Article(id=1208154041171947888, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1208154038609228128, articleNumber=null, orderNo=null, doi=10.11855/j.issn.0577-7402.2022.02.0136, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1625673600000, receivedDateStr=2021-07-08, revisedDate=null, revisedDateStr=null, acceptedDate=1630857600000, acceptedDateStr=2021-09-06, onlineDate=1765975969831, onlineDateStr=2025-12-17, pubDate=1645977600000, pubDateStr=2022-02-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1765975969831, onlineIssueDateStr=2025-12-17, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1765975969831, creator=13701087609, updateTime=1765975969831, updator=13701087609, issue=Issue{id=1208154038609228128, tenantId=1146029695717560320, journalId=1189873630562394117, year='2022', volume='47', issue='2', pageStart='107', pageEnd='212', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1765975969218, creator=13701087609, updateTime=1765976148463, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1208154790459192257, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1208154038609228128, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1208154790459192258, tenantId=1146029695717560320, journalId=1189873630562394117, issueId=1208154038609228128, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=136, endPage=142, ext={EN=ArticleExt(id=1208154041826259322, articleId=1208154041171947888, tenantId=1146029695717560320, journalId=1189873630562394117, language=EN, title=Mechanism of cervical skeletal muscle injury induced by +Gz exposure in rabbits, columnId=1190310110212751762, journalTitle=Medical Journal of Chinese People’s Liberation Army, columnName=Basic Research, runingTitle=null, highlight=null, articleAbstract=

Objective To study the mechanism of +Gz environment exposure induced neck muscle injury in rabbits. Methods A total of 20 male rabbits were randomized into the negative control group, +Gz exposure group, +Gz exposure plus neck loading group, and +Gz exposure plus neck loading plus fixation group (5 in each group). The animal centrifuge was used to simulate the +Gz exposure environment. Rabbits in the +Gz exposure group, +Gz exposure plus neck loading group, and +Gz exposure plus neck loading plus fixation group were exposed for 4 weeks in a +6 Gz environment. Before and after the experiment, the rabbits in each group were photographed by X-ray. At the time of 0-, 1-, 2-, 3- and 4-week exposure, ear vein blood was collected from rabbits in each group, and ELISA was performed to detected plasma leveles of lactate dehydrogenase (LDH), creatine kinase (CK) and reactive oxygen species (ROS). After exposure, the trapezius muscle of rabbits was taken for HE staining to detect the muscle morphology.The apoptosis level was detected by TUNEL in the paraffin section of the rabbit trapezius muscle. The expressions of apoptosis-related proteins Bax and Bcl-2 in the trapezius muscle of rabbits were detected by Western blotting. The rabbit's neck trapezius muscle was taken to conduct proteomics testing, select differentially expressed proteins, and perform protein function cluster analysis. Results There was no significant difference in the morphology of the cervical vertebra in each group before and after+Gz exposure. Compared with negative control group, plasma levels of LDH, CK, and ROS in +Gz exposure group, +Gz exposure plus neck loading group, and +Gz exposure plus neck loading plus fixation group significantly increased (P<0.05). Compared with negative control group, the ratio of apoptosis protein Bax/Bcl-2 in +Gz exposure group significantly increased (P<0.05), the number of positive apoptosis cells detected by TUNEL significantly increased (P<0.05). Compared with +Gz exposure group, the+Gz exposure plus neck loading group showed an increased Bax/Bcl-2 ratio (P<0.05) and more TUNEL positive cells (P<0.05).Compared with +Gz exposure plus neck loading group, the ratio of apoptosis protein Bax/Bcl-2 and the positive number of apoptosis were decreased in +Gz exposure plus neck loading plus fixation group, but there were no statistically significant differences (P>0.05).A total of 600 proteins were significantly altered in +Gz exposure group compared to negative control group, and GO functional clustering analysis of the differential proteins revealed significant enrichment of proteins associated with mitochondrial function. Conclusion Exposure to the +Gz environment can cause neck muscle injury and cell apoptosis in rabbits. The muscle injury symptoms can be improved by fixing the stress link of the rabbit neck. Cell apoptosis induced by +Gz exposure may be related to mitochondrial function.

