Exploration of gastrocnemius injury and its mechanism in rats with syndrome of spleen deficiency

FAN Fangxin; ZHU Haoru; QIAN Meng; SHI Tuo; ZHU Sitai; XIE Ming

doi:10.3969/j.issn.1006-2157.2023.01.013

您当前的位置：

首页 >

文章列表页 >

Exploration of gastrocnemius injury and its mechanism in rats with syndrome of spleen deficiency

Experimental Studies | 更新时间：2023-02-03

- Exploration of gastrocnemius injury and its mechanism in rats with syndrome of spleen deficiency
- Journal of Beijing University of Traditional Chinese Medicine Vol. 46, Issue 1, Pages: 77-86(2023)
- 作者机构：
  
  北京中医药大学中医学院　北京　100029
- 作者简介：
  
  Prof. XIE Ming, Ph. D. , Doctoral Supervisor. School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, No.11, Beisanhuan Donglu Road, Chaoyang District, Beijing, 100029. E-mail: xieming603@263.net
- 基金信息：
  
  National Natural Science Foundation of China(81173193)
- DOI：10.3969/j.issn.1006-2157.2023.01.013
  CLC： R285.5
- Received：28 February 2022，
  
  Online First：02 November 2022，
  
  Published：30 January 2023
- 稿件说明：
移动端阅览
FAN Fangxin, ZHU Haoru, QIAN Meng, et al. Exploration of gastrocnemius injury and its mechanism in rats with syndrome of spleen deficiency[J]. Journal of Beijing University of Traditional Chinese Medicine, 2023, 46(1): 77-86.
DOI：

FAN Fangxin, ZHU Haoru, QIAN Meng, et al. Exploration of gastrocnemius injury and its mechanism in rats with syndrome of spleen deficiency[J]. Journal of Beijing University of Traditional Chinese Medicine, 2023, 46(1): 77-86. DOI： 10.3969/j.issn.1006-2157.2023.01.013.

摘要

目的

观察脾虚证模型大鼠腓肠肌损伤情况并对其机制进行研究。

方法

将雄性Wistar大鼠(SPF/VAF级)按体质量随机分为对照组和模型组，模型组采用饮食失节联合游泳劳倦法复制脾虚证模型，对照组予以正常饲养。定期观察并记录2组大鼠的外观表征、体质量、饮食量和粪便含水率。实验第2、3、4周末，测定各组大鼠的游泳力竭时间、抓握力、尿D－木糖排泄率、脾脏指数、胸腺指数及腓肠肌湿质量，HE染色观察腓肠肌病理结构，酶联免疫吸附测定法检测血清中胃动素、胃泌素、白细胞介素－1β(IL-1β)、白细胞介素－6(IL-6)、肿瘤坏死因子-α(TNF-α)和腓肠肌组织中柠檬酸合酶、Ca

2＋

－ATP酶、IL-1β、IL-6、TNF-α含量。实验第4周末，用免疫组织化学法检测腓肠肌IL-1β、IL-6、TNF-α阳性表达水平，免疫荧光法检测腓肠肌切割型天冬氨酸蛋白水解酶3(cleaved-caspase 3)阳性细胞。

结果

与对照组比较，第2～4周模型组大鼠体质量、饮食量、游泳力竭时间、抓握力、腓肠肌湿质量和尿D－木糖排泄率下降(

0.05，

0.01)，粪便含水率增加(

0.01)；第3～4周模型组大鼠血清胃泌素下降(

0.05，

0.01)；第2周模型组大鼠腓肠肌湿质量/体质量和血清IL-6增加(

0.05，

0.01)；第4周模型组大鼠腓肠肌湿质量/体质量、脾脏指数、胸腺指数、腓肠肌柠檬酸合酶和Ca

2＋

－ATP酶水平、血清胃泌素、IL-1β和TNF-α含量下降(

0.01)，腓肠肌cleaved-caspase 3、IL-1β、IL-6、TNF-α含量增加(

0.01)。

结论

饮食失节联合游泳劳倦法复制的脾虚证模型大鼠具有中医脾虚证的外观表征和实验室相关指标的变化特点，腓肠肌出现肌力减退和肌萎缩，并随造模时间延长而加重，伴有全身免疫功能减退与血清、腓肠肌炎性因子含量增加，提示脾虚证骨骼肌受损涉及线粒体损伤和细胞凋亡，并与炎性机制有关。

Abstract

Objective

To investigate skeletal muscle injury and its mechanism in a rat model of spleen deficiency syndrome.

