先天性肌性斜颈病变组织中细胞外间质的变化特点

发布时间：2018-08-21 08:11

【摘要】：目的：观察先天性肌性斜颈(Congenital Muscular Torticollis, CMT)病变组织中脂肪增生现象及分布特点,分析脂肪增生程度与纤维化和年龄的关系。方法：对手术治疗,切除的病理组织,常规石蜡包埋,并取部分新鲜标本行液氮冰冻处理保存。石蜡标本行常规HE染色,观察先天性肌性斜颈病变胸锁乳突肌基本病理改变和脂肪增生现象。Masson染色,观察胞外基质的纤维化情况。冰冻标本行苏丹Ⅲ脂肪染色,定性病变肌组织中增生的脂肪组织。结果：188例CMT患儿中,男123例,女65例,男女比约2：1；左侧75例,右侧113例,左右比约2：3；冰冻标本31例；年龄最小2月24天,最大16岁,平均年龄2.9岁。所有标本均符合CMT诊断标准及排除其它疾病。①HE染色下CMT病变胸锁乳突肌肌纤维大小不等,呈萎缩、肥大、坏死等多种改变,部分肌细胞有空泡变性和核中心化表现,间质增生纤维化明显。②Masson染色显示肌内膜、肌束膜轻重不一的纤维化增生,包绕肌纤维。随着年龄的增长,纤维化程度呈现两头高中间低的特点,0-3岁纤维化与年龄负相关(r=-0.168,P=0.049),3岁后纤维化与年龄正相关(r=0.281,P=0.001)。③苏丹Ⅲ脂肪染色可见橙红色阳性分布区域,与HE染色下脂肪分布相对应。④存在脂肪增生的标本占总标本数的69.68%(131/188),增生程度轻重不一,标本中可见少量的脂肪细胞,也可见大量脂肪增生；增生的脂肪组织多呈团块状或条索状；大多为纤维化胶原所包绕,也可见穿插于肌纤维间。脂肪增生随纤维化加重和年龄增大呈增多趋势,且三者间相互具有正相关关系(P0.05)。结论：CMT的基本病变是病变的胸锁乳突肌纤维化,纤维化程度随年龄增长呈现两头高中间低的特点。本研究在常规观察基本病变组织中,发现了脂肪增生现象。脂肪增生参与CMT的基本病理变化过程,随病变胸锁乳突肌纤维化进程而加重,并可能对肌纤维的病理改变与组织纤维化产生影响。目的：观察先天性肌性斜颈(congenital muscular torticollis, CMT)病变组织中肌纤维基底膜的变化特点,分析基底膜改变程度与对应肌纤维形态改变和间质纤维化程度的关系,推测基底膜的改变与先天性肌性斜颈病情发展与改变的内在联系。方法：通过手术治疗,切除40例先天性肌性斜颈患儿病变胸锁乳突肌(sternocleidomastoid muscle, SCM)组织及5例发育性髋关节脱位(developmental dysplasia of the hip, DDH)患儿的内收肌组织,常规石蜡包埋,切片后行HE染色、Masson染色和针对Ⅳ型胶原和层粘连蛋白的免疫组织化学染色,以发育性髋关节脱位患儿手术切除的内收肌作正常对照。观察Ⅳ型胶原和层粘连蛋白的表达分布特点,并根据Ⅳ型胶原和(或)层粘连蛋白的免疫组织化学染色所显示的肌纤维基底膜形态,分析同一标本不同病变区域的基底膜改变与对应肌纤维形态变化的关系,分析不周标本中基底膜改变与间质纤维化程度的关系。结果：HE染色下,CMT病变组织镜下观察可见明显的间质增生纤维化,残存肌纤维被增生的纤维化组织包绕并分离,肌纤维退行性变明显；但同一标本的不同病变区域纤维化程度与肌纤维残存数量不一,部分标本的病变组织可分为纤维化区、肌细胞纤维混合区和肌细胞密集区。Masson染色显示,对照组DDH内收肌,平均纤维化为8.74±1.54(%),先天性肌性斜颈病变组织中纤维化程度明显,为55.06±17.82(%),经两独立样本T检验分析,t=15.97,P0.01。在CMT中,蓝色的纤维化区域呈弥漫性增生,肌内膜、肌束膜均有轻重不一的纤维化,包绕残存肌纤维。免疫组织化学染色结果显示Ⅳ型胶原和层粘连蛋白二者的组织内表达部位基本一致,即表达在肌纤维基底膜和血管壁上,而在CMT病变组织间质表达不明显。对照组中肌纤维形态规则,大小均一,基底膜边缘光滑,薄且均匀,并紧贴与肌纤维胞膜上,不能区分。同一CMT标本不同病变区域肌纤维及其基底膜变化特点是,①纤维化区,肌纤维零星,Ⅳ型胶原和层粘连蛋白的阳性表达部位仅存在于视野中的血管壁上,而其他区域的染色结果为阴性；仅有零星肌纤维的基底膜上,未见表达,完全缺失。②肌细胞纤维混合区,残存肌纤维被增生的纤维化组织分割,呈孤岛状态,肌细胞形态不规整,常见肥大、萎缩的表现,部分肌纤维有明显的核中心化、空泡变性等退行性变,并且这种变化在基底膜缺损严重的肌细胞表现更为突出。相应的肌纤维基底膜在部分肌纤维周围有极少量的阳性表达,缺损明显,且不连续：一些部位基底膜显像弥漫性减弱,表达不连续,有局部缺失。③肌细胞密集区,肌纤维数量较丰富,基底膜表达较好,连续但不平整,不均一。不同CMT标本间肌纤维基底膜缺损程度与间质纤维化程度对比,经单因素方差分析结果提示基底膜缺损越重的标本中,间质纤维化程度越高(P0.05)。结论：间质增生纤维化是先天性肌性斜颈的基本病理改变,在纤维化的基础上病变组织中肌纤维基底膜表现出轻重不一的缺损特点。