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[1]廖瑩瑩,張鑫,任凱,等.鄭氏芪藤軟堅散外敷治療創(chuàng)傷性膝關(guān)節(jié)僵硬的實驗研究[J].中醫(yī)正骨,2023,35(02):1-9,21.
　LIAO Yingying,ZHANG Xin,REN Kai,et al.An experimental study of external application of Zheng's Qiteng Ruanjian San in the treatment of post-traumatic knee joint stiffness[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2023,35(02):1-9,21.
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鄭氏芪藤軟堅散外敷治療創(chuàng)傷性膝關(guān)節(jié)僵硬的實驗研究()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第35卷
期數(shù):
2023年02期
頁碼:
1-9,21
欄目:
基礎(chǔ)研究
出版日期:
2023-02-20

文章信息/Info

Title:
An experimental study of external application of Zheng's Qiteng Ruanjian San in the treatment of post-traumatic knee joint stiffness
作者:
廖瑩瑩1; 張鑫1; 任凱1; 劉輝1; 陳科1; 溫呈洪2; 稅曉平3
(1.四川省骨科醫(yī)院,四川 成都 610041; 2.成都體育學(xué)院附屬體育醫(yī)院,四川 成都 610041; 3.綿陽市骨科醫(yī)院,四川 綿陽 621000)
Author(s):
LIAO Yingying1; ZHANG Xin1; REN Kai1; LIU Hui1; CHEN Ke1; WEN Chenghong2; SHUI Xiaoping3
1.Sichuan Province Orthopedic Hospital,Chengdu 610041,Sichuan,China 2.Affiliated Sport Hospital of Chengdu Sport University,Chengdu 610041,Sichuan,China 3.Mianyang Orthopedics Hospital,Mianyang 621000,Sichuan,China
關(guān)鍵詞:
膝關(guān)節(jié);  關(guān)節(jié)僵硬;  創(chuàng)傷和損傷;  鄭氏芪藤軟堅散;  動物實驗
Keywords:
knee joint;  joint stiffness;  wounds and injuries;  Zheng's Qiteng Ruanjian San;  animal experimentation
摘要:
目的:觀察鄭氏芪藤軟堅散外敷治療創(chuàng)傷性膝關(guān)節(jié)僵硬的療效,并探討其可能的作用機制。方法:從66只8周齡SPF級雄性SD大鼠中隨機選取18只納入空白組,剩余48只建立右側(cè)創(chuàng)傷性膝關(guān)節(jié)屈曲型僵硬模型,將造模成功的41只大鼠隨機納入模型組(20只)和軟堅散組(21只)。空白組和模型組大鼠常規(guī)喂養(yǎng),不予干預(yù); 軟堅散組大鼠造模成功后采用鄭氏芪藤軟堅散外敷,每天1次,連續(xù)治療4周。分別于藥物干預(yù)開始前、藥物干預(yù)開始后2周、藥物干預(yù)結(jié)束后,從各組隨機選取部分大鼠,分別進行右側(cè)膝關(guān)節(jié)活動度測量、膝關(guān)節(jié)囊厚度測量、膝關(guān)節(jié)囊組織病理學(xué)觀察及成纖維細(xì)胞計數(shù)(HE染色)、膝關(guān)節(jié)囊中轉(zhuǎn)化生長因子β1(transforming growth factor β1,TGF-β1)水平測定(免疫組織化學(xué)法)。結(jié)果:①膝關(guān)節(jié)活動度。在3個時間點,3組大鼠的膝關(guān)節(jié)活動度總體比較,差異均有統(tǒng)計學(xué)意義(122.43°±4.54°,64.78°±2.68°,65.13°±4.52°,F=530.817,P=0.000; 124.50°±5.01°,82.83°±6.05°,88.67°±8.62°,F=67.404,P=0.000; 122.33°±4.08°,92.50°±2.74°,107.13°±7.41°,F=44.886,P=0.000)。在3個時間點,空白組的膝關(guān)節(jié)活動度均大于模型組和軟堅散組(P=0.000,P=0.000; P=0.000,P=0.000; P=0.000,P=0.000); 藥物干預(yù)開始前、藥物干預(yù)開始后2周時,模型組和軟堅散組的膝關(guān)節(jié)活動度比較,組間差異均無統(tǒng)計學(xué)意義(P=0.857,P=0.154); 藥物干預(yù)結(jié)束后,軟堅散組的膝關(guān)節(jié)活動度大于模型組(P=0.000)。②膝關(guān)節(jié)囊厚度。在3個時間點,3組大鼠的膝關(guān)節(jié)囊厚度總體比較,差異均有統(tǒng)計學(xué)意義[(0.256±0.020)mm,(0.533±0.024)mm,(0.528±0.012)mm,F=412.282,P=0.000;(0.254±0.036)mm,(0.531±0.018)mm,(0.523±0.019)mm,F=125.803,P=0.000;(0.257±0.028)mm,(0.527±0.017)mm,(0.521±0.014)mm,F=211.708,P=0.000]。在3個時間點,空白組的膝關(guān)節(jié)囊厚度均小于模型組(P=0.000,P=0.000,P=0.000); 藥物干預(yù)開始前和藥物干預(yù)結(jié)束后,軟堅散組的膝關(guān)節(jié)囊厚度均大于空白組(P=0.000,P=0.000); 藥物干預(yù)開始后2周時,軟堅散組與空白組的膝關(guān)節(jié)囊厚度的差異無統(tǒng)計學(xué)意義(P=0.053); 在3個時間點,模型組與軟堅散組的膝關(guān)節(jié)囊厚度比較,組間差異均無統(tǒng)計學(xué)意義(P=0.702,P=0.740,P=0.725)。③膝關(guān)節(jié)囊組織成纖維細(xì)胞計數(shù)結(jié)果。在3個時間點,3組大鼠膝關(guān)節(jié)囊中成纖維細(xì)胞數(shù)量總體比較,差異均有統(tǒng)計學(xué)意義[(1.67±0.98)個,(98.25±17.97)個,(103.92±11.85)個,F=85.392,P=0.000;(1.93±1.09)個,(68.92±17.50)個,(46.17±7.37)個,F=48.565,P=0.000;(1.80±0.96)個,(49.83±11.41)個,(31.73±10.66)個,F=34.755,P=0.000]。在3個時間點,空白組膝關(guān)節(jié)囊中成纖維細(xì)胞數(shù)量均少于模型組和軟堅散組(P=0.000,P=0.000; P=0.000,P=0.000; P=0.000,P=0.000); 藥物干預(yù)開始前,軟堅散組和模型組膝關(guān)節(jié)囊中成纖維細(xì)胞數(shù)量比較,差異無統(tǒng)計學(xué)意義(P=0.535); 藥物干預(yù)開始后2周和藥物干預(yù)結(jié)束后,軟堅散組膝關(guān)節(jié)囊中成纖維細(xì)胞數(shù)量均少于模型組(P=0.011,P=0.010)。