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[1]宋亦祺,張彪,陳陽(yáng),等.扭傷散治療大鼠急性軟組織損傷的效果及作用機(jī)制研究[J].中醫(yī)正骨,2025,37(02):13-20，27.
　SONG Yiqi,ZHANG Biao,CHEN Yang,et al.Efficacy and mechanism of Niushang San(扭傷散)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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扭傷散治療大鼠急性軟組織損傷的效果及作用機(jī)制研究()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第37卷
期數(shù):
2025年02期
頁(yè)碼:
13-20，27
欄目:
基礎(chǔ)研究
出版日期:
2025-02-20

文章信息/Info

Title:
Efficacy and mechanism of Niushang San(扭傷散)in treatment of acute soft tissue injuries in rats:an experimental study
作者:
宋亦祺1; 張彪1; 陳陽(yáng)1; 姜宏2; 俞鵬飛2; 曹峰1
(1.蘇州市中西醫(yī)結(jié)合醫(yī)院,江蘇 蘇州 215101; 2.南京中醫(yī)藥大學(xué)附屬蘇州市中醫(yī)醫(yī)院,江蘇 蘇州 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
關(guān)鍵詞:
軟組織損傷;  巨噬細(xì)胞;  扭傷散;  炎癥;  大鼠;  動(dòng)物實(shí)驗(yàn)
Keywords:
soft tissue injuries;  macrophages;  Niushang San;  inflammation;  rats;  animal experimentation
摘要:
目的:觀察扭傷散治療急性軟組織損傷的效果,并探討其可能的作用機(jī)制。方法:將36只SD大鼠隨機(jī)分為空白組、模型組、扭傷散組和扶他林組,每組9只。空白組大鼠僅脫毛及麻醉,不造模,其余3組大鼠均采用重物打擊股直肌中段構(gòu)建急性軟組織損傷模型。造模后次日開始給藥,扭傷散組外敷扭傷散、扶他林組外涂扶他林(雙氯芬酸二乙胺)乳膠劑,空白組和造模組不進(jìn)行任何處理。記錄干預(yù)1 d、3 d、5 d、7 d后各組大鼠傷肢損傷評(píng)分。末次給藥24 h后,處死大鼠,暴露股直肌中段,切取打擊部位中心的肌肉組織,進(jìn)行組織病理學(xué)觀察。采用實(shí)時(shí)熒光定量逆轉(zhuǎn)錄PCR檢測(cè)肌肉組織中白細(xì)胞介素(interleukin,IL)-1β、腫瘤壞死因子-α(tumor necrosis factor-α,TNF-α)、IL-6的mRNA相對(duì)表達(dá)水平。采用免疫熒光染色法檢測(cè)肌肉組織中誘導(dǎo)型一氧化氮合酶(inducible nitric oxide synthase,iNOS)的蛋白熒光強(qiáng)度。結(jié)果:①大鼠傷肢損傷評(píng)分結(jié)果。模型組、扭傷散組、扶他林組大鼠不同時(shí)間點(diǎn)傷肢損傷評(píng)分隨時(shí)間延長(zhǎng)均呈下降趨勢(shì),但下降趨勢(shì)不完全一致(F=538.942,P=0.000; F=633.208,P=0.000; F=586.374,P=0.000)。干預(yù)1 d后,模型組、扭傷散組、扶他林組大鼠傷肢損傷評(píng)分均高于空白組(P=0.000,P=0.000,P=0.000); 模型組、扭傷散組大鼠傷肢損傷評(píng)分均高于扶他林組(P=0.028,P=0.028); 模型組與扭傷散組大鼠傷肢損傷評(píng)分的差異無統(tǒng)計(jì)學(xué)意義(P=1.000)。