[1]宋亦祺,张彪,陈阳,等.扭伤散治疗大鼠急性软组织损伤的效果及作用机制研究[J].中医正骨,2025,37(02):13-20，27.
　SONG Yiqi,ZHANG Biao,CHEN Yang,et al.Efficacy and mechanism of Niushang San(扭伤散)in treatment of acute soft tissue injuries in rats:an experimental study[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2025,37(02):13-20，27.
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扭伤散治疗大鼠急性软组织损伤的效果及作用机制研究()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第37卷
期数:
2025年02期
页码:
13-20，27
栏目:
基础研究
出版日期:
2025-02-20

文章信息/Info

Title:
Efficacy and mechanism of Niushang San(扭伤散)in treatment of acute soft tissue injuries in rats:an experimental study
作者:
宋亦祺1; 张彪1; 陈阳1; 沈为苟1; 姜宏2; 俞鹏飞2; 曹峰1
(1.苏州市中西医结合医院,江苏 苏州 215101; 2.南京中医药大学附属苏州市中医医院,江苏 苏州 215009)
Author(s):
SONG Yiqi1; ZHANG Biao1; CHEN Yang1; SHEN Weigou1; JIANG Hong2; YU Pengfei2; CAO Feng1
1.Suzhou Hospital of Integrated Traditional Chinese and Western Medicine,Suzhou 215101,Jiangsu,China 2.Suzhou Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Chinese Medicine,Suzhou 215009,Jiangsu,China
关键词:
软组织损伤;  巨噬细胞;  扭伤散;  炎症;  大鼠;  动物实验
Keywords:
soft tissue injuries;  macrophages;  Niushang San;  inflammation;  rats;  animal experimentation
摘要:
目的:观察扭伤散治疗急性软组织损伤的效果,并探讨其可能的作用机制。方法:将36只SD大鼠随机分为空白组、模型组、扭伤散组和扶他林组,每组9只。空白组大鼠仅脱毛及麻醉,不造模,其余3组大鼠均采用重物打击股直肌中段构建急性软组织损伤模型。造模后次日开始给药,扭伤散组外敷扭伤散、扶他林组外涂扶他林(双氯芬酸二乙胺)乳胶剂,空白组和造模组不进行任何处理。记录干预1 d、3 d、5 d、7 d后各组大鼠伤肢损伤评分。末次给药24 h后,处死大鼠,暴露股直肌中段,切取打击部位中心的肌肉组织,进行组织病理学观察。采用实时荧光定量逆转录PCR检测肌肉组织中白细胞介素(interleukin,IL)-1β、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、IL-6的mRNA相对表达水平。采用免疫荧光染色法检测肌肉组织中诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)的蛋白荧光强度。结果:①大鼠伤肢损伤评分结果。模型组、扭伤散组、扶他林组大鼠不同时间点伤肢损伤评分随时间延长均呈下降趋势,但下降趋势不完全一致(F=538.942,P=0.000; F=633.208,P=0.000; F=586.374,P=0.000)。干预1 d后,模型组、扭伤散组、扶他林组大鼠伤肢损伤评分均高于空白组(P=0.000,P=0.000,P=0.000); 模型组、扭伤散组大鼠伤肢损伤评分均高于扶他林组(P=0.028,P=0.028); 模型组与扭伤散组大鼠伤肢损伤评分的差异无统计学意义(P=1.000)。干预3 d、5 d、7 d后,模型组、扭伤散组、扶他林组大鼠伤肢损伤评分均高于空白组(P=0.000,P=0.000,P=0.000; P=0.000,P=0.000,P=0.000; P=0.000,P=0.005,P=0.015); 扭伤散组、扶他林组大鼠伤肢损伤评分均低于模型组(P=0.002,P=0.001; P=0.001,P=0.005; P=0.015,P=0.005); 扭伤散组与扶他林组大鼠伤肢损伤评分的差异均无统计学意义(P=0.679; P=0.455; P=0.673)。②大鼠损伤肌肉组织病理学观察结果。空白组大鼠肌纤维排列整齐,结构完整,形态无明显异常,细胞核均位于基底膜下。模型组大鼠肌束紊乱,肌纤维扭曲变形,部分断裂、变性坏死,细胞核内移且大小不均匀,肌间隙不规则增宽、水肿明显,可见瘀血或渗出,并有大量炎症细胞浸润。扶他林组和扭伤散组大鼠肌纤维的结构和形态接近正常,仅有部分轻度扭曲,细胞核位于基底膜下,无明显细胞核内移现象,核的大小相对均匀,肌间隙相对紧密,无明显水肿,仅有少量炎症细胞浸润。③大鼠损伤肌肉组织中IL-1β、TNF-α、IL-6 的mRNA相对表达水平检测结果。4组大鼠损伤肌肉组织中IL-1β、TNF-α、IL-6的mRNA相对表达水平总体比较,组间差异均有统计学意义(F=68.719,P=0.000; F=134.708,P=0.000; F=14.261,P=0.000)。空白组、扭伤散组、扶他林组大鼠损伤肌肉组织中IL-1β、TNF-α、IL-6的mRNA相对表达水平均低于模型组(P=0.000,P=0.000,P=0.000; P=0.000,P=0.000,P=0.000; P=0.000,P=0.000,P=0.000)。扭伤散组大鼠损伤肌肉组织中IL-1β、TNF-α、IL-6的mRNA相对表达水平与空白组的差异均无统计学意义(P=0.676,P=0.681,P=0.200)。扶他林组大鼠损伤肌肉组织中IL-1β的mRNA相对表达水平高于空白组(P=0.001),TNF-α、IL-6的mRNA相对表达水平与空白组的差异均无统计学意义(P=0.224,P=0.811)。扭伤散组大鼠损伤肌肉组织中IL-1β的mRNA相对表达水平低于扶他林组(P=0.004),TNF-α、IL-6的mRNA相对表达水平与扶他林组的差异均无统计学意义(P=0.415,P=0.293)。④大鼠损伤肌肉组织中iNOS蛋白荧光强度检测结果。4组大鼠损伤肌肉组织中iNOS蛋白荧光强度总体比较,差异有统计学意义(F=420.329,P=0.000)。模型组、扭伤散组、扶他林组大鼠损伤肌肉组织中iNOS蛋白荧光强度均高于空白组(P=0.000,P=0.000,P=0.000)。扭伤散组、扶他林组大鼠损伤肌肉组织中iNOS蛋白荧光强度均低于模型组(P=0.000,P=0.000)。扭伤散组大鼠损伤肌肉组织中iNOS蛋白荧光强度低于扶他林组(P=0.028)。结论:扭伤散对大鼠急性软组织损伤具有显著的改善作用,且效果与扶他林接近,其作用机制可能与抑制巨噬细胞 M1极化,从而减少炎症因子的释放有关。
