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不同时长的静态进展性牵伸对大鼠创伤性膝关节挛缩的治疗效果及其机制研究

The Therapeutic Effect and Mechanism of Static Progressive Stretching in Different Durations on Traumatic Knee Contracture in Rats

  • 摘要:
      目的  探讨不同时长的静态进展性牵伸治疗大鼠创伤性膝关节挛缩的效果及机制。
      方法  选取雄性Wistar大鼠70只,随机分为手术造模组50只,空白对照组(不造模,不治疗)10只和单纯创伤未制动组(造成创伤不加克氏针固定,不治疗)10只。手术造模组50只建立膝关节挛缩模型,再随机分为静态进展性牵伸治疗20 min组(S20 min),治疗30 min组(S30 min),治疗40 min组(S40 min),非牵伸组(造模后只麻醉,不牵伸)以及模型对照组(造模后即刻处死,只用于病理染色和蛋白检测)共5组,每组各10只。牵伸治疗周期为隔天一次,牵伸治疗8次,共16 d。于治疗0 d、8 d、16 d测量各组关节活动度(ROM)、步态分析,于治疗16 d后处死大鼠,取膝关节后囊组织进行HE及Masson染色,比较组织病理变化,Western blot检测转化生长因子(TGF)-β1与白介素(IL)-6的表达。
      结果  ① ROM:S30 min组治疗8 d时,ROM恢复与S20 min、S40 min组相当(P>0.05),治疗16 d时则恢复最佳(P<0.05);治疗8 d、16 d时,S20 min、S30 min、S40 min组ROM均较0 d改善(P<0.01)。② 步态分析:S30 min组治疗8 d和16 d时支撑相改善最佳(P<0.05),且均较0 d改善(P<0.05)。S30 min组治疗8 d时和S40 min组步幅改善相似(P>0.05),治疗16 d时和S20 min、S40 min步幅改善相似(P>0.05);S30 min组治疗8 d时步幅较0 d有明显改善(P<0.05),16 d时改善不明显。治疗8 d和16 d时,S30 min组摆动相恢复最佳(P<0.05),治疗8 d较0 d改善不明显,治疗16 d时摆动相较0 d有改善(P<0.05)。S30 min组治疗8 d时摆动速度与S20 min组、S40 min组、非牵伸组差异无统计学意义(P>0.05),治疗16 d摆动速度改善优于其他3个治疗组(P<0.05);治疗8 d、16 d时S30 min组摆动速度较0 d有改善(P<0.05)。③ HE染色及Masson染色:治疗16 d后,S30 min组关节囊纤维组织增生及炎症改善最明显。④ Western blot检测:治疗16 d后,S30 min组TGF-β1和IL-6的蛋白表达低于S20 min组、S40 min组、非牵伸组,差异有统计学意义(P<0.05)。
      结论  静态进展性牵伸治疗30 min对大鼠创伤性膝关节挛缩改善最明显,其机制为降低TGF-β1与IL-6的表达,减轻关节粘连和炎症反应,重塑关节囊结构,抑制关节囊纤维化,缓解疼痛,提高关节活动功能。

     

    Abstract:
      Objective  To investigate the effect and mechanism of static progressive stretching (SPS) in different durations on traumatic knee contracture in rats.
      Methods  Seventy male Wistar rats were randomly divided into three groups, including surgical modeling group (n=50), control group (CON, no surgery, no treatment, n=10) and trauma without immobilization group (TRA, no treatment, n=10). The knee contracture model was established, and 50 surgical modeling rats were randomly divided into five groups including static progressive stretching treatment for 20 minutes group (S20 min, n=10), treatment for 30 minutes group (S30 min, n=10), treatment for 40 minutes group (S40 min, n=10), untreatment group (UNT, no SPS, n=10) and modeling group (MOD, n=10, euthanized after immobilization for histological staining and Western blot). Individuals in the S20 min, S30 min, and S40 min groups were anesthetized and submitted to SPS. One treatment session took place every other day. A total of 8 sessions were given till the final treatment session on the day 16. On the day 0, 8, and 16 of intervention, the range of joint motion (ROM) and gait analysis were measured and compared. After the ROM measurements and gait analysis, the rats were euthanized on the day 16 and the samples were stained with HE and Masson methods. The changes of pathological organization were observed. Western blot was used to detect the expressions of transforming growth factor-β1 (TGF-β1) and interleukin-6 (IL-6).
      Results  ① ROM: the ROM of S30 min group recovered similar to that of the S20 min and S40 min groups after 8 days of treatment (P>0.05), and was the best among all the surgical modeling groups after 16 d of treatment (P<0.05). The ROM of S20 min, S30 min and S40 min groups significantly improved on the day 8 and day 16 comparing with that on day 0 (P<0.01). ② Gait analysis: the stands in the S30min group improved best on the day 8 and day 16 (P<0.05) , and better than that on day 0 (P<0.05). The stride length of the S30 min group progressed similar to that of the S40 min group on the day 8 (P>0.05), and there was no difference among three groups on the day 16 (P>0.05). The stride length of the S30 min group appeared to recover more quickly on the day 8 (P<0.05), and those of S20 min and UNT groups recovered significantly on the day 16 (P<0.05). In addition, the swings in the S30 min group improved best (P<0.05), and it appeared to recover better on the day 16 (P<0.05). There was no statistical difference in terms of the swing speed among the four surgical modeling groups on the day 8 (P>0.05). The swing speed of the S30min group increased most than those of the other three groups (P<0.05), and it was much better on the day 8 and day 16 comparing with that on the day 0 (P<0.05 ). ③ HE and Masson staining: the fibrosis and inflammation of the S30min group were significantly suppressed comparing to the other groups on the day 16. ④ Western blot: The protein expression levels of TGF-β1 and IL-6 were significantly lower than those in the other intervention groups including the S20 min, S40 min and UNT groups on the day 16 (P<0.05).
      Conclusion  Static progressive stretching treatment for 30 min could significantly improve the traumatic knee contracture in rats. The mechanism may be that the SPS decreased the expressions of TGF-β1 and IL-6, reduced the adhesion and inflammation of joint capsule. Therefore it relieved the pain and increased the joint mobility by reconstructing the structure of the capsule and suppressing the fibrotic changes.

     

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