盐酸法舒地尔作用于兔耳增生性瘢痕的实验研究

发布时间：2018-05-20 03:16

本文选题：法舒地尔 + 增生性瘢痕　；参考：《泸州医学院》2012年硕士论文

【摘要】：目的：本实验通过对兔耳形成的增生性瘢痕内分别注射盐酸法舒地尔注射液（Fasudil Hydrochloride Injection，FHI）、醋酸曲安奈德注射液（TriamcinoloneAcetonaideAcetate Injection，TAAI）以及生理盐水（Normal Saline,NS），从大体及组织病理学的角度，对各组增生性斑块的瘢痕增生指数（Hypertrophic Index，HI）、成纤维细胞数量（Fibroblast Index，FBI）和对胶原纤维（Collagen fiber，CF）的形态、分布等各项指标进行比对和统计分析，研究盐酸法舒地尔注射液对增生性瘢痕组织的抑制作用。方法：健康标准化日本大耳兔12只，共24耳，雌雄各半，8-12月龄，体重2.0-2.5kg/只。氯胺酮1mg/kg肌注麻醉兔子后随机分为3组，每组4只，在兔耳建立增生性瘢痕模型，每只兔耳造模4个，总计96个。实验第28天随机抽取1例增生性斑块作病理切片染色证实增生性瘢痕形成，于每个增生性斑块内分别注射盐酸法舒地尔、醋酸曲安奈德注射液和生理盐水各30μl，分别于实验第28天、第35天、第42天对各组兔耳增生性瘢痕模型内进行注射，一共3次。观察并记录给药过程中瘢痕的形状、大小、质地。其后在实验第49天处死大耳兔，取兔耳增生性瘢痕组织的标本进行HE染色和Masson染色，光镜下观察瘢痕组织成纤维细胞和胶原纤维的形态、分布等，同时镜下通过测微尺计算各组标本中瘢痕增生指数，目测切片标本中成纤维细胞计数。实验结果均采用PASW Statistics18（SPSS18.0）统计软件对数据和图片进行统计学分析。结果：1、大体表现：生理盐水组（NS组）瘢痕仍明显挛缩并高于皮面，色深质硬，皮下可扪及明显结节，表面凹凸不平。盐酸法舒地尔组（FH组）和醋酸曲安奈德组（TAA组）瘢痕组织略高于皮面，未见明显挛缩，瘢痕颜色变浅，接近皮色，质地变软，皮下结节小，两组间瘢痕萎缩程度差异不明显。2、切片光镜下HE染色可见生理盐水组（NS组）毛细血管增生明显，成纤维细胞密集，核大深染，浅部呈漩涡状排列，深部则于软骨平行排列。胶原纤维排列杂乱，部分呈旋涡状；盐酸法舒地尔组（FH组）和醋酸曲安奈德组（TAA组）切片标本较生理盐水组（NS组）薄，表面平整，层次有序，可见毛细血管增生较生理盐水组（NS组）不显著，增生性组织内可见成纤维细胞和少量炎性细胞浸润，成纤维细胞细长。盐酸法舒地尔组（FH组）和醋酸曲安奈德组（TAA组）成纤维细胞胞核未见明显增大，细胞排列较紧密且比较有序，，数量较生理盐水组（NS组）减少，呈疏松状有序排列。胶原纤维较生理盐水组（NS组）规则排列。3、Masson染色下可见生理盐水组（NS组）胶原纤维蓝染较深，细胞间分布紊乱，生理盐水组（NS组）中粗大的胶原纤维蓝染较盐酸法舒地尔组（FH组）和醋酸曲安奈德组（TAA组）颜色更深，盐酸法舒地尔组（FH组）和醋酸曲安奈德组（TAA组）胶原纤维则淡染，纤细，疏松分布排列整齐。盐酸法舒地尔组（FH组）和醋酸曲安奈德组（TAA组）两组间比较胶原纤维形态、分布、染色等未见明显差异。4、生理盐水组（NS组）瘢痕增生指数（HI）以及成纤维细胞计数（FBI）均高于盐酸法舒地尔组（FH组）和醋酸曲安奈德组（TAA组），两两组间差异有统计学意义（P0.05）；盐酸法舒地尔组（FH组）和醋酸曲安奈德组（TAA组）两组间相比，盐酸法舒地尔组（FH组）和醋酸曲安奈德组（TAA组）瘢痕增生指数无显著性差异（P＞0.05），盐酸法舒地尔组（FH组）成纤维细胞计数高于醋酸曲安奈德组（TAA组），两两组间差异有统计学意义（p0.05），认为TAAI对成纤维细胞的抑制作用较FHI显著。结论：盐酸法舒地尔注射液（钙离子拮抗剂）及醋酸曲安奈德溶液（肾上腺皮质激素）对兔耳增生性瘢痕均有明显抑制作用，可见瘢痕组织变软、变平，醋酸曲安奈德溶液（TAAI）对成纤维细胞的抑制作用较盐酸法舒地尔注射液（FHI）更为明显，盐酸法舒地尔组（FH组）和醋酸曲安奈德组（TAA组）兔耳增生性瘢痕组织的瘢痕增生指数（HI）并无明显差异，二者对兔耳增生性瘢痕组织的胶原纤维无显著差异性影响。本实验认为，盐酸法舒地尔注射液（FHI）作为新型钙离子拮抗剂，可作为临床治疗增生性瘢痕的另一安全有效的治疗药物的选择，可为增生性瘢痕的治疗提供了一种新的思路和方法。
[Abstract]:Objective: in this experiment, Fasudil Hydrochloride Injection (Fasudil Hydrochloride Injection, FHI), Triamcinolone Acetonide Acetate Injection (TriamcinoloneAcetonaideAcetate Injection, TAAI) and physiological saline (Normal Saline, NS) were injected into the hypertrophic scar formed in the rabbit ear, and all the groups were increased from the general and histopathological point of view. The index of Hypertrophic Index (HI), the number of fibroblasts (Fibroblast Index, FBI) and the morphology and distribution of collagen fiber (Collagen fiber, CF) and other indexes were compared and statistically analyzed to study the inhibitory effect of Fasudil Hydrochloride Injection on hypertrophic scar tissue. Methods: healthy standardization of Japan. 