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[不同时长静态渐进性拉伸对大鼠创伤性膝关节挛缩的治疗作用及机制]

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

作者信息

Wang Lu, Zhang Li-Ning, He Jia-le, Zuo Xiu-Qin, Xie Hui-Min, Jia Zi-Shan

机构信息

Center of Rehabilitation Medicine, the First Medical Center, Chinese PLA General Hospital, Beijing 100853, China.

Chinese PLA Medical School, Beijing 100853, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2020 Mar;51(2):185-192. doi: 10.12182/20200360204.

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 ( =50), control group (CON, no surgery, no treatment, =10) and trauma without immobilization group (TRA, no treatment, =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, =10), treatment for 30 minutes group (S30 min, =10), treatment for 40 minutes group (S40 min, =10), untreatment group (UNT, no SPS, =10) and modeling group (MOD, =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 ( >0.05), and was the best among all the surgical modeling groups after 16 d of treatment ( <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 ( <0.01). ② Gait analysis: the stands in the S30min group improved best on the day 8 and day 16 ( <0.05) , and better than that on day 0 ( <0.05). The stride length of the S30 min group progressed similar to that of the S40 min group on the day 8 ( >0.05), and there was no difference among three groups on the day 16 ( >0.05). The stride length of the S30 min group appeared to recover more quickly on the day 8 ( <0.05), and those of S20 min and UNT groups recovered significantly on the day 16 ( <0.05). In addition, the swings in the S30 min group improved best ( <0.05), and it appeared to recover better on the day 16 ( <0.05). There was no statistical difference in terms of the swing speed among the four surgical modeling groups on the day 8 ( >0.05). The swing speed of the S30min group increased most than those of the other three groups ( <0.05), and it was much better on the day 8 and day 16 comparing with that on the day 0 ( <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 ( <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.

摘要

目的

探讨不同时长的静态渐进性拉伸(SPS)对大鼠创伤性膝关节挛缩的影响及其机制。

方法

70只雄性Wistar大鼠随机分为三组,包括手术建模组(n = 50)、对照组(CON,未手术、未治疗,n = 10)和未固定创伤组(TRA,未治疗,n = 10)。建立膝关节挛缩模型,将50只手术建模大鼠随机分为五组,包括静态渐进性拉伸治疗20分钟组(S20 min,n = 10)、治疗30分钟组(S30 min,n = 10)、治疗40分钟组(S40 min,n = 10)、未治疗组(UNT,未进行SPS,n = 10)和建模组(MOD,n = 10,固定后安乐死用于组织学染色和蛋白质印迹法)。S20 min、S30 min和S40 min组的大鼠麻醉后进行SPS。每隔一天进行一次治疗。直至第16天的最后一次治疗,共进行8次治疗。在干预的第0、8和16天,测量并比较关节活动范围(ROM)和步态分析。在ROM测量和步态分析后,于第16天对大鼠实施安乐死,样本采用苏木精-伊红(HE)和马松(Masson)方法染色。观察病理组织学变化。采用蛋白质印迹法检测转化生长因子-β1(TGF-β1)和白细胞介素-6(IL-6)的表达。

结果

① ROM:治疗8天后,S30 min组的ROM恢复情况与S20 min和S40 min组相似(P > 0.05),且在治疗16天后是所有手术建模组中恢复最佳的(P < 0.05)。与第0天相比,S20 min、S30 min和S40 min组在第8天和第16天的ROM显著改善(P < 0.01)。② 步态分析:S30min组在第8天和第16天的站立情况改善最佳(P < 0.05),且优于第0天(P < 0.05)。S30 min组在第8天的步幅进展与S40 min组相似(P > 0.05),在第16天三组之间无差异(P > 0.05)。S30 min组在第8天的步幅恢复似乎更快(P < 0.05),S20 min组和UNT组在第16天步幅显著恢复(P < 0.05)。此外,S30 min组的摆动情况改善最佳(P < 0.05),且在第16天似乎恢复更好(P < 0.05)。在第8天,四个手术建模组的摆动速度无统计学差异(P > 0.05)。S30min组的摆动速度比其他三组增加最多(P < 0.05),且在第8天和第16天与第0天相比有显著改善(P < 0.05)。③ HE和Masson染色:在第16天,与其他组相比,S30min组的纤维化和炎症明显受到抑制。④ 蛋白质印迹法:在第16天,TGF-β1和IL-6的蛋白表达水平显著低于其他干预组,包括S20 min、S40 min和UNT组(P < 0.05)。

结论

30分钟的静态渐进性拉伸治疗可显著改善大鼠创伤性膝关节挛缩。其机制可能是SPS降低了TGF-β1和IL-6的表达,减少了关节囊的粘连和炎症。因此,通过重建关节囊结构并抑制纤维化改变,缓解了疼痛并增加了关节活动度。

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