| 吴静泽,王晔恺,陈位,金丹雯.昆仙胶囊联合吲哚美辛通过抑制BMP4/Smad5信号通路治疗创伤性骨化性肌炎大鼠的实验研究[J].浙江中西医结合杂志,2024,34(8): |
| 昆仙胶囊联合吲哚美辛通过抑制BMP4/Smad5信号通路治疗创伤性骨化性肌炎大鼠的实验研究 |
| Effect of kunxian capsule and Indomethacin on rat model with traumaticSmyositisSossificans by inhibition BMP4/Smad5 signal pathway |
| 投稿时间：2023-12-12 修订日期：2024-07-22 |
| DOI： |
| 中文关键词: 大鼠 骨化性肌炎 昆仙胶囊 吲哚美辛 骨成型蛋白4 Smad蛋白 |
| 英文关键词:Myositis ossificans Kunxian capsule Indomethacin |
| 基金项目:浙江省中医药科技计划项目（No.2020ZB274）；浙江省医药卫生科技计划项目（No.2020KY340） |
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| 中文摘要: |
| 目的 探讨昆仙胶囊联合吲哚美辛治疗创伤性骨化性肌炎模型大鼠的可能作用机制。方法 96只雄性SD大鼠通过切断跟腱和破坏踝关节建立骨化性肌炎模型，按随机数字表法分为对照组、中药组、西药组、中西药联合组，每组24只；术后当天，对照组、中药组、西药组、中西药联合组分别给予0.2 g/50 mL安慰剂、0.3 g/50 mL昆仙胶囊粉剂溶液、0.1 g/50 mL吲哚美辛粉剂溶液、0.3 g/50 mL昆仙胶囊粉剂和0.1 g/50 mL吲哚美辛粉剂混合溶液进行灌胃，每天1次，每次2 mL，时间2~8周。在术后2、4、6、8周，每组各断颈处死6只大鼠，用X光机对大鼠踝关节进行摄片,测量踝关节肿胀厚度，取血清用酶联免疫吸附试验（ELISA）检测转化生长因子-β（TGF-β）水平，取跟腱组织用免疫组化检测骨成型蛋白4（BMP4）和Smad5蛋白表达。结果 在踝关节肿胀厚度中，与对照组同一时间点比较，4周的中西药联合组降低 [（10.77±0.88） mm比（13.78±0.91） mm，P＜0.05]，6周的中药组、西药组、中西药联合组降低 [（11.82±1.08） mm、（11.34±0.88） mm、（8.97±0.53） mm比（12.97±0.51） mm，P均＜0.05]、8周的中药组、西药组、中西药联合组降低[（9.39±0.90） mm、（9.47±0.45） mm、（8.47±0.38） mm比（12.25±0.89） mm，P均＜0.05]。
血清TGF-β浓度中，与对照组同一时间点比较，2周的西药组和中西药联合组降低[（386.00±29.46） ng/L、（376.50±27.96） ng/L、ng/L比（484.50±59.49） ng/L，P均＜0.05]
4周的中药组、西药组、中西药联合组降低 [（374.33±31.48） ng/L、（331.83±47.11） ng/L、（337.66±46.86） ng/L比（453.16±53.11） ng/L，P均＜0.05]、6周的中药组、西药组、中西药联合组降低[（346.66±47.84） ng/L、（304.50±26.25） ng/L、（277.33±44.79） ng/L比（417.00±32.58） ng/L，P均＜0.05]、8周的中药组、西药组、中西药联合组降低[（307.16±24.46） ng/L、（283.33±34.85） ng/L、（169.50±31.64） ng/L比（384.83±42.22） ng/L，P均＜0.05]。在跟腱组织BMP4平均光密度（AOD）中，与对照组同一时间点比较，4周的中药组和中西药联合组 [（148.18±15.88）、（146.01±14.33）比（189.41±14.40），P均＜0.05]、6周的中药组、西药组和中西药联合组 [（137.00±8.95）、（123.15±22.93）、（112.53±35.93）比（172.98±7.56），P均＜0.05]、8周的中药组、西药组和中西药联合组[（128.21±12.85）、（117.58±21.34）、（96.04±21.33）比（157.95±10.41），P均＜0.05]均降低。在跟腱组织Smad5 AOD中，与对照组同一时间点相比，2周的中西药联合组降低[（101.98±9.46）比（130.94±9.54），P均＜0.05]，4周的中药组、西药组和中西药联合组降低 [（92.65±12.96）、（96.97±12.85）、（93.61±11.44）比（121.01±10.23），P均＜0.05]、6周的中药组、西药组和中西药联合组降低[（85.23±12.73）、（87.73±13.07）、（76.96±12.71）比（114.18±10.29），P均＜0.05]、8周的中药组和中西药联合组降低 [（79.91±13.19）、（76.17±21.18）比（94.74±19.93），P均＜0.05]。结论 昆仙胶囊联合吲哚美辛对创伤性骨化性肌炎的抑制效果有增益作用，其机理可能和调控BMP4/Smad5信号通路有关。 |
| 英文摘要: |
| Objective To invesigate the combined effect of kunxian capsule and indomethacin in a rat model of traumatic myositis ossificans. Methods A totle of 96 SD rats were treated by cutting the achilles tendon and damaging the ankle joint. Then they were divided into 4 groups:control group(24 rats), traditional Chinese medicine group(24 rats), western medicine group(24 rats) and combination of traditional Chinese and western medicine group(24 rats).From the day of operation, control group, traditional Chinese medicine group group, western medicine group and combination group were given 0.2g/50ml placebo solution, 0.3g/50ml kunxian capsule solution, 0.1g/50ml indomethacin solution, 0.3g/50ml kunxian capsule and 0.1g/50ml indomethacin