Abstract:
Objective:To observe the effect of Niushang San(扭伤散,NSS)in treatment of acute soft tissue injuries in rats,and to explore its underlying mechanism.Methods:Thirty-six SD rats were selected and randomized into blank group,model group,NSS group and voltaren group,with 9 ones in each group.All rats but the ones in blank group were modeled by striking the mid-segment of the rectus femoris muscle with a heavy object for inducing acute soft tissue injuries,while the ones in blank group merely underwent hair removal and anesthesia.The next day after successful modeling,the rats in NSS group and voltaren group were treated with external application of NSS and voltaren(diclofenac diethylamine)emulgel,respectively,while the ones in blank group and model group were not given any treatment.The injury severity score of the injured limbs in each group were assessed and recorded after 1-,3-,5- and 7-day intervention,respectively.Twenty-four hours after the last medication,the rats were sacrificed and the mid-segment of their rectus femoris muscle were exposed.The muscle tissues in the center of the striked site were harvested and stained with hematoxylin-eosin(HE)for observing the histopathological changes.Furthermore,the relative mRNA expression levels of interleukin(IL)-1β,tumor necrosis factor-α(TNF-α)and IL-6 in the injured muscle tissues were detected by real-time fluorescence quantitative reverse transcription PCR(fqRT-PCR),and the fluorescence intensity of inducible nitric oxide synthase(iNOS)protein in the injured muscle tissues was detected by immunofluorescence staining.Results:①The injury severity score of the injured limbs of rats.The injury severity score of the injured limbs presented a downward trend over time in mo-del group,NSS group and voltaren group,while the downward trend was inconsistent with each other in the 3 groups(F=538.942,P=0.000; F=633.208,P=0.000; F=586.374,P=0.000).After 1-day intervention,the injury severity score of the injured limbs was higher in model group,NSS group and voltaren group compared to blank group(P=0.000,P=0.000,P=0.000),and was higher in model group and NSS group compared to voltaren group(P=0.028,P=0.028),while,it was not significantly different from each other between model group and NSS group(P=1.000).After 3-,5- and 7-day intervention,the injury severity score of the injured limbs was higher in model group,NSS group and voltaren group compared to blank group(P=0.000,P=0.000,P=0.000; P=0.000,P=0.000,P=0.000; P=0.000,P=0.005,P=0.015),and was higher in model group compared to NSS group and voltaren group(P=0.002,P=0.001; P=0.001,P=0.005; P=0.015,P=0.005),while,it was not significantly different from each other between NSS group and voltaren group(P=0.679; P=0.455; P=0.673).