同一标本的不同病变区域基底膜及其对应肌纤维的形态改变存在差异,肌细胞密集区,Ⅳ型胶原和层粘连蛋白染色出较为正常的基底膜。基底膜染色缺损程度与间质纤维化程度相关,即纤维化越重,缺损越明显。推测肌纤维基底膜的变化特点,反应了生后CMT-SCM的病理演变过程。在肌细胞密集区较为正常的表达,支持肌细胞较为正常成肌；在混合区和纤维化区其完整性破坏的表现,可能是CMT-SCM内肌纤维的胞体不稳定性的表现,肌纤维退行性变不断加重,有效肌纤维数量减少,纤维化加重。并且缺失的基底膜对SCM内肌卫星细胞和(或)成肌细胞的保护作用降低,致使生后SCM的肌形成和再生能力降低,SCM新生肌纤维得不到补充,间质纤维程度继续加重,可导致CMT病情继续发展；反之,如果基底膜完整性相对正常,是肌细胞相对正常的表现,可能是CMT-SCM内肌纤维生命力维持的原因之一,也可能为CMT的临床自愈的表现。
[Abstract]:Objective: To observe the fatty hyperplasia and its distribution in congenital muscular torticollis (CMT) and analyze the relationship between the degree of fatty hyperplasia and fibrosis and age.
Methods: Surgical treatment, excision of pathological tissue, routine paraffin embedding, and some fresh specimens were frozen and preserved in liquid nitrogen. Paraffin specimens were stained with routine HE to observe the basic pathological changes and fat hyperplasia of sternocleidomastoid muscle in congenital muscular torticollis. Masson staining was used to observe the fibrosis of extracellular matrix. Sultan fatty acid staining was used to identify adipose tissue in pathological muscle tissue.
Results: Among 188 children with CMT, 123 were males and 65 were females, the ratio of male to female was about 2:1; 75 were left, 113 were right, the ratio of left to right was about 2:3; 31 were frozen specimens; the minimum age was 24 days, the maximum age was 16 years, and the average age was 2.9 years. (2) Masson staining showed the fibrosis and hyperplasia of endomyocardium and myofibrillar membrane with different degrees, wrapping the myofibrils. With the increase of age, the degree of fibrosis showed the characteristics of both ends, high school and low, fibrosis and fibrosis in 0-3 years old. Age was negatively correlated (r = - 0.168, P = 0.049), and fibrosis was positively correlated with age after 3 years old (r = 0.281, P = 0.001). 3 Sudan III fat staining showed an orange-red positive distribution, which corresponded to the distribution of fat under HE staining. Cells, but also a large number of adipose hyperplasia; hyperplastic adipose tissue is mostly lumpy or banded; most of them are wrapped by fibrotic collagen, but also can be seen interpenetrating between muscle fibers.
