电针对前交叉韧带断裂大鼠炎症反应及cGAS/STING信号通路的影响

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中文摘要：目的:探究电针对前交叉韧带断裂后创伤性骨关节炎早期炎症反应与环状GMP-AMP合成酶(cGAS)/干扰素基因刺激因子(STING)信号通路的影响。方法:24只SD大鼠随机分为假手术组、前交叉韧带损伤(ACLT)组和电针组，每组8只。其中ACLT组和电针组实施前交叉韧带切断术。电针组大鼠予双侧"阳陵泉""血海""足三里""太溪"电针刺激，1次/d，每周干预5 d，连续2周，其余两组不做处理。2周后ELISA法检测大鼠血清白介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)水平;取大鼠左膝关节软骨组织切片分别进行番红O-固绿染色和苏木精-伊红(HE)染色，镜下观察软骨损伤程度并评分(Mankin's评分);RT-qPCR检测大鼠右膝关节软骨基质金属蛋白酶-13(MMP-13)mRNA、cGAS mRNA、STING mRNA表达水平。Western blotting检测大鼠右膝关节软骨MMP-13、cGAS、STING蛋白表达水平。结果:ACLT组大鼠血清IL-1β、TNF-α水平均高于假手术组(P＜0。05);电针组大鼠血清IL-1β、TNF-α水平均低于ACLT组(P＜0。05)。番红O-固绿染色显示假手术组大鼠软骨表面光滑;ACLT组大鼠软骨表面出现轻微缺损和细小裂层;电针组大鼠软骨表面较平滑，无明显裂层。HE染色显示假手术组软骨表面平整且光滑，软骨细胞均排列规整;ACLT组大鼠软骨层表面变薄;电针组大鼠软骨表面略粗糙。ACLT组大鼠Mankin's评分高于假手术组(P＜0。05);电针组大鼠Mankin's评分低于ACLT组(P＜0。05)。ACLT组大鼠MMP-13 mRNA、cGAS mRNA、STING mRNA相对表达量均高于假手术组(P＜0。05);电针组大鼠MMP-13 mRNA、cGAS mRNA、STING mRNA相对表达量均低于ACLT组(P＜0。05)。ACLT组大鼠MMP-13、cGAS、STING蛋白相对表达量均高于假手术组(P＜0。01);电针组大鼠MMP-13、cGAS、STING蛋白相对表达量均低于ACLT组(P＜0。01)。结论:电针可能通过下调cGAS/STING信号通路而减轻ACLT大鼠炎症反应，延缓关节软骨退行性改变。

外文标题：Effects of Electroacupuncture on Inflammatory Response and cGAS/STING Signaling Pathway in Rats with Anterior Cruciate Ligament Rupture

外文摘要：Objective:To investigate the progression of traumatic osteoarthritis after anterior cruciate ligament rupture(ACLT)and the effect of electroacupuncture on its early inflammatory response and cyclic GMP-AMP synthase(cGAS)/interferon gene stimulating factor(STING)signaling pathway.Methods:A total of 24 SD rats were randomly divided into sham operation group,anterior cruciate ligament injury(ACLT)group and electroacupuncture group,with 8 rats in each group.Among them,ACLT group and electroacupuncture group were subjected to ACL amputation.In the electroacupuncture group,rats were given bilateral electroacupuncture stimulation of"Yanglingquan"(GB 34),"Xuehai"(SP 10),"Zusanli"(ST 36)and"Taixi"(KI 3)once a day,and the intervention was carried out for 5 days per week.After 2 weeks,the rats'serum interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)levels were detected by ELISA;The cartilage tissue sections of the left knee joint were stained with sapphire red O-green and hematoxylin-eosin(HE),and the extent of the cartilage damage was observed and scored under the microscope(Mankin's score);RT-qPCR was performed to detect the expression levels of matrix metalloproteinase-13(MMP-13)mRNA,cGAS mRNA and STING mRNA in the cartilage of the right knee joint of rats.Western blotting was performed to detect the expression levels of MMP-13,cGAS and STING proteins in the cartilage of the right knee joint of rats.Results:The ACLT group showed higher serum IL-1β and TNF-α levels than sham operation group(P＜0.05).The electroacupuncture group showed lower serum IL-1β and TNF-α levels than ACLT group(P＜0.05).The surface of cartilage of rats in the sham operation group was smooth;The surface of cartilage of rats in the ACLT group showed slight defects and small cracks;The surface of cartilage of rats in the electro-acupuncture group was smooth without obvious cracks.HE staining showed that the surface of cartilage of rats in the sham operation group was smooth and even,and the chondrocytes were arranged in a regular way;The surface of cartilage of rats in the ACLT group was thinned;The surface of cartilage of rats in the electroacupuncture group was a little bit rough;The ACLT group showed higher Mankin's score than sham operation group(P＜0.05);The electroacupuncture group showed lower Mankin's score than ACLT group(P＜0.05).The ACLT group showed higher relative expression of MMP-13 mRNA,cGAS mRNA and STING mRNA than sham operation group(P＜0.05);The electroacupuncture group showed lower relative expression of MMP-13 mRNA,cGAS mRNA and STING mRNA than ACLT group(P＜0.05).The ACLT group showed higher relative expression of MMP-13,cGAS and STING proteins than sham-operated group(P＜0.01);The electroacupuncture group showed lower relative expression of MMP-13,cGAS and STING proteins than ACLT group(P＜0.01).Conclusion:Electroacupuncture may reduce inflammatory response and delay articular cartilage degeneration in ACLT rats by down-regulating cGAS/STING signaling pathway.

外文关键词：

anterior cruciate ligament injuryanterior cruciate ligament dissectionelectroacupunctureinflammatory responsearticular cartilage,cGAS/STING signaling pathwayrats

作者：

贾飞阳、李萌萌、王金玲、刘静、黄夏荣、孙光华、钟培瑞、彭婷、曾国锋、周君

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作者单位：

南华大学衡阳医学院附属第一医院,湖南衡阳 421001

四川大学华西医院康复医学四川省重点实验室,四川成都 610041

关键词：

前交叉韧带损伤创伤性骨关节炎电针炎症反应关节软骨 cGAS/STING信号通路大鼠

基金：

国家自然科学基金南华大学附属第一医院4310计划南华大学校级重点项目湖南省自然科学基金湖南省临床医疗技术创新引导项目

项目编号：

8197391720214310NHYCG07USCKF201902K022023JJ305432021SK51805

出版年：

2024

DOI：

10.13862/j.cn43-1446/r.2024.05.001

中医药导报

湖南省中医药学会湖南省中医管理局

中医药导报

CSTPCD

影响因子：0.952

ISSN：1672-951X

年,卷(期)：2024.30(5)

参考文献量7