, correspAuthors=Yong-Chun Wang, authorNote=null, correspAuthorsNote=
*E-mail:
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目的 探讨+Gz暴露诱发家兔颈部肌肉损伤的机制。方法 雄性家兔20只,随机分为正常对照组、+Gz暴露组、+Gz暴露+颈部负重组、+Gz暴露+颈部负重+固定组,每组5只。使用动物离心机模拟+Gz暴露环境,+Gz暴露组、+Gz暴露+颈部负重组、+Gz暴露+颈部负重+固定组在+6 Gz环境下连续暴露4周。暴露前和暴露结束后,对各组家兔进行X线摄片;暴露前及暴露1、2、3和4周,采集各组家兔耳缘静脉血,采用ELISA检测血浆乳酸脱氢酶(LDH)、肌酸激酶(CK)、活性氧(ROS)水平;暴露结束后取家兔颈部斜方肌,采用HE染色观察肌肉形态变化,TUNEL检测细胞凋亡情况,Western blotting检测凋亡相关蛋白Bax、Bcl-2的表达情况;取家兔颈部斜方肌行蛋白组学检测,挑选差异表达蛋白并进行GO功能聚类分析。结果 暴露前后各组家兔颈椎形态无明显差异;与正常对照组比较,+Gz暴露组、+Gz暴露+颈部负重组、+Gz暴露+颈部负重+固定组血浆LDH、CK、ROS水平升高(P<0.05);与正常对照组比较,+Gz暴露组凋亡相关蛋白Bax/Bcl-2比值明显增高(P<0.05),TUNEL阳性细胞数明显增多(P<0.05)。与+Gz暴露组比较,+Gz暴露+颈部负重组凋亡相关蛋白Bax/Bcl-2比值明显增高(P<0.05),TUNEL阳性细胞数明显增多(P<0.05);与+Gz暴露+颈部负重组比较,+Gz暴露+颈部负重+固定组凋亡相关蛋白Bax/Bcl-2比值及TUNEL阳性细胞数有所下降,但差异无统计学意义(P>0.05);与正常对照组比较,+Gz暴露组中共600个蛋白发生显著变化,对差异蛋白进行GO功能聚类分析发现其显著富集于线粒体功能相关蛋白。结论 +Gz环境暴露可引起家兔颈部肌肉损伤,导致细胞凋亡,而固定家兔颈部受力环节可缓解肌肉损伤症状;+Gz环境暴露导致的细胞凋亡可能与线粒体功能相关。

, correspAuthors=王永春, authorNote=null, correspAuthorsNote=
王永春,E-mail:
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李卓,硕士研究生,主要从事特殊环境生理学、运动训练与健康促进方面的研究

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李卓,硕士研究生,主要从事特殊环境生理学、运动训练与健康促进方面的研究

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李卓,硕士研究生,主要从事特殊环境生理学、运动训练与健康促进方面的研究

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LDH. 乳酸脱氢酶;CK. 肌酸激酶;ROS. 活性氧;与正常对照组比较,(1)P<0.05

, figureFileSmall=kdjWXQMgro7nGr0UCQJXnQ==, figureFileBig=gZYjv5oxESIqOddqdiri3Q==, tableContent=null), ArticleFig(id=1208154049292120850, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1208154041171947888, language=EN, label=Fig.4, caption=Effect of +Gz exposure on the morphology of skeletal muscle fibers in the neck of rabbits (HE staining), figureFileSmall=D6jImGA0G2XtqoNCDfWXJg==, figureFileBig=xUClDRQPIRb8V+x560fGrg==, tableContent=null), ArticleFig(id=1208154049367618324, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1208154041171947888, language=CN, label=图4, caption=+Gz暴露对家兔颈部骨骼肌纤维形态的影响(HE染色)