Conclusion: The basic pathological changes of CMT are fibrosis of the sternocleidomastoid muscle, and the degree of fibrosis is lower in both ends of the sternocleidomastoid muscle with age. It may also affect the pathological changes of muscle fibers and tissue fibrosis.
Objective: To observe the changes of myofibrillar basement membrane (MBM) in congenital muscular torticollis (CMT) lesions and to analyze the relationship between the changes of MBM and the morphological changes of corresponding myofibrils and the degree of interstitial fibrosis.
Methods: Forty children with congenital muscular torticollis were treated with surgical excision of sternocleidomastoid muscle (SCM) and five children with developmental dysplasia of the hip (DDH). The adductor muscles were embedded in paraffin and stained with HE, Masson and for type IV collagen and DDH. Immunohistochemical staining of laminin was performed in adductor muscles of children with developmental dislocation of the hip. The expression and distribution of type IV collagen and laminin were observed. The same marker was analyzed according to the morphology of basement membrane of myofibrils revealed by immunohistochemical staining of type IV collagen and/or laminin. The relationship between the changes of basement membrane and the morphological changes of myofibrils in different lesion areas was analyzed.
Results: Under HE staining, there were obvious interstitial hyperplasia and fibrosis in CMT lesions, and the residual muscle fibers were surrounded and separated by hyperplastic fibrosis tissues, and the degeneration of muscle fibers was obvious. Masson staining showed that the average fibrosis of adductor muscles in the control group was 8.74 (%) and that of congenital torticollis was 55.06 (%). Immunohistochemical staining showed that type IV collagen and laminin were expressed in the basement membrane and vascular wall of myofibrils, but not in the interstitial tissue of CMT lesions. The changes of myofibrils and their basement membranes in different lesions of the same CMT specimen were characterized by: (1) the positive expression of myofibrils, sporadic myofibrils, type IV collagen and laminin only existed in the vascular wall of the visual field, but not in the same CMT specimen. The staining results of other areas were negative; only sporadic myofibrils were found on the basement membrane, but no expression was found and completely absent. The corresponding basement membrane of myofibrils had a few positive expression around some myofibrils, and the defect was obvious and discontinuous. The basement membrane imaging of some parts of the myofibrils was diffuse weakening, expression was discontinuous, and there were local deletions. The number of fibers was abundant, the expression of basement membrane was good, continuous but uneven, and uneven. The degree of fibrosis and defect of myofibrillar basement membrane in different CMT specimens were compared. The results of one-way ANOVA analysis showed that the more serious the defect of basement membrane, the higher the degree of interstitial fibrosis (P 0.05).
Conclusion: Interstitial hyperplasia and fibrosis are the basic pathological changes of congenital muscular torticollis. The basement membrane of myofibrillar basement membrane in the pathological tissues of congenital muscular torticollis shows different characteristics on the basis of fibrosis. The degree of defect of basement membrane staining is related to the degree of interstitial fibrosis, that is, the more fibrosis, the more obvious the defect. It is speculated that the changes of basement membrane of muscle fibers reflect the pathological evolution of postnatal CMT-SCM. In the mixed and fibrotic regions, the integrity of the myofibrils may be destroyed by the instability of the cytoplasm, the degeneration of myofibrils, the decrease of the number of effective myofibrils and the aggravation of fibrosis, and the decrease of the protective effect of the missing basement membrane on the satellite cells and/or myoblasts in the SCM, resulting in the decrease of postnatal SC. If the integrity of basement membrane is relatively normal, it is a relatively normal expression of myocytes, which may be one of the reasons for maintaining the viability of myofibers in CMT-SCM, and it may also be the clinical spontaneity of CMT. The expression of healing.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R726.8

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