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、0.3g/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,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 与对照组同一时间点比较,中西药联合组4周时[(10.77±0.88)mm比(13.78±0.91)mm,P<0.05],中药组、西药组、中西药联合组6周[(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周时[(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]踝关节肿胀厚度均降低。与对照组同一时间点比较,中药组、西药组、中西药联合组4周[(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[(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[(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]血清TGF-β浓度均降低。与对照组同一时间点比较,中药组和中西药联合组4周时[(148.18±15.88)、(146.01±14.33)比(189.41±14.40),P均<0.05]、中药组、西药组和中西药联合组6周[(137.00±8.95)、(123.15±22.93)、(112.53±35.93)比(172.98±7.56),P均<0.05]、8周时[(128.21±12.85)、(117.58±21.34)、(96.04±21.33)比(157.95±10.41),P均<0.05]跟腱组织BMP4平均光密度(AOD)均降低。与对照组同一时间点相比,中药组、西药组和中西药联合组4周[(92.65±12.96)、(96.97±12.85)、(93.61±11.44)比(121.01±10.23),P均<0.05]、6周时[(85.23±12.73)、(87.73±13.07)、(76.96±12.71)比(114.18±10.29),P均<0.05]、中药组和中西药联合组8周时[(79.91±13.19)、(76.17±21.18)比(94.74±19.93),P均<0.05]跟腱组织Smad5AOD均降低。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。
Objective To investigate the potential mechanism of Kunxian capsule combined with indomethacin in the treatment of a rat model of traumatic myositis ossificans.Methods A total of 96 SD rats were used to establish a model of myositis ossificans by severing the Achilles tendon and damaging the ankle joint.According to the random number table method,the rats were divided into four groups:control,traditional Chinese medicine(TCM),Western medicine,and combined TCM and Western medicine groups,with 24 rats in each group.On the day of surgery,the control,TCM,Western medicine,and combined TCM and Western medicine groups were respectively given 0.2 g/50 mL placebo,0.3 g/50 mL Kunxian capsule powder solution,0.1 g/50 mL indomethacin powder solution,0.3 g/50 mL Kunxian capsule powder mixed with 0.1 g/50 mL indomethacin powder solution by gavage,once a day,2 mL each time,for 2 to 8 weeks.At 2,4,6,and 8 weeks post-surgery,six rats from each group were euthanized by cervical dislocations.X-ray imaging was performed on the ankle joints to measure the swelling thickness.Serum levels of transforming growth factor-β(TGF-β)were detected using enzyme-linked immunosorbent assay.The protein expressions of bone morphogenetic protein 4(BMP4)and Smad5 in Achilles tendon tissue were detected using immunohistochemistry.Results In terms of swelling thickness,compared to the control group at the same time point,the combined TCM and Western medicine group showed a significant decrease at 4 weeks[(10.77±0.88)vs.(13.78±0.91)mm;P<0.05];the TCM,Western medicine,and combined TCM and Western medicine groups showed significant decreases at 6 week[(11.82±1.08),(11.34±0.88),and(8.97±0.53)vs.(12.97±0.51)mm;all P<0.05];and the TCM,Western medicine,and combined TCM and Western medicine groups showed significant decreases at 8 weeks[(9.39±0.90),(9.47±0.45),and(8.47±0.38)vs.(12.25±0.89)mm,all P<0.05].In terms of serum TGF-βlevels,compared to the control group at the same time point,the Western medicine and combined TCM and Western medicine groups showed significant decreases at 2 weeks[(386.00±29.46),(376.50±27.96)vs.(484.50±59.49)ng/L;both P<0.05];the TCM,Western medicine,and combined TCM and Western medicine groups showed significant decreases at 4 weeks[(374.33±31.48),(331.83±47.11),and(337.66±46.86)vs.(453.16±53.11)ng/L;all P<0.05];the TCM group,Western medicine,and combined TCM and Western medicine groups showed significant decreases at 6 weeks[(346.66±47.84),(304.50±26.25),and(277.33±44.79)vs.(417.00±32.58)ng/L;all P<0.05];and the TCM,Western medicine,and combined TCM and Western medicine groups showed significant decreases at 8 weeks[(307.16±24.46),(283.33±34.85),and(169.50±31.64)vs.(384.83±42.22)ng/L;all P<0.05].In terms of average optical density(AOD)of BMP4 in Achilles tendon tissue,compared to the control group at the same time point,the TCM,and combined TCM and Western medicine groups showed significant decreases at 4 weeks[(148.18±15.88),(146.01±14.33)vs.(189.41±14.40);both P<0.05];the TCM,Western medicine,and combined TCM and Western medicine groups showed significant decreases at 6 weeks[(137.00±8.95),(123.15±22.93),(112.53±35.93)vs.(172.98±7.56);all P<0.05];the TCM,Western medicine,and combined TCM and Western medicine groups showed significant decreases at 8 weeks[(128.21±12.85),(117.58±21.34),(96.04±21.33)vs.(157.95±10.41);all P<0.05].In terms of AOD of Smad5 in Achilles tendon tissue,compared to the control group at the same time point,the combined TCM and Western medicine group showed a significant decrease at 2 weeks[(101.98±9.46)vs.(130.94±9.54);P<0.05];the TCM,Western medicine,and combined TCM and Western medicine groups showed significant decreases at 4 weeks[(92.65±12.96),(96.97±12.85),(93.61±11.44)vs.(121.01±10.23);all P<0.05];the TCM group,Western medicine,and combined TCM and Western medicine groups showed significant decreases at 6 weeks[(85.23±12.73),(87.73±13.07),(76.96±12.71)vs.(114.18±10.29);all P<0.05];the TCM and combined TCM and Western medicine groups showed significant decreases at 8 weeks[(79.91±13.19),(76.17±21.18)vs.(94.74±19.93);both P<0.05].Conclusion Kunxian capsules combined with indomethacin have a synergistic inhibitory effect on traumatic myositis ossificans,and the mechanism may be related to the regulation of the BMP4/Smad5 signaling pathway.
作者
吴静泽
王晔恺
陈位
金丹雯
WU Jingze;WANG Yekai;CHEN Wei;JINDanwen(Departments of Radiology,Zhoushan Hospital,Zhoushan,Zhejiang 316021,China;Departments of Clinical Laboratory,Zhoushan Hospital,Zhoushan,Zhejiang 316021,China;Departments of Orthopedics,Zhoushan Hospital,Zhoushan,Zhejiang 316021,China;Departments of Pathology,Zhoushan Hospital,Zhoushan,Zhejiang 316021,China)
出处
《浙江中西医结合杂志》
2024年第8期703-707,共5页
Zhejiang Journal of Integrated Traditional Chinese and Western Medicine
基金
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)。