④膝關(guān)節(jié)囊中TGF-β1水平。在3個時間點,3組大鼠膝關(guān)節(jié)囊中TGF-β1水平總體比較,差異均有統(tǒng)計學(xué)意義[(0.148±0.060)%,(2.454±1.392)%,(2.519±0.653)%,F=9.240,P=0.007;(0.147±0.054)%,(1.136±0.491)%,(0.271±0.095)%,F=13.765,P=0.002;(0.147±0.059)%,(0.765±0.339)%,(0.190±0.070)%,F=13.105,P=0.002]。藥物干預(yù)開始前,模型組和軟堅散組膝關(guān)節(jié)囊中TGF-β1水平均高于空白組(P=0.005,P=0.004),軟堅散組與模型組膝關(guān)節(jié)囊中TGF-β1水平的差異無統(tǒng)計學(xué)意義(P=0.920); 藥物干預(yù)開始后2周時,模型組膝關(guān)節(jié)囊中TGF-β1水平高于空白組和軟堅散組(P=0.001,P=0.002),軟堅散組與空白組膝關(guān)節(jié)囊中TGF-β1水平的差異無統(tǒng)計學(xué)意義(P=0.563); 藥物干預(yù)結(jié)束后,模型組膝關(guān)節(jié)囊中TGF-β1水平高于空白組和軟堅散組(P=0.001,P=0.001),軟堅散組與空白組膝關(guān)節(jié)囊中TGF-β1水平的差異無統(tǒng)計學(xué)意義(P=0.745)。結(jié)論:早期應(yīng)用鄭氏芪藤軟堅散,可以促進創(chuàng)傷性膝關(guān)節(jié)僵硬模型大鼠膝關(guān)節(jié)功恢復(fù),抑制成纖維細(xì)胞增殖和TGF-β1表達可能是其作用機制之一。
Abstract:
Objective:To observe the effect and underlying mechanism of external application of Zheng's Qiteng Ruanjian San in the treatment of post-traumatic knee joint stiffness.Methods:Sixty-six 8-week-old male SD rats of SPF grade were enrolled and randomly divided into a blank group(n=18)and an experimental group(n=48).The right post-traumatic knee joint stiffness model of flexion type was induced in 48 rats of the experimental group.Forty-one successful model rats were randomly divided into a model group(n=20)and a Qiteng Ruanjian San group(n=21).Rats in the blank group and the model group were fed routinely without intervention,while those in the Qiteng Ruanjian San group received external application of Qiteng Ruanjian San,once a day for 4 weeks after modeling.Some rats were randomly selected from each group before drug intervention,at 2 weeks of drug intervention,and after drug intervention for the measurement of right knee joint range of motion and knee capsule thickness,histopathological observation of knee capsule,fibroblast count(HE staining),and the determination of transforming growth factor β1(TGF-β1)level in the knee capsule(immunohistochemistry).Results:①Knee joint range of motion.There were statistically significant differences in the overall knee joint range of motion among the blank group,the model group,and the Qiteng Ruanjian San group at three time points(122.43°±4.54° vs 64.78°±2.68° vs 65.13°±4.52°,F=530.817,P=0.000; 124.50°±5.01° vs 82.83°±6.05° vs 88.67°±8.62°,F=67.404,P=0.000; 122.33°±4.08° vs 92.50°±2.74° vs 107.13°±7.41°,F=44.886,P=0.000).At the three time points,the knee joint range of motion in the blank group was greater than those in the model group and the Qiteng Ruanjian San group(P=0.000,P=0.000; P=0.000,P=0.000; P=0.000,P=0.000).There was no significant difference in the knee joint range of motion between the model group and the Qiteng Ruanjian San group before drug intervention and at 2 weeks of drug intervention(P=0.857,P=0.154),and the knee joint range of motion in the Qiteng Ruanjian San group was greater than that in the model group after drug intervention(P=0.000).