A. 正常对照组;B. +Gz暴露组;C. +Gz暴露+颈部负重组;D. +Gz暴露+颈部负重+固定组

, figureFileSmall=D6jImGA0G2XtqoNCDfWXJg==, figureFileBig=xUClDRQPIRb8V+x560fGrg==, tableContent=null), ArticleFig(id=1208154049426338584, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1208154041171947888, language=EN, label=Fig.5, caption=Effect of +Gz exposure on apoptosis of neck muscle cells of rabbits, figureFileSmall=T6B1ek9xd78u/jQ/G4JV9g==, figureFileBig=LqiU9sKqcAwB3nNDGtwo2w==, tableContent=null), ArticleFig(id=1208154049493447451, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1208154041171947888, language=CN, label=图5, caption=+Gz暴露对家兔斜方肌细胞凋亡的影响

A. TUNEL染色检测斜方肌细胞凋亡情况,红色示凋亡细胞,蓝色示细胞核,绿色示骨骼肌肌纤维;B. Western blotting检测家兔斜方肌细胞凋亡相关蛋白Bax、Bcl-2的表达;与正常对照组比较,(1)P<0.05;与+Gz暴露组比较,(2)P<0.05

, figureFileSmall=T6B1ek9xd78u/jQ/G4JV9g==, figureFileBig=LqiU9sKqcAwB3nNDGtwo2w==, tableContent=null), ArticleFig(id=1208154049577333536, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1208154041171947888, language=EN, label=Fig.6, caption=Proteomic analysis of rabbit trapezius muscle, figureFileSmall=0E+Vr7RLrX8s3z8cdeOKXQ==, figureFileBig=yoRHZ6zxzXAsYSYJDH9JRQ==, tableContent=null), ArticleFig(id=1208154049677996833, tenantId=1146029695717560320, journalId=1189873630562394117, articleId=1208154041171947888, language=CN, label=图6, caption=家兔斜方肌蛋白组学分析

A. 家兔斜方肌差异蛋白表达情况;B. 家兔斜方肌差异蛋白GO分析

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+Gz暴露诱发家兔颈部骨骼肌损伤的机制研究
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李卓 , 李曦 , 潘益凯 , 李程飞 , 范洁怡 , 铁娅滕 , 赵星成 , 王永春 *
解放军医学杂志 | 基础研究 2022,47(2): 136-142
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解放军医学杂志 | 基础研究 2022, 47(2): 136-142
+Gz暴露诱发家兔颈部骨骼肌损伤的机制研究
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李卓, 李曦, 潘益凯, 李程飞, 范洁怡, 铁娅滕, 赵星成, 王永春*
作者信息
  • 空军军医大学航空航天医学系航空航天医学训练教研室,西安 710032
  • 李卓,硕士研究生,主要从事特殊环境生理学、运动训练与健康促进方面的研究

通讯作者:

王永春,E-mail:
Mechanism of cervical skeletal muscle injury induced by +Gz exposure in rabbits
Zhuo Li, Xi Li, Yi-Kai Pan, Cheng-Fei Li, Jie-Yi Fan, Ya-Teng Tie, Xing-Cheng Zhao, Yong-Chun Wang*
Affiliations
  • Department of Aerospace Medical Training, School of Aerospace Medicine, Air Force Medical University, Xi’an 710032, China
出版时间: 2022-02-28 doi: 10.11855/j.issn.0577-7402.2022.02.0136
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目的 探讨+Gz暴露诱发家兔颈部肌肉损伤的机制。方法 雄性家兔20只,随机分为正常对照组、+Gz暴露组、+Gz暴露+颈部负重组、+Gz暴露+颈部负重+固定组,每组5只。使用动物离心机模拟+Gz暴露环境,+Gz暴露组、+Gz暴露+颈部负重组、+Gz暴露+颈部负重+固定组在+6 Gz环境下连续暴露4周。暴露前和暴露结束后,对各组家兔进行X线摄片;暴露前及暴露1、2、3和4周,采集各组家兔耳缘静脉血,采用ELISA检测血浆乳酸脱氢酶(LDH)、肌酸激酶(CK)、活性氧(ROS)水平;暴露结束后取家兔颈部斜方肌,采用HE染色观察肌肉形态变化,TUNEL检测细胞凋亡情况,Western blotting检测凋亡相关蛋白Bax、Bcl-2的表达情况;取家兔颈部斜方肌行蛋白组学检测,挑选差异表达蛋白并进行GO功能聚类分析。结果 暴露前后各组家兔颈椎形态无明显差异;与正常对照组比较,+Gz暴露组、+Gz暴露+颈部负重组、+Gz暴露+颈部负重+固定组血浆LDH、CK、ROS水平升高(P<0.05);与正常对照组比较,+Gz暴露组凋亡相关蛋白Bax/Bcl-2比值明显增高(P<0.05),TUNEL阳性细胞数明显增多(P<0.05)。与+Gz暴露组比较,+Gz暴露+颈部负重组凋亡相关蛋白Bax/Bcl-2比值明显增高(P<0.05),TUNEL阳性细胞数明显增多(P<0.05);与+Gz暴露+颈部负重组比较,+Gz暴露+颈部负重+固定组凋亡相关蛋白Bax/Bcl-2比值及TUNEL阳性细胞数有所下降,但差异无统计学意义(P>0.05);与正常对照组比较,+Gz暴露组中共600个蛋白发生显著变化,对差异蛋白进行GO功能聚类分析发现其显著富集于线粒体功能相关蛋白。结论 +Gz环境暴露可引起家兔颈部肌肉损伤,导致细胞凋亡,而固定家兔颈部受力环节可缓解肌肉损伤症状;+Gz环境暴露导致的细胞凋亡可能与线粒体功能相关。

