Peng Lingling, Wang Jingbo, Pan Yuzheng, Du Yiheng, Huang Guihua
Guangxi University of Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China.
Department of Traditional Chinese Medicine, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China. Corresponding author: Huang Guihua, Email:
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2021 Nov;33(11):1347-1352. doi: 10.3760/cma.j.cn121430-20210709-01030.
To observe the effects of the Chinese medicine prescription Xiao-Cheng-Qi decoction (XCQD) on acute brain edema and inflammatory factors in rats with severe traumatic brain injury (sTBI).
A total of 108 male Sprague-Dawley (SD) rats were divided into control group, sham operation group, sTBI model group, and XCQD low, medium, high dose groups by random number table method, with 18 rats in each group. sTBI rat model was prepared according to the modified Freeney method. At 6 hours after injury, the XCQD low, medium, and high dose groups were given XCQD 1.80, 2.78, and 4.59 g/kg by gavage, respectively, and the other three groups were given the same amount of normal saline, once a day for 3 days. After 3 days of injury, rats in each group were sacrificed after the modified neurologic severity score (mNSS) assessed. Pathological changes of brain tissue were observed under light microscope after hematoxylin eosin (HE) staining, water content of brain tissue was measured by dry-wet specific gravity method, and the expressions of aquaporin 4 (AQP4), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in brain tissue were detected by Western blotting. Serum TNF-α and IL-1β levels were detected by enzyme linked immunosorbent assay (ELISA).
Compared with the normal group, the mNSS score of rats increased significantly, the structure of brain tissue was disordered, and pathological changes appeared such as inflammation, edema, pyknosis of nerve nuclei, water content, the protein expressions of AQP4, TNF-α and IL-1β in brain tissue, and the contents of TNF-α, IL-1β in serum were significantly increased. After XCQD intervention, the above indexes were significantly improved. Compared with sTBI model group, the mNSS score of XCQD medium and high dose groups significantly decreased (6.94±1.16, 6.88±1.02 vs. 8.61±1.09, both P < 0.05), and the pathological changes such as brain edema and inflammation were alleviated. Brain tissue water content, AQP4 protein expression and contents of serum TNF-α, IL-1β in XCQD low, medium, and high dose groups significantly decreased compared with sTBI model group [brain tissue water content: (78.25±0.71)%, (77.62±0.44)%, (76.70±0.74)% vs. (80.08±0.66)%; the expression of brain AQP4 protein (AQP4/β-actin): 0.86±0.13, 0.84±0.22, 0.65±0.13 vs. 1.08±0.14; serum TNF-α (ng/L): 106.34±15.07, 95.75±17.26, 89.00±17.36 vs. 141.96±29.47; serum IL-1β (ng/L): 90.41±12.88, 72.82±13.51, 71.32±16.79 vs. 128.57±22.56, respectively, all P < 0.05]. The protein expressions of TNF-α,IL-1β in brain tissue of XCQD medium and high dose groups also significantly decreased compared with sTBI model group [TNF-α (TNF-α/β-actin): 0.90±0.24, 0.79±0.35 vs. 1.17±0.15; IL-1β (IL-1β/β-actin): 0.91±0.21, 0.68±0.28 vs. 1.23±0.08, respectively, all P < 0.05]. Brain tissue water content, the expression of brain AQP4 protein, the levels of brain tissue and serum IL-1β in XCQD high dose group improved more significant than those of XCQD low dose group.
XCQD can alleviate the acute brain edema in sTBI rats, and it is dose-dependent. The mechanism may be relevant to reduce the secondary inflammatory response of sTBI by inhibiting the expression of inflammatory factors TNF-α and IL-1β.
观察中药方剂小承气汤(XCQD)对重型颅脑损伤(sTBI)大鼠急性脑水肿及炎症因子的影响。
将108只雄性Sprague-Dawley(SD)大鼠按随机数字表法分为对照组、假手术组、sTBI模型组、XCQD低、中、高剂量组,每组18只。参照改良的Feeney法制备sTBI大鼠模型。伤后6小时,XCQD低、中、高剂量组分别按1.80、2.78、4.59 g/kg灌胃给予XCQD,其余三组给予等量生理盐水,每日1次,连续3天。伤后3天,评估改良神经功能缺损评分(mNSS)后处死各组大鼠。苏木精-伊红(HE)染色后在光学显微镜下观察脑组织病理变化,采用干湿比重法测量脑组织含水量,通过蛋白质免疫印迹法检测脑组织中水通道蛋白4(AQP4)、肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)的表达。采用酶联免疫吸附测定(ELISA)法检测血清TNF-α和IL-1β水平。
与正常组相比,大鼠mNSS评分显著升高,脑组织结构紊乱,出现炎症、水肿、神经细胞核固缩等病理变化,脑组织含水量、脑组织中AQP4、TNF-α和IL-1β的蛋白表达以及血清中TNF-α、IL-1β含量均显著增加。XCQD干预后,上述指标均显著改善。与sTBI模型组相比,XCQD中、高剂量组mNSS评分显著降低(6.94±1.16,6.88±1.02比8.61±1.09,P均<0.05),脑水肿和炎症等病理变化减轻。XCQD低、中、高剂量组脑组织含水量、AQP4蛋白表达及血清TNF-α、IL-1β含量与sTBI模型组相比均显著降低[脑组织含水量:(78.25±0.71)%,(77.62±0.44)%,(76.70±0.74)%比(80.08±0.66)%;脑AQP4蛋白表达(AQP4/β-肌动蛋白):0.86±0.13,0.84±0.22,0.65±0.13比1.08±0.14;血清TNF-α(ng/L):106.34±15.07,95.75±17.26,89.00±17.36比141.96±29.47;血清IL-1β(ng/L):90.41±12.88,72.82±13.51,71.32±16.79比128.57±22.56,P均<0.05]。XCQD中、高剂量组脑组织中TNF-α、IL-1β蛋白表达与sTBI模型组相比也显著降低[TNF-α(TNF-α/β-肌动蛋白):0.90±0.24,0.79±0.35比1.17±0.15;IL-1β(IL-1β/β-肌动蛋白):0.91±0.21,0.68±0.28比1.23±0.08,P均<0.05]。XCQD高剂量组脑组织含水量、脑AQP4蛋白表达、脑组织及血清IL-1β水平改善程度均较XCQD低剂量组更显著。
XCQD可减轻sTBI大鼠急性脑水肿,且呈剂量依赖性。其机制可能与抑制炎症因子TNF-α和IL-1β表达从而减轻sTBI的继发性炎症反应有关。