目的 探讨大黄素对大鼠急性脊髓损伤后脊髓水肿的影响及其机制。方法 将180只健康雌性Sprague-Dawley大鼠随机分为假手术组(A组),模型组(B组),大黄素低剂量组(C组)、中剂量组(D组)和高剂量组(E组),每组36只。采用Allen法制作大鼠脊髓损伤模型,术后3 d、7 d、14 d及28 d,BBB评分及斜板实验观察大鼠运动功能恢复。术后3 d,HE染色观察脊髓组织病理变化,干湿重法检测脊髓组织含水量,伊文思蓝(EB)染色检测血脊髓屏障(BSCB)通透性,RT-PCR和Western blotting检测水通道蛋白-4 (AQP-4)、基质金属蛋白酶-2 (MMP-2) mRNA和蛋白表达。结果 BBB评分及斜板实验结果显示,术后7 d、14 d、28 d,C组、D组和E组均优于B组(P<0.05),E组最佳(P<0.05)。术后3 d,HE染色显示,B组损伤节段内有大片出血灶,神经细胞肿胀、破坏,大量炎性细胞浸润,组织间隙增宽,水肿严重,C组、D组和E组上述病理改变改善,其中E组最明显;B组脊髓组织含水量较A组增高(P<0.05),D组、E组脊髓组织含水量低于B组和C组(P<0.05),E组低于D组(P<0.05);B组EB含量高于A组(P<0.05),C组、D组和E组EB含量均低于B组(P<0.05);C组、D组和E组AQP-4、MMP-2 mRNA和蛋白表达均低于B组(P<0.05),E组最低(P<0.05)。结论 大黄素能减轻大鼠急性脊髓损伤后的脊髓水肿,促进肢体功能康复,这可能与大黄素下调损伤后脊髓组织AQP-4和MMP-2表达,降低BSCB通透性有关。
Objective To investigate the effect of emodin on spinal cord edema induced by acute spinal cord injury (SCI) and its mechanism.Methods A total of 180 healthy female Sprague-Dawley rats were randomly divided into sham group (group A), model group (group B), and low-dose group (group C), middle-dose group (group D) and high-dose group (group E) of emodin, with 36 cases in each group. The SCI model was established with the modified Allen's method. Functional recovery was evaluated with Basso-Beattie-Bresnahan (BBB) score and inclined plate test three days, seven days, 14 days and 28 days after modeling. Three days after modeling, the pathological changes of the spinal cord were observed by HE staining; the water content of spinal cord was detected by dry-wet weight method, the blood-spinal cord barrier (BSCB) permeability was detected by Evans blue (EB) staining, and the expression of aquaporin-4 (AQP-4) and matrix metalloproteinase-2 (MMP-2) mRNA and protein were detected by RT-PCR and Western blotting respectively. Results The BBB score and inclined plate scores were better in groups C, D and E than in group B (P<0.05) seven days, 14 days and 28 days after modeling, especially in group E (P<0.05). Three days after modeling, HE staining showed that there was a large hemorrhage in the section of group B, the nerve cells were swollen and damaged, and a large number of inflammatory cells were infiltrated, the tissue gap was widened and the edema was severe. The above pathological changes were better in groups C, D and E than in group B, especially in group E. The spinal cord water content was higher in group B than in group A (P<0.05), and was lower in groups D and E than in groups B and C (P<0.05). EB content was higher in group B than in group A (P<0.05), and was lower in groups C, D and E than in group B (P<0.05). The expression of AQP-4, MMP-2 mRNA and protein were lower in groups C, D and E than in group B (P<0.05), especially in group E (P<0.05). Conclusion Emodin can alleviate spinal cord edema, and improve hind limb movement function after SCI, which could be related with the down-regulation of AQP-4 and MMP-2 expression, and the reduction of the permeability of BSCB.
[1] 朱彦东,周开升,郭永强,等. 大鼠脊髓损伤后ski相关蛋白表达的时空变化规律及作用[J]. 中国康复理论与实践, 2017, 23(8): 912-918.
[2] Oyihbo CA.Secondary injury mechanisms in traumatic spinal cord injury: a nugget of this multiply cascade[J]. Act Neurobiol Exp, 2011, 71(2): 281-299.
[3] 刘理静,钱红,张平. 大黄素对肺纤维化大鼠的保护作用及部分机制研究[J]. 中国药理学通报, 2015, 31(2): 266-272.
[4] 陈素领,周杰超,张杰,等. 大黄素通过抑制FOXO1活性减轻NO对神经细胞的损伤[J]. 生物化学与生物物理进展, 2016, 43(11): 1076-1085.
[5] Sun Y, Sun L, Liu S, et al.Effect of emodin on aquaporin 5 expression in rats with sepsis-induced acute lung injury[J]. J Tradit Chin Med, 2015, 35(6): 679-684.