+Gz暴露  /  飞行员  /  颈部伤病  /  肌肉损伤

Objective To study the mechanism of +Gz environment exposure induced neck muscle injury in rabbits. Methods A total of 20 male rabbits were randomized into the negative control group, +Gz exposure group, +Gz exposure plus neck loading group, and +Gz exposure plus neck loading plus fixation group (5 in each group). The animal centrifuge was used to simulate the +Gz exposure environment. Rabbits in the +Gz exposure group, +Gz exposure plus neck loading group, and +Gz exposure plus neck loading plus fixation group were exposed for 4 weeks in a +6 Gz environment. Before and after the experiment, the rabbits in each group were photographed by X-ray. At the time of 0-, 1-, 2-, 3- and 4-week exposure, ear vein blood was collected from rabbits in each group, and ELISA was performed to detected plasma leveles of lactate dehydrogenase (LDH), creatine kinase (CK) and reactive oxygen species (ROS). After exposure, the trapezius muscle of rabbits was taken for HE staining to detect the muscle morphology.The apoptosis level was detected by TUNEL in the paraffin section of the rabbit trapezius muscle. The expressions of apoptosis-related proteins Bax and Bcl-2 in the trapezius muscle of rabbits were detected by Western blotting. The rabbit's neck trapezius muscle was taken to conduct proteomics testing, select differentially expressed proteins, and perform protein function cluster analysis. Results There was no significant difference in the morphology of the cervical vertebra in each group before and after+Gz exposure. Compared with negative control group, plasma levels of LDH, CK, and ROS in +Gz exposure group, +Gz exposure plus neck loading group, and +Gz exposure plus neck loading plus fixation group significantly increased (P<0.05). Compared with negative control group, the ratio of apoptosis protein Bax/Bcl-2 in +Gz exposure group significantly increased (P<0.05), the number of positive apoptosis cells detected by TUNEL significantly increased (P<0.05). Compared with +Gz exposure group, the+Gz exposure plus neck loading group showed an increased Bax/Bcl-2 ratio (P<0.05) and more TUNEL positive cells (P<0.05).Compared with +Gz exposure plus neck loading group, the ratio of apoptosis protein Bax/Bcl-2 and the positive number of apoptosis were decreased in +Gz exposure plus neck loading plus fixation group, but there were no statistically significant differences (P>0.05).A total of 600 proteins were significantly altered in +Gz exposure group compared to negative control group, and GO functional clustering analysis of the differential proteins revealed significant enrichment of proteins associated with mitochondrial function. Conclusion Exposure to the +Gz environment can cause neck muscle injury and cell apoptosis in rabbits. The muscle injury symptoms can be improved by fixing the stress link of the rabbit neck. Cell apoptosis induced by +Gz exposure may be related to mitochondrial function.

