中医正骨2025，Vol.37Issue(2)：13-20,27,9.
扭伤散治疗大鼠急性软组织损伤的效果及作用机制研究
Efficacy and mechanism of Niushang San(扭伤散)in treatment of acute soft tissue injuries in rats:an experi-mental study
摘要
Abstract
Objective:To observe the effect of Niushang San(扭伤散,NSS)in treatment of acute soft tissue injuries in rats,and to ex-plore 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 femo-ris muscle with a heavy object for inducing acute soft tissue injuries,while the ones in blank group merely underwent hair removal and anes-thesia.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 high-er 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,ad-ditionally,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 in-flammatory 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 tis-sues 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 ex-pression 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 in-flammatory factors.关键词
软组织损伤/巨噬细胞/扭伤散/炎症/大鼠/动物实验Key words
soft tissue injuries/macrophages/Niushang San/inflammation/rats/animal experimentation引用本文复制引用
宋亦祺,张彪,陈阳,沈为苟,姜宏,俞鹏飞,曹峰..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(2):13-20,27,9.基金项目
苏州市科技发展计划项目(SYSD2020235) （SYSD2020235）
苏州市应用基础研究(医疗卫生)科技创新项目(SYWD2024331) （医疗卫生）
