α-SMA在骨科挛缩性疾病中的表达及其意义
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摘要
一、背景
臀肌挛缩症、先天性肌性斜颈、掌腱膜挛缩症是骨科常见软组织挛缩性疾病。均表现为病变组织挛缩而出现畸形，甚至影响功能。臀肌挛缩症多认为同局部肌肉注射有关；先天性肌性斜颈则由多种原因所致；目前对掌腱膜挛缩症的发病机制研究较多，认为肌纤维母细胞是掌腱膜挛缩症发病的重要病理因素。该细胞既有能够引起收缩的微丝样结构，又能合成分泌大量胶原及细胞外基质成分，使掌腱膜纤维化挛缩。而对臀肌挛缩症、先天性肌性斜颈发病的分子机制的研究报道较少。在臀肌挛缩症和先天性肌性斜颈中是否存在有相同的病理机制尚不清楚。因此，研究肌纤维母细胞的标记蛋白α-SMA在这些疾病中的表达、分布情况，有助于了解病理变化过程、病理机制，并指导临床治疗，以免延误病情，影响预后。
二、目的
我们应用免疫组化辅以计算机图象定量分析技术，观察和检测α-SMA在臀肌挛缩症、先天性肌性斜颈和掌腱膜挛缩症中表达和分布，以明确臀肌挛缩症和先天性肌性斜颈中是否出现肌纤维母细胞，了解病理变化过程，病理机制，为临床诊断治疗提供依据。
浙汪大学硕士学位论文
三、材料和方法
收集我院自1994年10月～2002年10月8年间收治的骨科挛缩性疾病标
本，臀肌挛缩症患者标本26例，男12例，女14例，年龄5和 岁，平均年
龄10二3岁；先天性肌性斜颈标本月例，男四例，女10例，年龄2～15岁，
平均 6．44岁；掌健膜挛缩症标本 20例，男 18例，女 2例，年龄 48～75岁，
平均 61．24岁。所有标本经病理学检查确诊。正常臀肌及筋膜 12例，年龄
6～22岁，平均11．27岁；肌肉瞻性部分标本10例，年龄6～14岁，平均8．72
岁，作为先天性肌性斜颈正常对照；正常掌位膜标本9例，3例外伤性截肢
而手腕部未损伤的手掌、2例腕部手术延伸切口的手掌、4例新鲜尸体的手
掌、作为掌健膜挛缩性正常对照。
四、统计方法
应用 SPSS10．0 for widows专业版统计软件。数据用均数士标准差（X士S）
表示，病变组与正常对照组间进行t检验，用（S——N——K）法进行三组单因
素方差分析，若差异有显著性意义，再进行各组间两两比较…检验人 以P
＜ 0刀5作为差异有显著意义。
五、结果
a－SMA在各对照组未见明显表达，而在三组病变组织中有不同程度阳
性反应，与对照组间有非常显著差异呼＜队001）。用 u一＊一K）法三组挛
缩性组织间单因素方差分析示总体有差异，但两两之间无差异一检验人臀
肌挛缩症组织，7岁以上年龄组a－SMA表达明显减弱，与7岁以下组比较差
异有显著性意义k＝2．364 p－0刀28人 先天性肌性斜颈6岁以上组表达明显
减弱，与6岁以下组（包括6岁）比较差异有非常显著意义h一5．1刀，P＜0刀01）。
－3一
浙江大学硕士学位论文
I＼、二口任匕
1、肌纤维母细胞是臀肌挛缩症，先天性肌性斜颈，掌健膜挛缩症三种疾
病共同的病理基础。
2、本研究发现a－SMA在6岁以上组先天性肌性斜颈组织中表达明显减
弱，而在臀肌挛缩症患者标本7岁以上组表达明显减弱；且臀肌挛缩症残余
期患者明显比其他二组患者多。这些给临床诊断治疗提供新的思考。
1.Background:
The gluteal muscle contracture,congenital muscular torticollis and Dupuytren's are common soft tissue contracture dieases in bone and joint surgery.The clinical manifestations of these dieases are contracture of pathologic tissue,finally result in deformity,even influence their function.Gluteal muscle contracture is related with local gluteal muscle injection.Congenital muscular torticollis result in many cases.Nowadays,there are many reports about the pathogenesis of Dupuytren's contracture. The myofibroblast is believed to be one of the important factors in Dupuytren's contracture. The cell has microfilament stracture with contractility,and can synthesize and secrete collagen and extracellular matrix(ECM), it make palma aponeurosis produce fibrosis contracture. But the pathogenesis of molecule study on gluteal muscule contracture and congenital muscular torticollis are rarely reported. If exist the same pathological pathogenesis in the gluteal muscule contracture and congenital muscular torticollis is not
clear.The alpha-smooth muscle actin is reliable marker for myofibroblastic cellular phenotye. Therefore, studying the expression and distribution of alpha-smooth muscle actin in these contracture diseases to get a better understanding of histological phase of the pathologic development, pathogenesis and guide clinical therapy in order to avoid delaying therapy and influence the prognosis.
