丹参酮IIA对大鼠脊髓损伤后神经保护作用的实验研究

发布时间：2019-06-13 23:15

【摘要】：背景：脊髓损伤导致了一系列严重的并发症,包括瘫痪、疼痛以及进行性的神经功能障碍等。这不仅对患者本人的身心是一个巨大的打击,也为社会增加了沉重的经济负担。基础研究、病例报道和临床试验都表明早期治疗可以改善神经功能的恢复。但迄今为止,还没有成熟的治疗方法能够对神经学的结果产生直接的治疗作用,因此需要我们不断的探索新的治疗手段。近年来很多基础科学、实验研究和临床研究都致力于防止继发性损伤,促进再生。这些研究为我们更好的理解脊髓损伤后复杂病理事件之间的相互联系提供了很好的帮助,也为我们未来的研究指明了方向。目的：探索丹参酮ⅡA对脊髓损伤大鼠后肢神经功能障碍和膀胱功能障碍的影响,并初步探讨可能的作用机制。方法：将64只大鼠随机分为四组：对照组(仅切除椎板,尾静脉注射生理盐水1ml/d,连续7天),模型组(使用NYU Impactor脊髓打击器选择25mm高度造成T9脊髓节段不完全损伤动物模型,术后予尾静脉注射生理盐水1ml/d,连续7天),丹参酮ⅡA组(造模成功后即刻予尾静脉注射丹参酮ⅡA磺酸钠注射液2Omg/Kg/d,连续给药7天),甲强龙组(造模成功后即刻予尾静脉注射甲基强的松龙30mg/Kg)。(1)采用BBB评分法评价各组大鼠后肢运动功能的恢复；(2)通过热板实验评价各组大鼠后肢感觉功能的恢复；(3)在术后7天、4周、8周时进行心脏灌注固定,取材T9脊髓和L6-S1背根神经节；通过HE染色、尼氏染色观察脊髓、L6-S1背根神经节局部组织结构的变化,并使用Ⅰ：mage-Pro Plus 6.0病理分析软件测量背根神经节细胞横截面的面积；(4)术后4周取材膀胱组织,通过Masson染色膀胱壁后观察组织结构的病理变化；(5)术后7天、术后8周通过免疫组化法标记各组脊髓中的巨噬/小胶质细胞；(6)术后4周通过尿动力学评价各组大鼠的膀胱功能。结果：(1)丹参酮ⅡA组和甲强龙组大鼠后肢运动功能都有显著改善,但甲强龙组改善更明显(P0.05)；(2)术后3天,甲强龙组感觉功能恢复较丹参酮ⅡA组快,差异有统计学意义(P0.05)；术后7天,丹参酮ⅡA组和甲强龙组感觉功能恢复程度接近；自术后2周开始,丹参酮ⅡA组表现出了比甲强龙组更明显的改善趋势,但并未达到统计学差异(P0.05)。(3)术后3天,除对照组,余各组部分巨噬/小胶质细胞形态由分支状变为杆状或圆状,其中丹参酮ⅡA组和甲强龙组形态改变的细胞较少,数量较模型组明显减少；(4)术后4周,除对照组,余各组巨噬/小胶质细胞形态大部分转变为分枝状,其中模型组大鼠细胞数量较对照组明显减少,丹参酮ⅡA组细胞数量较模型组显著增加,而甲强龙组细胞数量较模型组增加更为明显。结论：(1)丹参酮IIA能够促进脊髓损伤大鼠后肢运动、感觉功能恢复；(2)丹参酮IIA能够改善脊髓损伤后脊髓、膀胱、神经节的病理进展,促进巨噬/小胶质细胞对神经恢复的有利影响；(3)丹参酮IIA能够提高慢性脊髓损伤大鼠排尿效率,减少无排尿性收缩,促进下尿路功能障碍的恢复；(4)尿动力学是评价大鼠下尿路功能的有效手段。
[Abstract]:Background: Spinal cord injury has resulted in a series of serious complications, including paralysis, pain, and progressive neurological dysfunction. This is not only a great blow to the body and mind of the patient, but also a heavy economic burden for society. Basic studies, case reports, and clinical trials have shown that early treatment can improve the recovery of neurological function. But to date, there is no mature method of treatment that can have a direct therapeutic effect on the neurological outcome, so we need to continue to explore new treatments. In recent years, many basic science, experimental research and clinical research work to prevent secondary injury and promote regeneration. These studies provide a good help for our better understanding of the interconnectedness of complex and pathological events after spinal cord injury, as well as directions for our future studies. Objective: To explore the effect of Tanshinone 鈪 on the neurological and bladder dysfunction in the hindlimb of rats with spinal cord injury, and to explore the possible mechanism of action. Methods: Sixty-four rats were randomly divided into four groups: the control group (only the lamina was cut off, the tail vein was injected with normal saline for 1 ml/ d for 7 days), and the model group (using the NYU Impactor's spinal cord) to select the 25 mm height did not completely damage the animal model of the T9 spinal cord. After the operation,1 ml/ d of the normal saline (1 ml/ d,7 days for 7 days) was given, and the tanshinone 鈪 group (the injection of the tanshinone 鈪 sodium sulfonate injection was 2 Omg/ Kg/ d for 7 days after the model was successfully established), and the group A (30 mg/ Kg of methylprednisolone was given to the tail immediately after the model was successfully established). (1) The