背景:急性肺损伤(ALI)是一种由创伤引起的急性多因素感染性疾病,肺炎,震惊,还有败血症.本研究旨在探讨伪麻黄碱和大黄素联合治疗对实验性ALI的保护作用。以及通过VIP/cAMP/PKA途径调节炎症和肺水肿的潜在机制。
方法:wistar大鼠随机分为15组(n=5)。各组大鼠在注射LPS前1h灌胃给药。对照组和LPS组给予生理盐水灌胃,其他组的大鼠胃内给予不同剂量的治疗剂。然后将LPS和治疗组的大鼠腹膜内注射LPS(7.5mg/kg)以诱导ALI。用伪麻黄碱和大黄素治疗12小时后,所有的动物都是祭品。在LPS注射后8小时内每小时测量肛门温度。肺标本病理检查采用H&E染色。细胞因子(IL-1β,TNF-α,IL-6,iNOS,通过ELISA和免疫荧光测定肺组织中的IL-10,Arg-1,CD86,CD206,F4/80,VIP)。VIP的表达,CAMP,AQP-1,AQP-5,p-PKA,PKA,p-IκBα,IκBα,通过蛋白质印迹法测定肺中p-p65,p65,p-P38,P38,p-ERK1/2,ERK1/2,p-JNK1/2,JNK1/2蛋白。
结果:大鼠接受伪麻黄碱+大黄素治疗后,减少发烧症状。炎性细胞因子(IL-1β,TNF-α,血清中IL-6,iNOS)降低,抗炎细胞因子(IL-10,Arg-1)明显升高。伪麻黄碱+大黄素治疗可有效促进肺组织VIPcAMP和p-PKA蛋白表达,并显著抑制NF-κB,MAPK磷酸化,伪麻黄碱+大黄素处理可通过VIP/cAMP/PKA信号通路抑制M1极化,促进M2极化。
结论:伪麻黄碱和大黄素联合应用可通过诱导VIP/cAMP/PKA信号通路改善LPS诱导的大鼠ALI。抑制NF-κB,MAPK炎症通路,缓解肺水肿抑制巨噬细胞M1极化,促进巨噬细胞M2极化。
BACKGROUND: Acute lung injury (ALI) is an acute multifactorial infectious disease induced by trauma, pneumonia, shock, and sepsis. This study aimed to investigate the protective effects of pseudoephedrine and emodin combined treatment in experimental ALI, as well as the mechanisms underlying the regulation of inflammation and pulmonary edema via the
VIP/cAMP/PKA pathway.
METHODS: The wistar rats were randomly divided into fifteen groups (n = 5). Rats in each group were given intragastric administration 1 h before LPS injection. Those in the control and LPS groups were given intragastric administrations of physiological saline, rats in other groups were given intragastrically administered of differential dose therapeutic agents. The rats in the LPS and treatment groups were then injected intraperitoneally with LPS (7.5 mg/kg) to induce ALI. After being treated with pseudoephedrine and emodin for 12 h, all animals were sacrifice. Anal temperatures were taken on an hourly basis for 8 h after LPS injection. Pathological examination of lung specimen was performed by H&E staining. Cytokines (IL-1β, TNF-α, IL-6, iNOS, IL-10, Arg-1, CD86, CD206, F4/80,
VIP) in lung tissue were assayed by ELISA and immunofluorescence. The expression of
VIP, CAMP, AQP-1, AQP-5, p-PKA, PKA, p-IκBα, IκBα, p-p65, p65, p-P38, P38, p-ERK1/2, ERK1/2, p-JNK1/2, JNK1/2 protein in lung was determined by western blotting.
RESULTS: After rats being treated with pseudoephedrine + emodin, reduced of fever symptoms. The contents of inflammatory cytokines (IL-1β, TNF-α, IL-6, iNOS) were decreased and anti-inflammatory cytokines (IL-10, Arg-1) were significantly increased in serum. Pseudoephedrine + emodin treatment effectively promoted
VIP cAMP and p-PKA protein expression in lung tissues, and significantly inhibited NF-κB, MAPK phosphorylation, Pseudoephedrine + emodin treatment can inhibit M1 polarization and promoted M2 polarization via the
VIP/cAMP/PKA signaling pathway.
CONCLUSIONS: The combination of Pseudoephedrine and emodin was effective in ameliorating LPS-induced ALI in rats by inducing VIP/cAMP/PKA signaling. Inhibiting the NF-κB, MAPK inflammatory pathway, relief of pulmonary edema suppressing macrophage M1 polarization, and promoting macrophage M2 polarization.