Methods

Male Wistar rats(SPF/VAF) were randomly divided into control group and model group according to body weight. The "improper diet combined with exhaustive swimming" method was used to duplicate the rat model with spleen deficiency syndrome. The control group was fed normally. We regularly recorded the appearance

body weight

dietary intake

and excrement water content of the two groups of rats. At the end of Week 2

and 4

the swimming exhaustion time

grip strength

urine D-xylose excretion rate

spleen index

thymus index

and gastrocnemius wet weight were measured

and the gastrocnemius was subjected to pathological examination by HE staining. Besides

at the end of Week 2

and 4

we used enzyme-linked immunoassay to determine the levels of motilin

gastrin

interleukin-1β (IL-1β)

interleukin-6 (IL-6)

and tumor necrosis factor-α (TNF-α) in serum

and the levels of citrate synthase

-ATPase

IL-1β

IL-6

and TNF-α in the gastrocnemius. At the end of Week 4

immunohistochemistry method was used to measure the positive expression of IL-1β

IL-6

and TNF-α

and immunofluorescence was used to detect the cells that were positive for cysteine aspartate proteolytic enzyme 3 (cleaved-caspase 3) in the gastrocnemius.

Results

Compared to the control group

the body weight

dietary intake

swimming exhaustion time

grip strength

gastrocnemius wet weight

and urine D-xylose excretion rate in the model group were significantly decreased from Week 2 to 4 (

0.05

0.01)

while excrement water content increased (

0.01). Serum gastrin in the model group was also significantly decreased at Week 3 and 4 (

0.05

0.01). Besides

the gastrocnemius wet weight to body weight ratio and serum IL-6 of the rats in the model group were increased at Week 2 (

0.05

0.01). At the end of Week 4

the gastrocnemius wet weight to body weight ratio

spleen index

thymus index

citrate synthase and Ca

-ATPase in the gastrocnemius

the levels of gastrin

IL-1β

and TNF-α in the serum of the model group decreased significantly (

0.01); and the levels of cleaved-caspase 3

IL-1β

IL-6

and TNF-α in the gastrocnemius increased significantly (

0.01).

Conclusion

The spleen deficiency syndrome rat model induced by "improper diet combined with exhaustive swimming" method in accordance with the appearance of traditional Chinese medicine syndrome of spleen deficiency and related laboratory indexes. The muscle strength and atrophy of the gastrocnemius was decreased with the modeling time. The systemic immune function of the model rats decreased

while the protein expression of systemic and gastrocnemius inflammatory factors increased. We presume that the skeletal muscle damage involves mitochondrial injury and apoptosis in the spleen deficiency syndrome rat model but is related to inflammatory mechanism.

关键词

Keywords

references

郭丽娜 . 脾虚证能量代谢障碍与线粒体COXⅠ、COXⅣ表达的相关性研究 [D]. 广州：广州中医药大学， 2015 .

衣雪浩 . 健脾补糖对过度训练大鼠骨骼肌GLUT4和相关信号转导的影响 [D]. 沈阳：辽宁中医药大学， 2006 .

宋雅芳，胡任飞，刘友章，等 . 健脾祛湿方对重症肌无力模型大鼠骨骼肌线粒体及神经肌肉接头处超微结构的影响 [J]. 中药药理与临床， 2010 ， 26 ( 1 )： 65 － 68 .

林白阳 . 老年COPD患者血清IL-6、IL－10水平与肌少症的关系 [D]. 太原：山西医科大学， 2019 .

熊杉 . 针推膀胱经治疗慢性疲劳综合征及对IL-6、IFN－γ、TNF-α等细胞因子的影响 [D]. 武汉：湖北中医药大学， 2013 .

张广霞 . 健脾－健脾渗湿－健脾升阳方与脾虚证的相关性研究 [D]. 北京：北京中医药大学， 2016 .