recovery of the hind limb movement function of each group was evaluated by the BBB scoring method; (2) the recovery of the sensory function of the hind limb of each group was evaluated by a hot plate experiment; (3) the cardiac perfusion was carried out at 7,4 and 8 weeks after the operation, and the spinal cord and the L6-S1 dorsal root ganglion were obtained. The changes of the local tissue structure of the dorsal root ganglion of the spinal cord and the L6-S1 dorsal root ganglion were observed by HE staining, and the area of the cross-section of the dorsal root ganglion cells was measured by using the I: mage-Pro Plus 6.0 pathology analysis software. (4) The bladder tissue was obtained at 4 weeks after operation. The pathological changes of the tissue structure were observed following the staining of the bladder wall by Masson; (5) the macrophagocytic/ microglial cells in each group of the spinal cord were labeled by immunohistochemistry in 7 days after the operation; and (6) the bladder function of each group was evaluated by urodynamics at 4 weeks after operation. Results: (1) There was a significant improvement in the exercise function of the hind limbs of the group A and the group A of the group A, but the improvement of the group A was more obvious (P0.05); (2) The sensory function of the group A was faster than that of the tanshinone 鈪 group after 3 days of operation (P0.05); and after the operation for 7 days, The degree of recovery of the sensory function of the tanshinone 鈪 group and the group A of the group A was close to that of the group A. From the second week after the operation, the group A of the tanshinone 鈪 showed a more obvious improvement tendency than that of the group A, but did not reach the statistical difference (P0.05). (3) After 3 days of operation, in addition to the control group, the morphology of the macrophagocytic/ microglial cells in each group was changed into a rod shape or a round shape, and the number of cells of the tanshinone 鈪 group and the group A of the group A was less and the number of the group was significantly decreased; and (4)4 weeks after the operation, the control group was removed. In the model group, the number of cells in the model group was significantly decreased, and the number of cells in the group A was significantly increased compared with that of the control group, while the number of cells in the group A was more obvious than that of the model group. Conclusion: (1) Tanshinone IIA can promote the hind limb movement and sensory function recovery of spinal cord injury; (2) Tanshinone IIA can improve the pathological progress of spinal cord, bladder and ganglion after spinal cord injury, and promote the beneficial effect of macrophagocyte/ microglial cell on nerve recovery; (3) Tanshinone IIA can improve the micturition efficiency of rats with chronic spinal cord injury, reduce the contractility and promote the recovery of lower urinary tract dysfunction; and (4) the urodynamics is an effective means to evaluate the function of lower urinary tract in rats.
【学位授予单位】：北京中医药大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R651.2

【相似文献】