人参皂甙RB2抑制NF-κB的激活对LPS诱导的新生小鼠急性肺损伤的免疫调节作用

Immunomodulatory effect of ginsenoside RB2 on lipopolysaccharide-induced acute lung injury in neonatal mice by inhibiting NF-κB activating

目的:研究人参皂苷RB2对脂多糖(LPS)诱导的新生小鼠急性肺损伤的免疫功能的影响及相关分子机制。方法:将新生小鼠随机分为四组:健康组(Ctrl);人参皂苷单独处理组(GR2):健康小鼠腹腔注射GR2 50 mg/kg BW; LPS诱导组(LPS):腹腔注射0. 6 mg/kg BW的LPS,连续5 d; LPS+GR2组:模型小鼠腹腔注射人参皂苷50 mg/kg BW。HE染色观察肺组织损伤情况; TUNEL染色观察肺组织细胞凋亡; Western blot检测Caspase-3和Caspase-9的表达; ELISA检测炎症因子IL-6、IL-1β、TNF-α和IL-10的含量; Western blot检测高迁移率族蛋白B1(HMGB1)、巨噬细胞移动抑制因子(MIF)和NF-κBp 65的表达水平。结果:LPS组与对照组相比,肺组织出现明显的损伤,肺泡壁明显增厚,出现大量炎性细胞浸润;染色呈阳性的凋亡细胞比率显著提高; Caspase-3和Caspase-9的表达水平显著上调;炎症因子IL-6、IL-1β和TNF-α的含量显著上升,IL-10的含量没有明显变化; HMGB1和MIF的表达显著上调;磷酸化的NF-κB p65的表达显著上调。LPS+GR2组与LPS组相比,肺组织损伤明显得到缓解,多数肺泡结构完整,仅有少量炎性细胞浸润;凋亡细胞的比率显著下降; Caspase-3和Caspase-9的表达水平显著下调;炎症因子IL-6、IL-1β和TNF-α的含量显著下降,IL-10的含量没有明显的变化; HMGB1和MIF的表达显著下调;磷酸化的NF-κB p65的表达显著下调。结论:人参皂苷RB2抑制NF-κB的激活,进而调节LPS诱导的新生小鼠急性肺损伤的免疫反应。

Objective:To explore the effects of ginsenoside RB2 on immune function of lipopolysaccharide induced acute lung injury in neonatal mice and its molecular mechanism. Methods: Neonatal mice were randomly divided into four groups:the healthy group (Ctrl) and the ginsenoside alone treatment group (GR2):the healthy mice were intraperitoneally injected with GR2 50 mg/kg BW;LPS induced group (LPS):intraperitoneal injection of 0.6 mg/kg BW LPS for 5 days;LPS+GR2 group:the model mice were intraperitoneally injected with ginsenoside 50 mg/kg BW.HE staining was used to observe the damage of lung tissue;apoptosis of lung tissue was observed by TUNEL staining;expression of Caspase-3 and Caspase-9 was detected by Western blot;ELISA was used to detect the levels of inflammation factor IL-6,IL-1β,TNF-α and IL-10. Results: Compared with the control group,the lung tissue was obviously damaged,and a large number of inflammatory cells were infiltrated.The percentage of apoptotic cells increased significantly;the expression level of Caspase-3 and Caspase-9 increased significantly;the content of inflammatory factors IL-6,IL-1β and TNF-α increased significantly.There was no obvious change in the content of IL-10.The expression of HMGB1 and MIF was significantly up-regulated,while the expression of phosphorylated NF-κB p65 was significantly upregulated.LPS+GR2 group compared with the LPS group,the lung tissue damage was significantly relieved.Most of the alveolar structures were intact,only a small amount of inflammatory cells were infiltrated,the percentage of apoptotic cells decreased significantly,the expression level of Caspase-3 and Caspase-9 decreased significantly,and the content of IL-6,IL-1β and TNF-α decreased significantly,and there was no obvious change in the content of IL-10.The expression of HMGB1 and MIF was significantly down regulated;the expression of phosphorylated NF-κB p65 was significantly downregulated. Conclusion: Ginsenoside RB2 inhibits the activation of NF-κB and further regulates the immune response of LPS induced acute lung injury in neonatal mice.