干預(yù)3 d、5 d、7 d后,模型組、扭傷散組、扶他林組大鼠傷肢損傷評(píng)分均高于空白組(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); 扭傷散組、扶他林組大鼠傷肢損傷評(píng)分均低于模型組(P=0.002,P=0.001; P=0.001,P=0.005; P=0.015,P=0.005); 扭傷散組與扶他林組大鼠傷肢損傷評(píng)分的差異均無統(tǒng)計(jì)學(xué)意義(P=0.679; P=0.455; P=0.673)。②大鼠損傷肌肉組織病理學(xué)觀察結(jié)果。空白組大鼠肌纖維排列整齊,結(jié)構(gòu)完整,形態(tài)無明顯異常,細(xì)胞核均位于基底膜下。模型組大鼠肌束紊亂,肌纖維扭曲變形,部分?jǐn)嗔选⒆冃詨乃?細(xì)胞核內(nèi)移且大小不均勻,肌間隙不規(guī)則增寬、水腫明顯,可見瘀血或滲出,并有大量炎癥細(xì)胞浸潤(rùn)。扶他林組和扭傷散組大鼠肌纖維的結(jié)構(gòu)和形態(tài)接近正常,僅有部分輕度扭曲,細(xì)胞核位于基底膜下,無明顯細(xì)胞核內(nèi)移現(xiàn)象,核的大小相對(duì)均勻,肌間隙相對(duì)緊密,無明顯水腫,僅有少量炎癥細(xì)胞浸潤(rùn)。③大鼠損傷肌肉組織中IL-1β、TNF-α、IL-6 的mRNA相對(duì)表達(dá)水平檢測(cè)結(jié)果。4組大鼠損傷肌肉組織中IL-1β、TNF-α、IL-6的mRNA相對(duì)表達(dá)水平總體比較,組間差異均有統(tǒng)計(jì)學(xué)意義(F=68.719,P=0.000; F=134.708,P=0.000; F=14.261,P=0.000)。空白組、扭傷散組、扶他林組大鼠損傷肌肉組織中IL-1β、TNF-α、IL-6的mRNA相對(duì)表達(dá)水平均低于模型組(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)。扭傷散組大鼠損傷肌肉組織中IL-1β、TNF-α、IL-6的mRNA相對(duì)表達(dá)水平與空白組的差異均無統(tǒng)計(jì)學(xué)意義(P=0.676,P=0.681,P=0.200)。扶他林組大鼠損傷肌肉組織中IL-1β的mRNA相對(duì)表達(dá)水平高于空白組(P=0.001),TNF-α、IL-6的mRNA相對(duì)表達(dá)水平與空白組的差異均無統(tǒng)計(jì)學(xué)意義(P=0.224,P=0.811)。扭傷散組大鼠損傷肌肉組織中IL-1β的mRNA相對(duì)表達(dá)水平低于扶他林組(P=0.004),TNF-α、IL-6的mRNA相對(duì)表達(dá)水平與扶他林組的差異均無統(tǒng)計(jì)學(xué)意義(P=0.415,P=0.293)。④大鼠損傷肌肉組織中iNOS蛋白熒光強(qiáng)度檢測(cè)結(jié)果。4組大鼠損傷肌肉組織中iNOS蛋白熒光強(qiáng)度總體比較,差異有統(tǒng)計(jì)學(xué)意義(F=420.329,P=0.000)。模型組、扭傷散組、扶他林組大鼠損傷肌肉組織中iNOS蛋白熒光強(qiáng)度均高于空白組(P=0.000,P=0.000,P=0.000)。扭傷散組、扶他林組大鼠損傷肌肉組織中iNOS蛋白熒光強(qiáng)度均低于模型組(P=0.000,P=0.000)。扭傷散組大鼠損傷肌肉組織中iNOS蛋白熒光強(qiáng)度低于扶他林組(P=0.028)。結(jié)論:扭傷散對(duì)大鼠急性軟組織損傷具有顯著的改善作用,且效果與扶他林接近,其作用機(jī)制可能與抑制巨噬細(xì)胞 M1極化,從而減少炎癥因子的釋放有關(guān)。
Abstract:
Objective:To observe the effect of Niushang San(扭傷散,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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備注/Memo

備注/Memo:
基金項(xiàng)目:蘇州市科技發(fā)展計(jì)劃項(xiàng)目(SYSD2020235); 蘇州市應(yīng)用基礎(chǔ)研究(醫(yī)療衛(wèi)生)科技創(chuàng)新項(xiàng)目(SYWD2024331)
通訊作者:曹峰 E-mail:[email protected]
更新日期/Last Update: 1900-01-01