骶神经电刺激对急性脊髓损伤大鼠肠黏膜免疫屏障功能的影响*

doi:10.12047/j.cjap.6023.2020.114

中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (6): 539-543.doi: 10.12047/j.cjap.6023.2020.114

骶神经电刺激对急性脊髓损伤大鼠肠黏膜免疫屏障功能的影响^*

白春宏^1△, 张文丽², 户士忠³

1.武警特色医学中心骨科, 天津 300162;
2.华北理工大学综合测试分析中心, 唐山 063000;
3.武警天津总队第一支队卫生处, 天津 300222

收稿日期:2020-03-06 修回日期:2020-11-25 出版日期:2020-11-28 发布日期:2021-03-15
通讯作者: ^△Tel: 15822960439; E-mail: 846556756@ qq.com
基金资助:
^*国家自然科学青年基金资助项目(81101441);天津市企业博士后创新项目择优资助(一等);武警后勤学院基础研究项目(WHJ2016017)

Sacral nerve stimulating on intestinal mucosal immune barrier function of rats with acute complete spinal cord injury

BAI Chun-hong^1△, ZHANG Wen-li², HU Shi-zhong³

1. Orthopaedics Department, Characteristic medical center of the Chinese people's Armed police Force, Tianjin 300162;
2. Comprehensive Testing and Analytical Center of North China University of Science and Technology, Tangshan 063000;
3. Health Department of the First Detachment of the Armed Police Tianjin Corps, Tianjin 300222, China

Received:2020-03-06 Revised:2020-11-25 Online:2020-11-28 Published:2021-03-15

摘要/Abstract

摘要： 目的:观察骶神经根电刺激对急性完全性脊髓损伤大鼠肠黏膜免疫屏障功能的影响。方法:56只Wistar大鼠分为正常组(SG=8只)、脊髓损伤组(CG=24只)和骶神经电刺激组(EG=24只)。CG、EG (分别设24、48和72 h 组,8只/组)。CG、EG大鼠均于全麻下用Fehlings法98 g动脉瘤夹横行钳夹行脊髓损伤并植入骶神经刺激电于右侧骶3神经孔。CG不进行干预,EG行骶神经电刺激。刺激参数:电压4 V,波宽210 μs,频率15 Hz,刺激10 s,间歇20 s。每次持续10 min,间歇10 min,共2 h。早8:00-10:00,晚6:00-8:00两次。分组采集标本,进行HE染色光镜、透射电镜组织学观察、Western blot 方法检测组织锌指蛋白A20、NOD2和CD68蛋白表达量。结果:① 肠道形态学观察:光镜和电镜观察脊髓损伤后黏膜结构破坏明显,肠腔细菌和杂质进入破损的肠黏膜上皮细胞、M细胞和经细胞间连接缝隙进入黏膜下,引起局部炎症反应;经电刺激后均有了很大程度改善。② 锌指蛋白A20:与SG相比,CG各小组A20表达均明显降低(P＜0.01);与SG、CG相比,EG各小组A20的表达均明显升高(P＜0.01)。NOD2:与SG比,CG各组NOD2的表达明显升高(24 h、72 h P＜0.05;48 h P＜0.01);与CG比,EG 48 h、72 h NOD2的表达明显降低( 48 h P＜0.01,72 h P＜0.05);与SG相比,EG各组NOD2表达均无统计学意义。CD68:与SG、EG比,CG各组CD68的表达均明显升高(P＜0.01);与SG相比,EG24 h、48 h组NOD2表达明显升高(P＜0.01),72 h组无统计学意义。结论:电刺激S3神经根能保护肠黏膜机械屏障,减少病原体入侵,改善肠黏膜免疫屏障功能。

关键词: 脊髓损伤, 骶神经电刺激, 肠黏膜免疫屏障

Abstract: Objective: To study the effects of sacral nerve root stimulation on intestinal mucosal immune barrier function in rat with acute complete spinal cord injury(SCI). Methods: Fifty-six Wistar rats were divided into Sham group(SG n=8), control group(CG 24 、 48、 72 h,n=8), and experimental group(EG 24、 48、 72 h,n=8). In CG and EG, according to Fehlings'method,we transected the spinal cord by the aneurysm clip and implanted electrodes into the third sacral foramina on the right side.We stimulated in intensity 4 V,the frequency of 15 Hz,and the pulse of 210 μs.The stimulation period was 2 hours,with 10 minutes stimulation and 10 minutes rest intermittently,twice a day at 8:00-10:00 am and 6:00-8:00 pm. The intestinal morphology was observed under light microscope and electron microscope. The protein expression levels of A20,NOD2,and CD68 by Western blot . Results: ① SCI caused impaired intestinal epithelial barrier function. The intestinal mucosa appeared different degree of damage in CG group; cell-cell connections between intestinal epithelial cells were destroyed; The escherichia coli and other antigen translocated through the injured epithelial cell , M cells, and the leakage to the lamina propria of intestinal villi, which were improved in EG after stimulation.② The expression of A20 in EG was increased ,which had statistical differences between CG or SG(P＜0.01); the expression ofA20 in CG was decreased, which had statistical differences between SG(P＜0.01).The expression of NOD2 in CG was increased, which had statistical differences between SG(24 h,72 h P＜0.05; 48 h P＜0.01);The expression of NOD2 in EG (48 h ,72 h)was decreased, which had statistical differences between CG(48 h P＜0.01,72 h P＜0.05). The expression of NOD2 in EG had no statistical differences between SG. The expression of CD68 in CG was increased,which had statistical differences between SG or EG(P＜0.01).The expression of CD68 in EG was increased in 24 h and 48 h groups ,which had statistical differences between SG(P＜0.01),but had no statistical differences in 72 h group. Conclusion: Sacral nerve root 3 electrostimulation can rehabilitate the peristalsis of intestine,decrease bacterial amount,reduce inflammatory response, enhance endogenous protection, protect the intestinal mucosal immune barrier function.

Key words: spinal cord injury, sacral nerve stimulation, intestinal mucosal immune barrier function

中图分类号:

R454.1

白春宏, 张文丽, 户士忠. 骶神经电刺激对急性脊髓损伤大鼠肠黏膜免疫屏障功能的影响^*[J]. 中国应用生理学杂志, 2020, 36(6): 539-543.

BAI Chun-hong, ZHANG Wen-li, HU Shi-zhong. Sacral nerve stimulating on intestinal mucosal immune barrier function of rats with acute complete spinal cord injury[J]. CJAP, 2020, 36(6): 539-543.

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骶神经电刺激对急性脊髓损伤大鼠肠黏膜免疫屏障功能的影响^*

Sacral nerve stimulating on intestinal mucosal immune barrier function of rats with acute complete spinal cord injury

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