2.Objective:
By using immunohistochemical approach combined with computer figure analysis technique we study the expression and distribution of alpha-smooth muscle actin in gluteal muscle contracture ,congenital muscular torticollis and Dupuytren's contracture .It is determine the gluteal muscle contracture and congenital muscular torticollis whether has myofibroblast and to get a better understanding of the pathologic development of the above 3 diseases,pathogenesis, and gives fresh thought to practical diagnosis and cure of these diseases.
3. Material and Method:
Samples on bony contracture cases since Oct. 1994 till Oct.2002 in our medical college were gathered. Among them, 26 were gluteal muscle contractures including 12 male cases and 14 female ones aged 5 to 17 and 10.23 on average; 21 were congenital muscular torticollis including 11 male cases and 10 female ones aged 2 to 15 and 6.44 on average;20 were Dupuytren's contractures including 18 male cases and 2 female ones aged 48 to 75 and 61.24 on average. Also, 12normal gluteal muscle and fascia samples were gathered aged 6 to 22 and 11.27 on average as a contrast to the gluteal muscle contractures; 10 tendon samples aged 6 to 14 and 8.72 on average in contrast to congenital muscular torticollis and 9 normal palma aponeurosis samples in contrast to palmar aponeurosis contractures.
4. Statistics Method:
With the help of SPSS 10.0 for Windows software, all data were indicated with X ± S and test(t) was carried out between the diseased groups and the normal ones. Meanwhile, the method of S-N-K was applied to make analysis of differences among the 3 groups in terms of each single factor. If the difference is striking, further comparison was made between each group .If P<0.05, the difference is of striking point.
5.Result: a-SMA was not obviously expressed in each contrasting group while in the diseased groups there existed positive reactions of various degrees , which carried a striking difference from their correspondent constrasting groups(P<0.001). The 3 diseased groups as a whole showed difference (S-N-K meathod) but there wasn't between every two of them (q test). In the gluteal muscle contracture group ,the expression of a-SMA of those aged over 7 weakened obviously, whose difference from those aged less than 7 was of striking importance (t=2.364,p=0.028).In the congenital muscular torticollis group, the expression of alpha-smooth muscle acthin of those aged over 6 was also weakened obviously, whose difference from those aged no more than 6(including 6) was also of great point(t=5.173,P<0.001) Conclusions:
1. Myofibroblas is c
臀肌挛缩症、先天性肌性斜颈、掌腱膜挛缩症是骨科常见软组织挛缩性疾病。均表现为病变组织挛缩而出现畸形，甚至影响功能。臀肌挛缩症多认为同局部肌肉注射有关；先天性肌性斜颈则由多种原因所致；目前对掌腱膜挛缩症的发病机制研究较多，认为肌纤维母细胞是掌腱膜挛缩症发病的重要病理因素。该细胞既有能够引起收缩的微丝样结构，又能合成分泌大量胶原及细胞外基质成分，使掌腱膜纤维化挛缩。而对臀肌挛缩症、先天性肌性斜颈发病的分子机制的研究报道较少。在臀肌挛缩症和先天性肌性斜颈中是否存在有相同的病理机制尚不清楚。因此，研究肌纤维母细胞的标记蛋白α-SMA在这些疾病中的表达、分布情况，有助于了解病理变化过程、病理机制，并指导临床治疗，以免延误病情，影响预后。
二、目的