+Gz exposure  /  pilot  /  neck injury  /  muscle injury
李卓, 李曦, 潘益凯, 李程飞, 范洁怡, 铁娅滕, 赵星成, 王永春. +Gz暴露诱发家兔颈部骨骼肌损伤的机制研究. 解放军医学杂志, 2022 , 47 (2) : 136 -142 . DOI: 10.11855/j.issn.0577-7402.2022.02.0136
Zhuo Li, Xi Li, Yi-Kai Pan, Cheng-Fei Li, Jie-Yi Fan, Ya-Teng Tie, Xing-Cheng Zhao, Yong-Chun Wang. Mechanism of cervical skeletal muscle injury induced by +Gz exposure in rabbits[J]. Medical Journal of Chinese People’s Liberation Army, 2022 , 47 (2) : 136 -142 . DOI: 10.11855/j.issn.0577-7402.2022.02.0136
颈椎病以颈椎退行性改变、颈部肌群慢性劳损为主,目前呈现发病率高、年轻化的临床特点[1-2]。颈部肌群的力量相对较弱,而颈椎为脊柱中不稳定的节段,因此颈部是人体解剖结构中的薄弱环节。不良的体态坐姿等生活习惯会导致颈部承受更多压力,诱发颈部病变,轻型病变涉及肌肉、韧带等,重型病变则涉及骨骼、神经、血管等[3]。颈椎病在战斗机飞行员人群中是一种常见疾病,与其特殊工作环境关系密切[4]。现代高性能战斗机在机动飞行中所产生的正加速度(+Gz)具有高G值、快增长率、持续时间长、反复出现等特点,使得高性能战斗机飞行员在高空飞行环境中颈部不断承受高载荷,而头盔及各种通信装备的附加重量也增加了颈部的负荷[5-6],因此,易导致飞行员发生急性颈部损伤。另外,若频繁经受持续性高+Gz作用,则会导致颈部肌群劳损加剧,致使颈椎间盘的退行性病变提前发生[7]。但是,目前有关持续性+Gz环境暴露对颈部肌肉损伤的研究较少,颈部组织病理的发生与+Gz暴露的关系有待深入探讨。本研究模拟持续性+Gz暴露环境,探讨+Gz暴露诱发家兔颈部肌肉损伤的机制,以期为飞行员颈部损伤的防护与治疗提供理论依据。
活性氧(ROS,#MM-82597002)、乳酸脱氢酶(LDH,#MM-8261302)、肌酸激酶(CK,MM-33553202)试剂盒购自上海酶联生物科技有限公司;Bcl-2抗体(#15071)、IgG抗体(#7076)、Mouse IgG抗体(#7076)、Rabbit IgG抗体(#7074)购自美国CST公司;Bax抗体(ab32503)购自英国Abcam公司;GAPDH抗体(#sc-73322)购自美国Santa Cruz公司。动物离心机由空军军医大学航空航天医学系提供;Tanon电泳仪及凝胶成像系统(#5200)购自上海天能科技有限公司;荧光显微镜(ECLIPSE C1)购自日本Nikon公司;光学显微镜(BX53)购自日本Olympus公司。
健康雄性家兔20只,购自空军军医大学动物中心,随机分为正常对照组(正常饲养环境)、+Gz暴露组(+6 Gz环境暴露)、+Gz暴露+颈部负重组(+6 Gz环境暴露,同时对颈部施加负重)、+Gz暴露+颈部负重+固定组(+6 Gz环境暴露,同时进行颈部负重和颈腰部固定),每组5只。家兔颈部负重方案参照飞行员头盔占头部重量百分比(约为30%),确定家兔头部负重约100 g。本研究经空军军医大学动物福利与伦理委员会批准(IACUC-20201006),实验过程符合国家和单位有关实验动物的管理和使用规定。
+Gz环境模拟:采用动物离心机模拟+Gz暴露,动物离心机半径为2 m,可模拟的+Gz范围为+1~+12 Gz,利用半圆柱动物舱承载动物,水平固定于离心机转臂,动物头部朝向离心机轴心。+Gz暴露加速度G值为+6 Gz,每天连续暴露3 min,持续暴露4周(图1)。