[6] Xu J, Huang B, Wang Y, et al.Emodin ameliorates acute lung injury induced by severe acute pancreatitis through the up-regulated expressions of AQP1 and AQP5 in lung[J]. Clin Exp Pharmacol, 2016, 43(11): 1071-1079.
[7] Zhao XY, Qiao GF, Li BX, et al.Hypoglycaemic and hypolipidaemi effects of emodin and its effect on L-type calcium channels in dyslipidaemic diabetic rats[J]. Clin Exp Pharmacol, 2009, 36(1): 29-34.
[8] 郑利强,伍亚民,石永江,等. 黄芪多糖对大鼠脊髓损伤后运动功能和脊髓病理的效果[J]. 中国康复理论与实践, 2016, 22(11): 1269-1275.
[9] 闵友江,程立红,姚海华,等. “三通针法”对脊髓损伤大鼠p75神经营养素受体表达的影响[J]. 中国康复理论与实践, 2017, 23(6): 621-627.
[10] He Z, Zou S, Yin J, et al.Inhibition of endoplasmic reticulum stress preserves the integrity of blood-spinal cord barrier in diabetic rats subjected to spinal cord injury[J]. Sci Rep, 2017, 7(1): 7661-7675.
[11] Pei J, Fan L, Nan K, et al.HSYA alleviates secondary neuronal death through attenuating oxidative stress, inflammatory response, and neural apoptosis in SD rat spinal cord compression injury[J]. J Neuroinflamm, 2017, 14(1): 97-111.
[12] Yan X, Liu J, Wang X, et al.Pretreatment with AQP4 and NKCC1 inhibitors concurrently attenuated spinal cord edema and tissue damage after spinal cord injury in rats[J]. Front Physiol, 2018, 9: 6.
[13] Li XQ, Chen FS, Tan WF, et al.Elevated microRNA-129-5p level ameliorates neuro inflammation and blood-spinal cord barrier damage after ischemia-reperfusion by inhibiting HMGB1 and the TLR3-cytokine pathway[J]. J Neuroinflamm, 2017, 14(1): 205-217.
[14] Li XQ, Lv HW, Tan WF, et al.Role of the TLR4 pathway in blood-spinal cord barrier dysfunction during the bimodal stage after ischemia/reperfusion injury in rats[J]. J Neuroinflamm, 2014, 11(1): 62-73.
[15] Bi Y, Zhu Y, Zhang M, et al.Effect of shikonin on spinal cord injury in rats via regulation of HMGB1/TLR4/NF-kB signaling pathway[J]. Cell Physiol Biochem, 2017, 43(2): 481-491.
[16] 张晨,吕雷锋,李苗,等. 藏红花通过抗凋亡、抗炎和抗水肿机制发挥对大鼠脊髓损伤模型的神经保护作用[J]. 西安交通大学学报(医学版), 2017, 38(2): 280-289.
[17] 熊春翔,宗少晖,曾高峰,等. 大鼠Allen's脊髓损伤模型的建立及评价[J]. 广西医科大学学报, 2011, 28(2): 215-217.
[18] 周华,刘华,黄坚. 基质金属蛋白酶-9与脊髓损伤后脊髓水肿的关系[J]. 中国脊柱脊髓杂志, 2007, 17(1): 59-61, 83.
[19] 胥少汀,郭世绂. 脊髓损伤的基础与临床[M]. 2版.北京:人民卫生出版社, 2002: 220-222.
[20] Cui M, Ma X, Sun J, et al.Effects of STAT3 inhibitors on neural functional recovery after spinal cord injury in rats[J]. Biosci Trends, 2016, 10(6): 460-466.
[21] Zhang D, Xuan J, Zheng B, et al.Metformin improves functional recovery after spinal cord injury via autophagy flux stimulation[J]. Mol Neurobiol, 2017, 54(5): 3327-3341.
[22] Hu AM, Li JJ, Sun W, et al.Myelotomy reduces spinal cord edema and inhibits aquaporin-4 and aquaporin-9 expression in rats with spinal cord injury[J]. Spinal Cord, 2015, 53(2): 98-102.
[23] Liu X, Wang Y, Yang J, et al.Anti-edema effect of melatonin on spinal cord injury in rats[J]. Biomed Pap, 2015, 159(2): 220-226.
[24] Patel NP, Huang JH.Hyperbaric oxygen therapy of spinal cord injury[J]. Med Gas Res, 2017, 7(2): 133-143.
[25] Kumar H, Ropper AE, Lee SH, et al.Propitious therapeutic modulators to prevent blood-spinal cord barrier disruption in spinal cord injury[J]. Mol Neurobiol, 2017, 54(5): 3578-3590.
[26] Miranpuri GS, Schomberg DT, Alrfaei B, et al.Role of matrix metalloproteinases 2 in spinal cord injury-induced neuropathic pain[J]. Ann Neurosci, 2016, 23(1): 25-32.