我们应用免疫组化辅以计算机图象定量分析技术，观察和检测α-SMA在臀肌挛缩症、先天性肌性斜颈和掌腱膜挛缩症中表达和分布，以明确臀肌挛缩症和先天性肌性斜颈中是否出现肌纤维母细胞，了解病理变化过程，病理机制，为临床诊断治疗提供依据。
浙汪大学硕士学位论文
三、材料和方法
收集我院自1994年10月～2002年10月8年间收治的骨科挛缩性疾病标
本，臀肌挛缩症患者标本26例，男12例，女14例，年龄5和 岁，平均年
龄10二3岁；先天性肌性斜颈标本月例，男四例，女10例，年龄2～15岁，
平均 6．44岁；掌健膜挛缩症标本 20例，男 18例，女 2例，年龄 48～75岁，
平均 61．24岁。所有标本经病理学检查确诊。正常臀肌及筋膜 12例，年龄
6～22岁，平均11．27岁；肌肉瞻性部分标本10例，年龄6～14岁，平均8．72
岁，作为先天性肌性斜颈正常对照；正常掌位膜标本9例，3例外伤性截肢
而手腕部未损伤的手掌、2例腕部手术延伸切口的手掌、4例新鲜尸体的手
掌、作为掌健膜挛缩性正常对照。
四、统计方法
应用 SPSS10．0 for widows专业版统计软件。数据用均数士标准差（X士S）
表示，病变组与正常对照组间进行t检验，用（S——N——K）法进行三组单因
素方差分析，若差异有显著性意义，再进行各组间两两比较…检验人 以P
＜ 0刀5作为差异有显著意义。
五、结果
a－SMA在各对照组未见明显表达，而在三组病变组织中有不同程度阳
性反应，与对照组间有非常显著差异呼＜队001）。用 u一＊一K）法三组挛
缩性组织间单因素方差分析示总体有差异，但两两之间无差异一检验人臀
肌挛缩症组织，7岁以上年龄组a－SMA表达明显减弱，与7岁以下组比较差
异有显著性意义k＝2．364 p－0刀28人 先天性肌性斜颈6岁以上组表达明显
减弱，与6岁以下组（包括6岁）比较差异有非常显著意义h一5．1刀，P＜0刀01）。
－3一
浙江大学硕士学位论文
I＼、二口任匕
1、肌纤维母细胞是臀肌挛缩症，先天性肌性斜颈，掌健膜挛缩症三种疾
病共同的病理基础。
2、本研究发现a－SMA在6岁以上组先天性肌性斜颈组织中表达明显减
弱，而在臀肌挛缩症患者标本7岁以上组表达明显减弱；且臀肌挛缩症残余
期患者明显比其他二组患者多。这些给临床诊断治疗提供新的思考。
1.Background:
The gluteal muscle contracture,congenital muscular torticollis and Dupuytren's are common soft tissue contracture dieases in bone and joint surgery.The clinical manifestations of these dieases are contracture of pathologic tissue,finally result in deformity,even influence their function.Gluteal muscle contracture is related with local gluteal muscle injection.Congenital muscular torticollis result in many cases.Nowadays,there are many reports about the pathogenesis of Dupuytren's contracture. The myofibroblast is believed to be one of the important factors in Dupuytren's contracture. The cell has microfilament stracture with contractility,and can synthesize and secrete collagen and extracellular matrix(ECM), it make palma aponeurosis produce fibrosis contracture. But the pathogenesis of molecule study on gluteal muscule contracture and congenital muscular torticollis are rarely reported. If exist the same pathological pathogenesis in the gluteal muscule contracture and congenital muscular torticollis is not
clear.The alpha-smooth muscle actin is reliable marker for myofibroblastic cellular phenotye. Therefore, studying the expression and distribution of alpha-smooth muscle actin in these contracture diseases to get a better understanding of histological phase of the pathologic development, pathogenesis and guide clinical therapy in order to avoid delaying therapy and influence the prognosis.
2.Objective:
By using immunohistochemical approach combined with computer figure analysis technique we study the expression and distribution of alpha-smooth muscle actin in gluteal muscle contracture ,congenital muscular torticollis and Dupuytren's contracture .It is determine the gluteal muscle contracture and congenital muscular torticollis whether has myofibroblast and to get a better understanding of the pathologic development of the above 3 diseases,pathogenesis, and gives fresh thought to practical diagnosis and cure of these diseases.