mixed solution respectively, gavage administration with 2ml once a day. After operation, 6 rats were killed by cervical dislocations at 2 weeks, 4 weeks, 6 weeks, 8 weeks, respectively. The swelling thickness of ankle joint was examined by X-ray and measured by vernier caliper. Serum levels of TGF-β were detected by enzyme linked immunosorbent assay(ELISA). BMP4 and Smad5 protein expression in achilles tendon tissue were detected by immunohistochemistry. Results In swelling thickness, compared to control group at the same time, combination group at 4 week decreased(10.77±0.88 vs. 13.78±0.91 mm; P<0.05); traditional Chinese medicine group, western medicine group and combination group at 6 week decreased(11.82±1.08,11.34±0.88 and 8.97±0.53 vs.12.97±0.51 mm;P<0.05); traditional Chinese medicine group, western medicine group and combination group at 8 week decreased(9.39± 0.90, 9.47±0.45 and 8.47±0.38 vs. 12.25±0.89 mm,P<0.05). In serum levels of TGF-β, compared to control group at the same time, western medicine group and combination group at 2 week decreased(386.00±29.46, 376.50±27.96 vs. 484.50±59.49 ng/L;P<0.05);traditional Chinese medicine group, western medicine group and combination group at 4 week decreased(374.33±31.48, 331.83±47.11 and 337.66± 46.86 vs. 453.16±53.11 ng/L;P<0.05); traditional Chinese medicine group, western medicine group and combination group at 6 week decreased (346.66±47.84, 304.50±26.25 and 277.33± 44.79 vs. 417.00±32.58 ng/L;P<0.05); traditional Chinese medicine group, western medicine group and combination group at 8 week decreased(307.16±24.46, 283.33 ±34.85 and 169.50±31.64 vs. 384.83±42.22 ng/L;P<0.05). In BMP4 expression of achilles tendon tissue, compared to control group at same time, traditional Chinese medicine group and combination group at 4 week decreased(148.18±15.88, 146.01±14.33 vs. 189.41±14.40;P<0.05); traditional Chinese medicine group, western medicine group and combination group at 6 week decreased(137.00±8.95, 123.15±22.93, 112.53±35.93 vs. 172.98±7.56;P<0.05); traditional Chinese medicine group, western medicine group and combination group at 8 week decreased(128.21±12.85, 117.58±21.34, 96.04±21.33 vs. 157.95±10.41;P<0.05).In Smad5 expressin of achilles tendon tissue, compared to control group at the same time, combination group at 2 week decreased(101.98±9.46 vs. 130.94±9.54;P<0.05);traditional Chinese medicine group, western medicine group and combination group at 4 week decreased(92.65±12.96, 96.97±12.85, 93.61±11.44 vs. 121.01±10.23;P<0.05); traditional Chinese medicine group, western medicine group and combination group at 6 week decreased(85.23±12.73, 87.73±13.07, 76.96±12.71 vs. 114.18±10.29;P<0.05); traditional Chinese medicine group and combination group at 8 week decreased(79.91±13.19, 76.17±21.18 vs. 94.74±19.93;P<0.05). Conclusion Combined effect of Kunxian capsule and indomethacin on traumatic myositis ossificans by mechanism of inhibiting BMP4/Smad5 signal pathway. |
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