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肾上腺素、瞬时受体电位香草酸1通道和P物质在急性脊髓损伤大鼠并发肺损伤中的变化及意义
喻林,储瑞亮,毕杨,何波,南国新*
0
(重庆医科大学附属儿童医院骨二科, 儿童发育疾病研究教育部重点实验室, 儿童发育重大疾病国家国际科技合作基地, 儿科学重庆市重点实验室, 重庆市干细胞治疗工程技术研究中心, 重庆 400014
*通信作者)
摘要:
目的 探讨肾上腺素、瞬时受体电位香草酸1(TRPV1)通道和P物质在大鼠急性脊髓损伤(ASCI)并发肺损伤中的变化及意义。方法 将228只雌性SD大鼠按随机数字表法分为假手术组(90只)、ASCI组(108只)、双侧肾上腺切除组(15只)、双侧肾上腺切除后ASCI组(15只)。采用改良Allen's打击模型(打击锤质量为10 g,打击高度为25 mm)于T10脊髓节段制备ASCI模型,假手术组仅暴露T10节段脊髓,双侧肾上腺切除后ASCI组于肾上腺切除术5 d后制作ASCI模型。采用高效液相色谱法检查大鼠血清肾上腺素水平。取大鼠肺组织标本,计算肺湿干质量比以反映肺组织水肿变化,采用H-E染色检测肺组织病理学变化,采用免疫组织化学染色、蛋白质印迹法检测肺组织中TRPV1蛋白表达,采用酶联免疫吸附试验检测肺组织中P物质的含量。结果 脊髓损伤后2、6、12、24、48、72 h,ASCI组大鼠血清肾上腺素水平均高于假手术组,差异均有统计学意义(P均<0.01)。脊髓损伤后24、48、72 h,ASCI组大鼠肺水肿和肺损伤组织病理学变化逐渐加重,损伤1周时开始恢复。脊髓损伤后24、48、72 h,ASCI组大鼠肺组织TRPV1蛋白表达量和P物质含量均较假手术组升高,差异均有统计学意义(P均<0.05,P均<0.01)。脊髓损伤后72 h,双侧肾上腺切除后ASCI组肺组织水肿和组织病理学变化均较ASCI组减轻。结论 肾上腺素可能参与大鼠ASCI并发的肺水肿和肺损伤进程,这种效应可能与肺组织中TRPV1、P物质的表达上调有关;双侧肾上腺切除预处理可减轻ASCI并发的肺水肿和肺损伤程度。
关键词:  脊髓损伤  肺损伤  肾上腺素  瞬时受体电位香草酸1通道  P物质
DOI:10.16781/j.0258-879x.2019.08.0866
投稿时间:2019-05-07修订日期:2019-07-10
基金项目:国家自然科学基金(81272172).
Adrenaline, transient receptor potential vanilloid 1 channel and substance P in rats with acute spinal cord injury complicated with lung injury
YU Lin,CHU Rui-liang,BI Yang,HE Bo,NAN Guo-xin*
(Department of Orthopedics(Ⅱ), Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing Key Laboratory of Pediatrics, Chongqing Engineering Research Center of Stem Cell Therapy, Chongqing 400014, China
*Corresponding author)
Abstract:
Objective To study the changes and significance of adrenaline, transient receptor potential vanilloid 1 (TRPV1) channel and substance P in rats with acute spinal cord injury (ASCI) complicated with lung injury. Methods Two hundred and twenty eight female Sprague-Dawley rats were randomly divided into sham operation group (sham group, n=90), ASCI group (n=108), bilateral adrenalectomy group (n=15), and ASCI after bilateral adrenalectomy group (n=15). The ASCI model was established on the T10 spinal cord segment using a modified Allen's strike model (10 g, 25 mm). The sham group only exposed the T10 spinal cord, and the ASCI after bilateral adrenalectomy group was established 5 days after bilateral adrenalectomy. High-performance liquid chromatography was used to detect the changes of serum adrenaline. The pulmonary tissue specimens were collected from rats. Wet-to-dry lung weight ratio was used to detect the changes of pulmonary edema, and H-E staining was used to detect the pathological changes of lung tissue. The expression of TRPV1 protein in lung tissue was detected by immunohistochemistry and Western blotting. The contents of substance P in the lung tissue were detected by enzyme-linked immunosorbent assay. Results Serum adrenaline levels in the ASCI group were significantly higher than those in the sham group at 2, 6, 12, 24, 48, and 72 h after spinal injury (all P<0.01). The pulmonary edema and lung injury gradually aggravated in the ASCI group at 24, 48 and 72 h after spinal injury, and began to recover at 1 week. The expression levels of TRPV1 protein and substance P contents in ASCI group were significantly upregulated compared with the sham group at 24, 48, and 72 h after spinal injury (all P<0.05, P<0.01). The edema of lung tissue and pathological injury in the ASCI after bilateral adrenalectomy group were alleviated compared with the ASCI group 72 h after spinal injury. Conclusion Adrenaline may involve in the pathogenesis of pulmonary edema and lung injury in rats with ASCI, which may be related to the upregulation of TRPV1 and P substance expression. The pulmonary edema and lung injury after ASCI can be alleviated by adrenalectomy.
Key words:  spinal cord injuries  lung injuries  adrenaline  transient receptor potential vanilloid 1 channel  substance P