3. Material and Method:
Samples on bony contracture cases since Oct. 1994 till Oct.2002 in our medical college were gathered. Among them, 26 were gluteal muscle contractures including 12 male cases and 14 female ones aged 5 to 17 and 10.23 on average; 21 were congenital muscular torticollis including 11 male cases and 10 female ones aged 2 to 15 and 6.44 on average;20 were Dupuytren's contractures including 18 male cases and 2 female ones aged 48 to 75 and 61.24 on average. Also, 12normal gluteal muscle and fascia samples were gathered aged 6 to 22 and 11.27 on average as a contrast to the gluteal muscle contractures; 10 tendon samples aged 6 to 14 and 8.72 on average in contrast to congenital muscular torticollis and 9 normal palma aponeurosis samples in contrast to palmar aponeurosis contractures.
4. Statistics Method:
With the help of SPSS 10.0 for Windows software, all data were indicated with X ± S and test(t) was carried out between the diseased groups and the normal ones. Meanwhile, the method of S-N-K was applied to make analysis of differences among the 3 groups in terms of each single factor. If the difference is striking, further comparison was made between each group .If P<0.05, the difference is of striking point.
5.Result: a-SMA was not obviously expressed in each contrasting group while in the diseased groups there existed positive reactions of various degrees , which carried a striking difference from their correspondent constrasting groups(P<0.001). The 3 diseased groups as a whole showed difference (S-N-K meathod) but there wasn't between every two of them (q test). In the gluteal muscle contracture group ,the expression of a-SMA of those aged over 7 weakened obviously, whose difference from those aged less than 7 was of striking importance (t=2.364,p=0.028).In the congenital muscular torticollis group, the expression of alpha-smooth muscle acthin of those aged over 6 was also weakened obviously, whose difference from those aged no more than 6(including 6) was also of great point(t=5.173,P<0.001) Conclusions:
1. Myofibroblas is c
引文
1.Majno G et al: Science 1971 173: 39967: 548-50.
2.Gabbiani Get al: J Exp Med 1972 135(4):719-34.
3.Magro,G: Fraggetta, -F: Myofibroblast and extracellular matrix glycoproteinsin palmar fibromatosis. Gen-Diagn-Pathol. 1997 Feb: 14(3～4): 185-90.
4.Schurch W, Seemayer TA, Gabbiani G, myofibroblast. In: Sternberg SS, ed. Histology for Pathologist. New York:Raven Press, 1992,109～144.
5.Ronnov-Jessen L, Petersen ow. A founction for filamentous α-Smooth muscle actin: retardation of motility in fibroblasts. J Cell Biol, 1996,134:67.
6.Luck JV. Dupuytren's contracture; a new concept of the pathogenesis appelated with surgical management. J Bone Joint Surg (Am), 1959, 41:635:664.
7.Baird KS, Crossan JF, Ralston SH, Abnormdal growth factor and cytokine expression in Dupuyten's contracture. J clin Pathol, 1993,46:425-428.
8.Robert M, MeFarlane. M.D. The current status of Dupuytren's disease. J Hand Surg, 1953, 9:703.
9.Wilutzky,-B; Berndt, -A; Programmed cell death in nodulai palmar fibromatosis (morbus Dupuytren). Histol-Histopthol. 1998-Jan: 13(1):67-72.
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ed. Histology for Pathologist, Raven Press. New York, 1992,109-144.
40．侯明钟，袁启智，等， 国人掌腱膜挛缩症的诱因及显微外科治疗[J]，中华显微外科杂志，1999，21(1)：58．
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43．潘志军，黄宗坚等，掌腱膜挛缩症的临床及病理基础，中国矫形外科杂志，2000年9月第7卷第9期。
2.Gabbiani Get al: J Exp Med 1972 135(4):719-34.
3.Magro,G: Fraggetta, -F: Myofibroblast and extracellular matrix glycoproteinsin palmar fibromatosis. Gen-Diagn-Pathol. 1997 Feb: 14(3～4): 185-90.
4.Schurch W, Seemayer TA, Gabbiani G, myofibroblast. In: Sternberg SS, ed. Histology for Pathologist. New York:Raven Press, 1992,109～144.
5.Ronnov-Jessen L, Petersen ow. A founction for filamentous α-Smooth muscle actin: retardation of motility in fibroblasts. J Cell Biol, 1996,134:67.
6.Luck JV. Dupuytren's contracture; a new concept of the pathogenesis appelated with surgical management. J Bone Joint Surg (Am), 1959, 41:635:664.
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