暴露前及暴露1、2、3、4周,使用肝素抗凝管采集家兔耳缘静脉血,1500 r/min离心,收集血浆。暴露结束24 h后,麻醉处死家兔,备皮后手术台固定,取左右侧颈部斜方肌,生理盐水冲洗,于4%多聚甲醛溶液和液氮中保存。
暴露前和暴露4周后,耳缘静脉注射麻醉家兔,于相同条件、相同体位下拍摄颈部侧位X线片,观察颈部病变情况。
采用ELISA试剂盒检测家兔血浆LDH、CK、ROS水平,严格按照试剂盒说明书步骤操作。
家兔肌肉组织在4%多聚甲醛溶液中固定48 h后,流水冲洗6 h,常规石蜡包埋、切片,严格按照HE染色试剂盒说明书步骤操作,光学显微镜下观察骨骼肌纤维走向、排布等形态变化。
严格按照TUNEL试剂盒说明书操作。石蜡切片进行脱蜡脱水,滴加蛋白酶K,37 ℃孵育30 min;PBS清洗5 min×5次,37 ℃下避光孵育TUNEL检测液1 h;PBS清洗5 min×5次,封片剂封片。荧光显微镜下观察细胞凋亡情况。
取50 mg液氮保存的家兔斜方肌组织,加入300 μl裂解液混合物,冰浴匀浆,离心取上清,行BCA蛋白定量。采用SurePAGETM(Genscrip,#M00657)凝胶电泳分离蛋白,湿转法转膜,室温下5% BSA封闭1 h,加入Bcl-2(1∶1500)、Bax(1∶1000)、GAPDH(1∶5000)一抗4 ℃孵育过夜;次日TBST清洗,加入Mouse IgG抗体(1∶8000)或Rabbit IgG抗体(1∶8000)室温孵育1.5 h;TBST清洗,加入ECL发光液,凝胶成像系统显影成像,使用ImageJ软件分析结果。
家兔斜方肌肌肉组织于液氮中保存,送至杭州景杰生物公司行多肽体外标记定量技术(tandem mass tags,TMT)蛋白组学检测。使用GO数据库(gene ontology,GO)将鉴定到的差异蛋白进行基因本体论富集分析,用于表述蛋白的各种功能属性。
采用GraphPad Prism 9.0软件进行统计分析并制图。计量资料以$\bar{x}±s$表示,多组间比较采用单因素方差分析(one-way ANOVA,Turkey法),进一步两两比较采用LSD-t检验。P<0.05为差异有统计学意义。
X线观察家兔侧位状态下颈椎形态变化,结果显示,与暴露前相比,各组家兔在+Gz暴露4周后,颈椎骨的生理弯曲、骨间隙等无显著病理性改变;但与暴露前相比,暴露4周后+Gz暴露组、+Gz暴露+颈部负重组和+Gz暴露+颈部负重+固定组家兔软组织影变深,提示肌肉等软组织可能出现增厚增生变化(图2)。
ELISA检测结果显示,家兔血浆LDH、CK及ROS水平与+Gz暴露时间呈正相关,+Gz暴露4周后,与正常对照组[LDH:(27.7±3) ng/ml;CK:(78.4±2.9) ng/ml;ROS:(120.8±13.6) U/ml]相比;+Gz暴露组[LDH:(37.7±1.7) ng/ml;CK:(120.2±5.3) ng/ml;ROS:(186.7±17) U/ml]、+Gz暴露+颈部负重组[LDH:(46.9±1.5) ng/ml;CK:(136.7±1.2) ng/ml;ROS:(219.1±11.5) U/ml]、+Gz暴露+颈部负重+固定组[LDH:(41.9±2.5) ng/ml;CK:(127±5) ng/ml;ROS:(208.7±5.9) U/ml]血浆LDH、CK和ROS水平升高,差异有统计学意义(P<0.05),且+Gz暴露+颈部负重组血浆LDH、CK及ROS水平最高(图3)。
HE染色结果显示,正常对照组骨骼肌纤维排列紧致有序无皱褶,且走向清晰;与正常对照组比较,+Gz暴露组、+Gz暴露+颈部负重组和+Gz暴露+颈部负重+固定组骨骼肌纤维均发生扭曲紊乱;与+Gz暴露+颈部负重组比较,+Gz暴露+颈部负重+固定组骨骼肌纤维相对清晰完整,肌纤维扭曲紊乱情况有所减轻(图4)。