王洪海 . 四君子汤　柴疏四君汤　柴胡疏肝散作用于脾虚证大鼠模型的生物效应比较 [D]. 北京：北京中医药大学， 2007 .

JULIO-PIEPER M , HYLAND NP , BRAVO JA , et al . A novel role for the metabotropic glutamate receptor－7: mo-dulation of faecal water content and colonic electrolyte transport in the mouse [J]. Br J Pharmacol ， 2010 ， 160 ( 2 )： 367 － 375 .

LAMOU B , TAIWE GS , HAMADOU A , et al . Anti-oxidant and antifatigue properties of the aqueous extract of moringa oleifera in rats subjected to forced swimming endurance test [J/OL]. Oxid Med Cell Longev , 2016 ， 2016 ： 3517824 [ 2021－03－24 ]. https：//www.hindawi.com/journals/omcl/2016/3517824/ https：//www.hindawi.com/journals/omcl/2016/3517824/ .

武曲星，王攀，王冬芝，等 . 免疫性血小板减少症气不摄血证小鼠模型血管内皮活性物质变化动态研究 [J]. 中国中医药信息杂志， 2019 ， 26 ( 12 )： 56 － 61 .

李亚欢，张冬梅，王淑艳，等 . 脾虚证动物模型研制概况 [J]. 中医药导报， 2019 ， 25 ( 1 )： 100 － 102 .

唐晓桐，曲笑锋，于艳辉，等 . 养脾丸对脾虚证大鼠模型胃肠及免疫功能的影响 [J]. 人参研究， 2019 ， 31 ( 6 )： 20 － 23 .

屈小虎，黄玲，陈慧，等 . 脾气虚证和脾不统血证模型大鼠脾主肌肉实质的探讨 [J]. 中国中西医结合杂志， 2019 ， 39 ( 2 )： 217 － 222 .

李松霖 . 半夏泻心汤联合西药治疗消化性溃疡的疗效及对血浆胃动素、胃泌素的影响 [J]. 中国处方药， 2019 ， 17 ( 11 )： 145 － 146 .

高小玲，郭文峰，李茹柳，等 . 四君子汤对脾虚大鼠尿木糖排泄率及肠黏膜ATP的影响 [J]. 中药材， 2009 ， 32 ( 8 )： 1242 － 1245 .

刘海涛，施家希，黄张杰，等 . 补中益气汤不同配伍对脾虚大鼠免疫器官及小肠黏膜转运体蛋白表达的影响 [J]. 中药材， 2017 ， 40 ( 2 )： 466 － 469 .

陈旻丹，胡京红，马捷，等 . 脾虚四号方含药血清对功能性腹泻脾虚证大鼠离体平滑肌细胞的影响 [J]. 北京中医药大学学报， 2017 ， 40 ( 4 )： 297 － 301 .

谢凌坚，张珊，刘素娟，等 . AMPKα2在高强度间歇运动激活小鼠骨骼肌细胞自噬过程中的作用 [J]. 中国运动医学杂志， 2018 ， 37 ( 4 )： 309 － 314 .

BLOXHAM DP , PARMELEE DC , KUMAR S , et al . Primary structure of Porcine heart citrate synthase [J]. Proc Natl Acad Sci USA ， 1981 ， 78 ( 9 ): 5381 － 5385 .

段永强，梁玉杰，成映霞，等 . 脾气虚大鼠骨骼肌组织Ca 2＋ /CaM信号通路关键分子动态表达规律 [J]. 中国老年学杂志， 2015 ， 35 ( 15 )： 4127 － 4130 .

卢智勇 . Caspase－3细胞定位信号的检测与分析 [D]. 重庆：重庆医科大学， 2008 .

费文婷，侯燕，王玉杰，等 . 玛咖性温健脾及对脾虚小鼠线粒体能量代谢酶的影响 [J]. 北京中医药大学学报， 2018 ， 41 ( 7 )： 559 － 566 .

管春彦，牛慧彦 . 肿瘤恶液质诱导骨骼肌氧化代谢调节作用的研究进展 [J]. 肿瘤代谢与营养电子杂志， 2018 ， 5 ( 4 )： 342 － 347 .

