孕酮调节创伤性脑损伤后炎症反应及预后的实验研究

目的探索孕酮在创伤性脑损伤(TBI)中是否能调节炎症反应及改善神经功能预后。方法将小鼠随机分为假手术组、孕酮治疗组和对照组。构建TBI模型后,孕酮治疗组给予孕酮治疗,对照组予以等量生理盐水。用mNSS评分检测小鼠神经功能,干湿重法检测脑组织含水量,蛋白印迹及免疫组化法检测脑组织中炎性因子及炎性细胞含量,流式细胞术检测脾脏中调节性T细胞(Treg)的数量。结果我们的实验研究发现孕酮治疗组小鼠mNSS评分在脑创伤后第3、5、7天时明显低于对照组(P0.05);孕酮治疗组脑组织含水量在伤后第1、3、5、7天较对照组相比明显减少(P0.05)。在伤后第1、3天时,与假手术组相比,孕酮治疗组和对照组炎性因子(IL-1β、TNF-α、NF-κB、IL-10)和炎性细胞(GFAP、Iba-1)含量显著增加(P0.05);与对照组相比,孕酮治疗组炎性因子和炎性细胞含量明显下降(P0.05),且脾脏内Treg数量显著增多(P0.05)。在伤后第3天,孕酮治疗组及假手术组脑损伤灶处CD3及MPO细胞数量无显著性差异,均明显低于对照组(P0.05)。结论孕酮治疗可以改善小鼠TBI后的神经功能预后,其机制可能与孕酮在脑创伤后的免疫炎症调节作用有关。

Studies on the role of progesterone in the regulation of immune inflammation and the improvement of prognosis in traumatic brain injury

［Abstract］ Objective To explore whether progesterone regulates inflammatory and improves neurological outcomes in traumatic brain injury. Methods The mice were randomly assigned into three groups： sham-operated group， PROG-treated group and traumatic control group. The TBI model was done by the controlled contusion instrument. Progesterone treatment （16 mg/kg） was initiated intraperitoneally after injury with PROG-treated group， and the control group was injected with normal saline with the same dose. Neurological function was evaluated by the Modified Neurological Severity Score （mNSS）。 Dry-wet weight method was conducted in order to determine the water content in the brain tissue. The Western Blot was underwent to assess inflammatory cytokines like IL-10， IL-1β， TNF-α， NF-κB. The number of inflammatory cells （MPO， CD3， GFAP and Iba-1） cells were detected by immunocytochemistry. Quantitive determination of Treg cells in spleen was done by flow cytometry. Results Our study found that： the mNSS score of PROG-treated group are significantly lower than control group after TBI third， fifth， seventh day （P<0.05）. The water content in the brain tissue of PROG-treated group are obviously lower than control group after TBI fist， third， fifth， seventh day （P<0.05）. Compared to the sham-operated group， inflammatory cytokines， like IL-10， IL-1β， TNF-α， NF-κB， and inflammatory cells （MPO， CD3， GFAP and Iba-1） in the PROG-treated group and the control group was discovered more （P<0.05）. Compared to the control group ， inflammatory cytokines inflammatory cells in the PROG-treated group was very fewer （P<0.05）， and the Treg cells in spleen was significant increased. The PROG-treated group and the sham operation group has no difference in the number of MPO and CD3 after TBI three days， but all lower than the control group. Conclusion Progesterone treatment can improve the neurological outcome after TBI， and its mechanism may be related to the regulation of progesterone on immune inflammation after brain injury.