TUNEL染色结果显示,+Gz暴露后,家兔斜方肌出现凋亡细胞;与+Gz暴露组相比,+Gz暴露+颈部负重组斜方肌TUNEL阳性细胞数明显增多(P<0.05);与+Gz暴露+颈部负重组比较,+Gz暴露+颈部负重+固定组斜方肌TUNEL阳性细胞数有所减少,但差异无统计学意义(P>0.05,图5A)。
Western blotting检测凋亡相关蛋白Bax、Bcl-2的表达,结果显示,与正常对照组比较,+Gz暴露组Bax/Bcl-2比值明显增高(P<0.05);与+Gz暴露+颈部负重组比较,+Gz暴露+颈部负重+固定组Bax/Bcl-2比值降低,但差异无统计学意义(P>0.05,图5B)。
为了发现在+Gz环境下引起家兔颈部肌肉损伤的潜在靶点,对正常对照组与+Gz暴露组进行蛋白组学筛查,以蛋白差异表达量变化超过1.3倍作为显著上调的变化阈值,小于1.3倍作为显著下调的变化阈值,结果显示,与正常对照组比较,+Gz暴露组中共600个蛋白发生显著变化,其中上调475个,下调125个(图6A);对差异蛋白进行GO功能聚类分析,结果显示,差异蛋白显著富集于与线粒体功能相关的蛋白,包括线粒体膜蛋白、线粒体呼吸链相关蛋白等(图6B)。
高性能战斗机飞行员颈部经常暴露在高惯性力作用下,具有武器瞄准、通讯、夜视等多功能的现代头盔装置改变了驾驶舱的人体工程学。头盔系统重量的增加及其重心的移动增加了驾驶操作过程中移动头部时对力量的需求,加重了+Gz负荷对颈椎和肌肉的有害影响,这使得战斗机飞行员的颈部面临更高的受伤风险,不仅慢性疲劳、慢性劳损风险增加,而且在极端飞行过程中,对颈部的急性损伤也不容忽视[8-10]。重力会影响物理和生物过程,在人体组织发育和动态平衡过程中起着关键作用,地球上的生命在进化过程中已经适应了正常的重力环境,然而由于人体工作环境(高速环境、宇航环境等)的变化,微重力和超重力会对机体造成不同的影响,这已引起人们的极大关注。有研究发现,由于暴露在微重力或超重力的特定环境条件下,宇航员和飞行员在飞行后椎间盘退行性改变的发生率增高[11-12]。但在飞行过程中使颈部受力环节尽可能保持稳定可显著降低损伤率,减缓损伤发生进程。本研究观察了+Gz暴露对家兔颈部肌肉的影响,并对家兔头部施加负重及采用物理手段稳定颈部,X线片显示,在+6 Gz暴露4周后,各组家兔的颈椎形态没有显著差异,在此实验条件下连续+Gz暴露未对骨骼形态造成影响。但对颈部肌肉的组织病理学、分子生物学检测发现,+Gz暴露+颈部负重组较正常对照组颈部肌肉变化明显,包括血浆CK、LDH、ROS水平增高,肌纤维紊乱,以及肌细胞凋亡增多等,而这些指标的变化反映了肌肉代谢、超微结构的改变,提示肌肉已经开始出现损伤,因此+Gz环境所引发的肌肉损伤、肌细胞凋亡应受到关注。
有研究发现,太空飞行会对机体生理产生复杂的影响,可引起体内代谢、免疫等多方面的调节反馈,其中线粒体能量代谢、肝脏脂质代谢等变化显著,许多蛋白、microRNA等代谢物会影响线粒体的能量代谢,引发连锁反应[13-14]。压力在这些变化中起着至关重要的作用。虽然太空宇航飞行与军事航空飞行环境不尽相同,但可以确定的是压力、重力等特殊力学环境会对人体造成影响,引发一系列生理病理反应,细胞凋亡即为其中之一。体内一系列生化反应都可能引发细胞凋亡,如炎症反应、氧化应激、DNA损伤、线粒体功能紊乱等[15-17]。Bcl-2蛋白家族是与细胞凋亡高度相关的一类蛋白,该家族的一些成员(Bcl-2、Bcl-xL、Bax等)主要位于线粒体外膜上,可能通过与线粒体膜通透性转换孔(mitochondrial permeability transition pore,mPTP)成分相互作用而调节mPTP的开放和关闭,或自身聚集形成通道,从而影响细胞凋亡[18-20],而线粒体膜通透性转换孔道复合物是线粒体接受各种信号的感受器,被认为是细胞生死的开关。ROS也是mPTP开放的强诱导剂,研究证实,在运动疲劳、衰老和缺血/再灌注等病理情况下均伴有呼吸链电子漏偏高和线粒体自由基代谢失调现象,过度的ROS持续积累可导致mPTP持续开放[21-23]。