甘氨酸偶联皮质醇抗Compound 48/80刺激大鼠急性过敏反应的作用研究

Hydrocortisone-conjugated Glycine Protects the Mast Cells Against Compound 48/80 Induced Asthma in Rats

目的:观察甘氨酸偶联皮质醇(HG)对Compound 48/80(C48/80)刺激大鼠急性过敏反应的影响,以期为后期深入探讨糖皮质激素非基因组机制建立有效研究工具。方法:将60只雄性SD大鼠随机分为4组:急性过敏模型组(腹腔注射C48/80+对照溶剂,n=15)、皮质醇(GC)干预组(腹腔注射C48/80+GC,n=15)、HG干预组(腹腔注射C48/80+HG,n=15)、空白对照组(腹腔注射等量生理盐水,n=15)。造模组及各干预组分别在腹腔注射C48/80 5 min后分别给予腹腔注射对照溶剂(等量无水乙醇,为GC及HG的稀释溶剂)、GC或HG,之后每5 min监测大鼠肛温及平均动脉压(MAP),腹腔注射C48/80后30 min时处死大鼠,取大鼠咽喉组织及肺组织,多聚甲醛固定切片后观察组织形态;同时摘眼球取血,分离血清,采用酶联免疫吸附实验(ELISA)测定血清中组胺含量。结果:与空白对照组相比,造模组大鼠肛温及平均动脉压呈持续降低趋势,C48/80刺激可诱发大鼠咽喉及肺组织水肿、肺泡损坏及炎性细胞浸润,大鼠血清中组胺水平较对照组显著升高(P<0.05);较造模组各干预组均可减轻造模诱发的上述过敏反应(P<0.05),但HG干预组较GC干预组促进大鼠肛温降低恢复起效时间快、复温效率高(P<0.05),HG干预组减弱大鼠平均动脉压持续性降低趋势较GC干预组效果明显(P<0.05);HG干预组显著减轻C48/80刺激导致的咽喉及肺组织水肿、肺泡损坏及炎性细胞浸润;HG干预组能显著抑制C48/80刺激导致的血清组胺升高水平(P<0.05),但HG干预组与GC干预组抑制效果无明显差异(P>0.05)。结论:HG能够快速抑制C48/80刺激诱导的大鼠急性过敏反应;能够显著抑制肥大细胞组胺的释放;HG可能通过非基因组作用快速抑制肥大细胞的脱颗粒,抑制炎性介质组胺的释放,从而抑制免疫炎症反应,发挥抗过敏及抗炎症效应,从而为研究糖皮质激素非基因组机制提供了有效的研究工具。

Objective： To observe the effect of Hydrocortisone-conjugated glycine （HG） on allergic reaction induced by Compound 48/80 （C48/80） in rats. To establish an effective research tool for later in-depth study of glucocorticoid nongenomic mechanism. Methods： 60 Male SD rats randomly divided into 4 groups： acute allergic group （intraperitoneal injection C48/80＋vehicle control, n=15）, cortisol （GC）intervention group （intraperitoneal injection C48/80＋GC, n=15）, HG intervention group（intraperitoneal injection C48/80＋HG,n=15） and a normal control group（peritoneal injection with the same amount of normal sodium,n=15）. Module and intervention groups injected intraperitoneally with C48/80 were injected with vehicle control （Equal amount of anhydrous ethanol, diluted solvent for GC and HG）, GC or HG respectively 5 mins later. Rectal temperature and mean arterial pressure were monitored every 5 rains. After intraperitoneal injection of C48/80 30 mins rats were sacrificed, took the throat tissue and lung tissue of rats, paraformaldehyde fixed and sliced to observe with morphology. At the same time rat serums were collected, serum histamine content wasdetermined by ELISA Kit. Results： Compared with the control group, the rectal temperature and mean arterial pressure of the rats in the model group showed a decreasing trend, C48/80 stimulation can induce edema, alveolar damage and inflammatory cell infiltration in rat throat and lung tissue, the histamine level in serum of rats was significantly higher than that of control group （P〈0.05）; Compared with the model group, the above mentioned allergic reactions could be reduced by the intervention groups（P〈0.05）, but the HG intervention group promotes rat rectal temperature reduce recovery, a faster onset time, higher efficiency compared with the GC intervention group （P〈0.05）; HG intervention group significantly reduced the throat and lung edema, alveolar damage and inflammatory cell infiltration induced by C48/80 stimulation; HG intervention group significantly attenuated the average arterial pressure decrease in rats than the GC intervention group（P〈0.05 ）; HG intervention group can significantly inhibit the increase of serum histamine level caused by C48/80 stimulation （P〈0. 05）, however, there was no significant difference between the HG and the GC intervention group on the total level of histamine inhibition in 30min. Conclusions： HG can inhibit acute allergic reaction induced by C48/80 in rats rapidly; Can inhibit the release of histamine in mast cells significantly; HG may inhibit the degranulation of mast cells and inhibit the release of histamine in inflammatory mediators through nongenomic mechanism, thereby inhibiting immune inflammatory response and exert anti-allergic and anti-inflammatory effects, thus provides an effective research tool for studying the nongenomic mechanism of glucocorticoids.