②Knee capsule thickness.There were overall statistically significant differences in the knee joint capsule thickness among the blank group,the model group,and the Qiteng Ruanjian San group at three time points(0.256±0.020 vs 0.533±0.024 vs 0.528±0.012 mm,F=412.282,P=0.000; 0.254±0.036 vs 0.531±0.018 vs 0.523±0.019 mm,F=125.803,P=0.000; 0.257±0.028 vs 0.527±0.017 vs 0.521±0.014 mm,F=211.708,P=0.000).At the three time points,the knee capsule thickness of the blank group was smaller than that of the model group(P=0.000,P=0.000,P=0.000),and the knee capsule thickness of the Qiteng Ruanjian San group was greater than that of the blank group before and after drug intervention(P=0.000,P=0.000).There was no significant difference in the knee capsule thickness between the Qiteng Ruanjian San group and the blank group at 2 weeks of drug intervention(P=0.053).At the three time points,there was no significant difference in the knee capsule thickness between the model group and the Qiteng Ruanjian San group(P=0.702,P=0.740,P=0.725).③Fibroblast count results of knee capsule tissues.There were statistically significant differences in fibroblast count of knee capsule tissues among the blank group,the model group,and the Qiteng Ruanjian San group at three time points(1.67±0.98 vs 98.25±17.97 vs 103.92±11.85,F=85.392,P=0.000; 1.93±1.09 vs 68.92±17.50 vs 46.17±7.37,F=48.565,P=0.000; 1.80±0.96 vs 49.83±11.41 vs 31.73±10.66,F=34.755,P=0.000).At the three time points,the fibroblast count in knee capsule tissues of the blank group was less than those of the model group and the Qiteng Ruanjian San groups(P=0.000,P=0.000; P=0.000,P=0.000; P=0.000,P=0.000).There was no significant difference in the fibroblast count in knee capsule tissues between the Qiteng Ruanjian San group and the model group before drug intervention(P=0.535),and the fibroblast count in knee capsule tissues of the Qiteng Ruanjian San group was less than that of the model group at 2 weeks of drug intervention and after drug intervention(P=0.011,P=0.010).④TGF-β1 levels in the knee capsule.There were statistically significant differences in TGF-β1 levels among the blank group,the model group,and the Qiteng Ruanjian San group at three time points(0.148±0.060 vs 2.454±1.392 vs 2.519±0.653%,F=9.240,P=0.007; 0.147±0.054 vs 1.136±0.491 vs 0.271±0.095%,F=13.765,P=0.002; 0.147±0.059 vs 0.765±0.339 vs 0.190±0.070%,F=13.105,P=0.002).Before drug intervention,the TGF-β1 levels in the knee capsule of the model group and the Qiteng Ruanjian San group were higher than that of the blank group(P=0.005,P=0.004),and there was no significant difference in the TGF-β1 levels in the knee capsule between the Qiteng Ruanjian San group and the model group(P=0.920).At 2 weeks of drug intervention,the TGF-β1 level in the knee capsule of the model group was higher than those of the blank group and the Qiteng Ruanjian San group(P=0.001,P=0.002),and there was no significant difference in the TGF-β1 levels in the knee capsule between the Qiteng Ruanjian San group and the blank group(P=0.563).After drug intervention,the TGF-β1 level in the knee capsule of the model group was higher than those of the blank group and the Qiteng Ruanjian San group(P=0.001,P=0.001),and there was no significant difference in the TGF-β1 levels in the knee capsule between the Qiteng Ruanjian San group and the blank group(P=0.745).Conclusion:Early application of Zheng's Qiteng Ruanjian San can improve the knee joint function of rats with post-traumatic knee joint stiffness,and the inhibition of fibroblast proliferation and TGF-β1 expression may be one of its mechanisms.

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(收稿日期:2022-08-25 本文編輯:李曉樂)

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備注/Memo

備注/Memo:
基金項目:四川省中醫(yī)藥管理局科學(xué)技術(shù)研究專項課題(2020JC0098);四川省區(qū)域中醫(yī)(專科)診療中心建設(shè)項目(川中醫(yī)藥函〔2018〕20號) 通訊作者:張鑫 E-mail:[email protected]
更新日期/Last Update: 1900-01-01