本研究发现,在+Gz暴露后,家兔血液中ROS含量明显增加,TUNEL实验证实了在此环境下细胞发生凋亡,通过Western blotting检测家兔颈部肌肉中Bcl-2和Bax蛋白的表达,进一步验证了细胞凋亡的发生。通过对正常环境和+Gz环境下两组样本进行蛋白组学检测,发现两者之间存在较大差异,+Gz环境引起了家兔颈部肌肉蛋白表达的改变;对差异表达蛋白进行GO功能聚类分析,结果显示,差异表达蛋白显著富集于与线粒体功能相关的蛋白,结合前文所述,提示在+Gz环境下发生的细胞凋亡可能与线粒体功能变化高度相关,但+Gz环境所引起的蛋白变化及影响线粒体功能改变的具体机制有待进一步深入研究。
战斗机飞行员人群中颈椎病是高发疾病,对飞行员的工作能力和工作状态造成了一定影响,严重情况下甚至可缩短飞行员的职业生涯,造成非战斗减员等损失。目前针对飞行员颈椎病的预防手段主要从两方面考虑:一为尽量设计符合人体工程学的装备,使颈部受力更加符合人体解剖学、运动生物力学规律,本研究结果也体现了稳定颈部关键受力环节带来的益处;二为飞行员日常训练中专项强化颈腰肌群,提高对抗负荷的工作能力。目前临床对颈椎病有手术、止痛、理疗、康复训练等多种对症治疗方法,但治疗效果因人、因病情而异[24-26]。本研究通过动物实验模拟飞行员高Gz的飞行环境,希望通过了解这一特殊环境下的肌肉损伤机制,为其治疗提供新视角。本研究结果显示,通过动物离心机模拟的+Gz环境可引起家兔颈部肌肉形态的变化,且颈部负重会恶化肌肉病变进程,而固定其主要受力环节可改善病理进程。本研究虽初步建立了+Gz环境下的动物肌肉损伤模型,通过分子生物学、组织病理学等手段观察了其表型变化,通过蛋白组学检测发现了+Gz环境这一特殊环境引起的肌肉蛋白改变,但对其关键作用靶点的筛选及深入验证尚有待进一步探讨。
综上所述,+Gz暴露可引起家兔颈部肌肉损伤,导致细胞凋亡,而通过物理手段固定家兔颈部受力环节可减轻肌肉损伤;+Gz环境暴露导致的细胞凋亡可能与线粒体功能相关,但其诱发细胞凋亡的具体机制有待进一步深入研究。
  • 空军装备课题(KJ20191A050359)
  • 空军军医大学效能提升计划(2019ZTA05-02)
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2022年第47卷第2期
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doi: 10.11855/j.issn.0577-7402.2022.02.0136
  • 接收时间:2021-07-08
  • 首发时间:2025-12-17
  • 出版时间:2022-02-28
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  • 收稿日期:2021-07-08
  • 录用日期:2021-09-06
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Air Force Equipment Project(KJ20191A050359)
空军装备课题(KJ20191A050359)
Air Force Military Medical University Effectiveness Improvement Plan(2019ZTA05-02)
空军军医大学效能提升计划(2019ZTA05-02)
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    空军军医大学航空航天医学系航空航天医学训练教研室,西安 710032

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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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