iNOS mRNA in lungs was also extremely significantly decreased (P<0.01);The expression of TNF-α mRNA in thymus was significantly decreased (P<0.05),the expression of IL-6 mRNA and IL-1β were extremely significantly decreased (P<0.01).It indicated that murine recombinant UBC13 protein could down-regulate the expression of inflammatory factors and alleviate LPS-induced acute inflammatory response in mice.