Differentially expressed proteins on postoperative 3 days healing in rabbit Achilles tendon rupture model after early kinesitherapy

Objectives: Surgical repair of Achilles tendon (AT) rupture should immediately be followed by active tendon mobilization. The optimal time as to when the mobilization should begin is important yet controversial.Early kinesitherapy leads to reduced rehabilitation period.However, an insight into the detailed mechanism of this process has not been gained. Proteomic technique can be used to separate and purify the proteins by differential expression profile which is related to the function of different proteins, but research in the area ofproteomic analysis of AT 3 days after repair has not been studied so far.Methods: Forty-seven New Zealand white rabbits were randomized into 3 groups. Group A (immobilization group,n=16) received postoperative cast immobilization; Group B (early motion group, n= 16) received early active motion treatments immediately following the repair of AT rupture from tenotomy. Another 15 rabbits served as control group (Group C). The AT samples were prepared 3 days following the microsurgery. The proteins were separated employing twodimensional polyacrylamide gel electrophoresis (2D-PAGE).PDQuest software version 8.0 was used to identify differentially expressed proteins, followed by peptide mass fingerprint (PMF) and tandem mass spectrum analysis, using the National Center for Biotechnology Information (NCBI) protein database retrieval and then for bioinformatics analysis.Results: Amean of 446.33,436.33 and 462.67 protein spots on Achilles tendon samples of 13 rabbits in Group A,14 rabbits in Group B and 13 rabbits in Group C were successfully detected in the 2D-PAGE. There were 40, 36 and 79unique proteins in Groups A, B and C respectively. Some differentially expressed proteins were enzyme with the gel,matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). We successfully identified 9 and 11 different proteins in Groups A and B,such as GAPDH, phosphoglycerate kinase 1, pro-alpha-1type 1 collagen, peroxiredoxin 1, alpha-1-antiproteinase E a-1and MAD2L1 binding protein, etc. And some with the molecular chaperone, oxidative stress, energy metabolism,signal transduction, coupled with the tendon cell expression and protein synthesis, proliferate, differentiate and are closely related to the AT healing. The GAPDH protein was further validated through Western blotting. It was indicated that some differentially expressed proteins were involved in various metabolism pathways and may play an important role in initial healing of AT rupture.Conclusion: Differentially expressed proteins in rabbit healing AT model may contribute to 3 days healing of AT rupture through a new mechanobiological mechanism due to the application of postoperative early kinesitherapy.     

Kazakh Therapy on Differential Protein Expression of Achilles Tendon Healing in a 7-Day Postoperative Rabbit Model

Objective: To compare the effect of cast immobilization with that of early Kiymil arkili emdew (Kazakh exercise therapy) on the post-operative healing of Achilles tendon rupture in rabbits, and to observe the influence of early Kiymil arkili emdew on the differentially expressed proteins in the healing tendon. Methods: Forty-five New Zealand white rabbits were randomly divided into three groups (Arm A: control group; Arm B: postoperative immobilization group; and Arm C: postoperative early Kiymil arkili emdew group). After tenotomy, the rabbits of the two experimental groups received microsurgery to repair the ruptured tendons, and then received either cast immobilization or early Kiymil arkili emdew treatment. Achilles tendon tissue samples were collected 7 days after the surgery, and two-dimensional gel electrophoresis and MALDI-TOF-MS technique were used to analyze differentially expressed proteins in the tendon tissue of the three Arms. Results: A total of 462.67±11.59, 532.33±27.79, and 515.33±6.56 protein spots were detected by the two-dimensional polyacrylamide gels in the Achilles tendon samples of the rabbits in Arms A, B, and C, respectively. Nineteen differentially expressed protein spots were randomly selected from Arm C. Among them, 7 were unique, and 15 had five times higher abundance than those in Arm B. These included annexin A2, gelsolin isoforms and α-1 Type III collagen. It was confirmed by western blot that gelsolin isoform b, annexin A2, etc. had specific and incremental expression in Arm C. Conclusions: The self-protective instincts of humans were overlooked in the classical postoperative treatment for Achilles tendon rupture with cast immobilization. Kiymil arkili emdew induced the specific and incremental expression of proteins in the repaired Achilles tendon in the early healing stage in a rabbit model, compared with those treated with postoperative cast immobilization. These differentially expressed proteins may contribute to the healing of the Achilles tendon via a mechanobiological mechanism caused by the application of Kiymil arkili emdew.     

异丙酚对急性呼吸窘迫综合征时肾Gq/11蛋白的干预作用

目的探讨异丙酚是否通过影响Gq/11蛋白的表达对肾脏起保护作用。方法采用尾静脉注射油酸(OA)法复制急性呼吸窘迫综合征(ARDS)大鼠模型。将健康雄性Wistar大鼠随机分为对照组(C组)、OA组和异丙酚预处理组(P组),并观察血压及相关酶的变化。用蛋白质免疫印迹法检测肾脏中Gq/11蛋白含量。结果P组Gq/11蛋白含量(124.68±19.38)%较C组(100.00±0)%增加了(24.68±19.38)%(P<0.01),但明显低于OA组(149.34±20.04)%(P<0.01)。P组肾组织血管紧张素转换酶(16.52±1.37)μmol·min-·g-1、乳酸脱氢酶(1.20±0.16)kU/g活性和丙二醛(1.51±0.35)μmol/g含量也均明显低于OA组分别为(17.56±1.02)μmol·min-1·g-1,(1.41±0.16)kU/g,(1.94±0.16)μmol/g,P均<0.01。结论预先注射异丙酚能降低Gq/11蛋白在肾脏中的含量,并减轻ARDS时肾损害的程度。     

Kazakh therapy on differential protein expression of Achilles tendon healing in a 7-day postoperative rabbit model

Objective: To compare the effect of cast immobilization with that of early Kiymil arkili emdew (Kazakh exercise therapy) on the post-operative healing of Achilles tendon rupture in rabbits,and to observe the influence of early Kiymil arkili emdew on the differentially expressed proteins in the healing tendon.Methods: Forty-five New Zealand white rabbits were randomly divided into three groups (Arm A: control group;Arm B: postoperative immobilization group; and Arm C: postoperative early Kiymil arkili emdew group).After tenotomy,the rabbits of the two experimental groups received microsurgery to repair the ruptured tendons,and then received either cast immobilization or early Kiymil arkili emdew treatment.Achilles tendon tissue samples were collected 7 days after the surgery,and two-dimensional gel electrophoresis and MALDI-TOF-MS technique were used to analyze differentially expressed proteins in the tendon tissue of the three Arms.Results: A total of 462.67+11.59,532.33+27.79,and 515.33+6.56 protein spots were detected by the two-dimensional polyacrylamide gels in the Achilles tendon samples of the rabbits in Arms A,B,and C,respectively.Nineteen differentially expressed protein spots were randomly selected from Arm C.Among them,7 were unique,and 15 had five times higher abundance than those in Arm B.These included annexin A2,gelsolin isoforms and α-1 Type Ⅲ collagen.It was confirmed by western blot that gelsolin isoform b,annexin A2,etc.had specific and incremental expression in Arm C.Conclusions: The self-protective instincts of humans were overlooked in the classical postoperative treatment for Achilles tendon rupture with cast immobilization.Kiymil arkili emdew induced the specific and incremental expression of proteins in the repaired Achilles tendon in the early healing stage in a rabbit model,compared with those treated with postoperative cast immobilization.These differentially expressed proteins may contribute to the healing of the Achilles tendon via a mechanobiological mechanism caused by the application of Kiymil arkili emdew.