②The histopathological changes in the injured muscle tissues of rats.The HE staining results showed that the muscle fibers of the rats in the blank group were neatly arranged with intact structure and no obvious abnormaity in morphology,additionally,the cell nuclei were all located beneath the basement membrane.In the model group,the rat muscle bundles were disordered,the muscle fibers were twisted and deformed,with some fractured,denatured,and necrotic.The cell nuclei shifted inward with uneven sizes,the muscle gaps were irregularly widened with obvious edema,besides,the stagnated blood or exudation,accompanied by a large number of inflammatory cell infiltration,were observed.The structure and morphology of muscle fibers in rats of votalin group and NSS group were close to normal,with only some slightly twisted.The cell nuclei located beneath the basement membrane without obvious inward shifting,and the size of the nuclei was relatively uniform; besides,the muscle gaps were relatively tight without significant edema,moreover,a small amount of inflammatory cell infiltration were observed.③The relative mRNA expression levels of IL-1β,TNF-α and IL-6 in the injured muscle tissues of rats.There was statistical difference in the relative mRNA expression levels of IL-1β,TNF-α and IL-6 in the injured muscle tissues of rats among the 4 groups in general(F=68.719,P=0.000; F=134.708,P=0.000; F=14.261,P=0.000).The relative mRNA expression levels of IL-1β,TNF-α and IL-6 in the injured muscle tissues of rats were lower in blank group,NSS group and voltaren group compared to model group(P=0.000,P=0.000,P=0.000; P=0.000,P=0.000,P=0.000; P=0.000,P=0.000,P=0.000),while,they were not significantly different from each other between NSS group and blank group(P=0.676,P=0.681,P=0.200).The relative mRNA expression level of IL-1β in the injured muscle tissues of rats was higher in voltaren group compared to blank group(P=0.001),while those of TNF-α and IL-6 were not significantly different from each other between the 2 groups(P=0.224,P=0.811).The relative mRNA expression level of IL-1β in the injured muscle tissues of rats was lower in NSS group compared to voltaren group(P=0.004),while those of TNF-α and IL-6 were not significantly different from each other between the 2 groups(P=0.415,P=0.293).④The fluorescence intensity of iNOS protein in the injured muscle tissues of rats.There was statistical difference in the fluorescence intensity of iNOS protein in the injured muscle tissues of rats among the 4 groups in general(F=420.329,P=0.000).The fluorescence intensity of iNOS protein in the injured muscle tissues of rats was higher in model group,NSS group and voltaren group compared to blank group(P=0.000,P=0.000,P=0.000),and was higher in model group compared to NSS group and voltaren group(P=0.000,P=0.000),and was higher in voltaren group compared to NSS group(P=0.028).Conclusion:NSS has a significant effect on improving acute soft tissue injuries in rats,and its efficacy is similar to that of voltaren.It may work by inhibiting the polarization of M1 macrophage,thereby reducing the release of inflammatory factors.

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备注/Memo

备注/Memo:
基金项目:苏州市科技发展计划项目(SYSD2020235); 苏州市应用基础研究(医疗卫生)科技创新项目(SYWD2024331)
通讯作者:曹峰 E-mail:szmdcf@139.com
更新日期/Last Update: 1900-01-01