田振军，贺志雄，蔡梦昕 . 运动干预骨骼肌内分泌功能 [J]. 生理科学进展， 2013 ， 44 ( 4 )： 275 － 280 .

ZHAO HJ , WANG Y , SHAO YZ , et al . Oxidative stress-induced skeletal muscle injury involves in NF－κB/P53－activated immunosuppression and apoptosis response in copper (Ⅱ) or/and arsenite-exposed chicken [J]. Chemosphere ， 2018 ， 210 ： 76 － 84 .

钱桂凤 . 针灸治疗慢性疲劳综合征的疗效观察及对大鼠抗氧化、免疫机制研究 [D]. 广州：广州中医药大学， 2018 .

范圣东 . 人参皂苷Rb1改善术后疲劳模型大鼠骨骼肌炎症反应和肌肉萎缩的机制研究 [D]. 合肥：安徽医科大学， 2019 .

王宁 . 大蒜素对兔急性下肢缺血－再灌注损伤后组织中IL－1、IL-6、IL－8含量的影响 [D]. 沈阳：辽宁中医药大学， 2011 .

赵杨，孙波，杜书章 . 芦荟素抑制P65的磷酸化对大鼠脑缺血再灌注诱导的组织损伤和炎症的调节作用 [J]. 中国免疫学杂志， 2019 ， 35 ( 17 )： 2084 － 2088 .

THOMA A , LIGHTFOOT AP . NF－κB and Inflammatory Cytokine Signalling: Role in Skeletal Muscle Atrophy [J]. Adv Exp Med Biol ， 2018 ， 1088 ： 267 － 279 .

祝琳 . 温针灸联合止痛通痹液治疗腰椎间盘突出症的临床观察及对血清TNF-α、IL-1β、IL-6 PGE 2 水平的影响 [J]. 中国中医药科技， 2019 ， 26 ( 2 )： 162 － 165 .

马晓玮，张天泰，杜冠华 . 新型非甾体抗炎药水杨酸甲酯糖苷抗神经小胶质细胞炎症作用的研究 [J]. 中国新药杂志， 2014 ， 23 ( 6 )： 654 － 659 .

白爱平 . 肠黏膜非免疫细胞在炎症性肠病发病中的作用 [J]. 世界华人消化杂志， 2010 ， 18 ( 19 )： 2020 － 2023 .

周迪，陈章健，胡贵平，等 . 纳米二氧化钛亚急性经口暴露对大鼠氧化/抗氧化生物标志和炎性因子的影响 [J]. 北京大学学报(医学版) ， 2020 ， 52 ( 5 )： 821 － 827 .

Views

331

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Effects and mechanism of the Tiansui Tongluo Formula in treating cerebral small vessel disease model rats by regulating miR-143-3p/NRG-1 axis

Liqi Huoxue Dripping Pill improves myocardial ischemia-reperfusion injury in rats by inhibiting apoptosis by regulating the HIF-1α/BNIP3 signaling pathway

Exploring the mechanism of regulating Bcl-2/Bax pathway by the therapy of promoting blood circulation and nourishing yin to improve spermatogenesis in rats with varicocele

Effect of therapeutic principle of "cultivating soil to restrict water" on ATP content and mitochondrial apoptosis pathway in rats with uremic cardiomyopathy

Protective effect of Honghua（Safflower,Flos Carthami） on acute liver injury induced by carbon tetrachloride in rats and mechanism study

Related Author

LIU Yuxuan

GUAN Dongsheng

ZHAO Huanghong

LIU Tianjing

CHEN Xiangyun

TANG Xiaofang

LU Yuxin

LI Yaofeng

Related Institution

Second Clinical Medical College，Henan University of Chinese Medicine

Department of Neurology，Henan Provincial Hospital of Traditional Chinese Medicine

School of Basic Medicine, Guizhou University of Traditional Chinese Medicine

The Second Clinical Medical College, Guangzhou University of Chinese Medicine

Zhuhai Hospital, Guangdong Provincial Hospital of Chinese Medicine

AI问答

Postal code：100029
Tel：(010)53911857 Email：jbutcm@bucm.edu.cn
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备09064830号-19 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