12 rabbits with 24 ears, male and male, and male and male, and male and male, 8-12 months old, weight 2.0-2.5kg/ only. After anaesthetized rabbits with ketamine 1mg/kg intramuscular anaesthesia, 3 groups were randomly divided into 4 rats in each group. The model of hypertrophic scar was established in rabbit ears, 4 of each rabbit ear, total of 96. The experimental twenty-eighth days randomly selected 1 hypertrophic plaques for pathological section staining to prove hypertrophic scar formation, In each hyperplastic plaque, FFD, Triamcinolone Acetonide Acetate Injection and normal saline were 30 l each, respectively, in the twenty-eighth days, thirty-fifth days, and forty-second days of the rabbit ear hypertrophic scar model, respectively, 3 times. The shape, size and texture of the scar were observed and recorded in the forty-ninth days of the experiment. HE staining and Masson staining were taken for the specimens of hypertrophic scar tissue of rabbit ears. The morphology and distribution of fibroblasts and collagen fibers in scar tissue were observed under light microscope. At the same time, the index of hypertrophic scar hyperplasia was calculated by micrometer, and the count of fibroblast in the specimens of the specimens were counted. The results of the experiment were all PASW Stat Statistical analysis of istics18 (SPSS18.0) statistics software on the data and pictures. Results: 1, gross performance: the scar in the physiological saline group (group NS) is still markedly contracture and higher than the skin surface, the color is deep and hard, the subcutaneous can be palpable and the obvious nodule, the surface is uneven. The scar tissue of the famudil group (group FH) and the triamcinolone acetate group (group TAA) is slightly higher than the skin. Face, no obvious contracture, scar color light, close to skin color, texture soft, subcutaneous nodule small, the degree of scar atrophy between the two groups is not obvious difference between the two groups, HE staining under the microscope visible saline group (group NS) capillary hyperplasia, dense fibroblast cells, large deep dye, shallow vortex like arrangement, the deep cartilage in parallel arrangement. Glue. The primary fibers were arranged in a disorder and part of the whirlpool. The sliced specimens of the fasudil group (group FH) and triamcinolone acetate (group TAA) acetate (group NS) were thinner than the saline group (group NS), the surface was flat and orderly, and the capillary hyperplasia was not significant in the saline group (group NS), and the fibroblasts and a small amount of inflammatory cells infiltrated in the proliferative tissue. Fibrous cells were slender. The nuclei of fibroblasts in group FH and triamcinolone acetate (group TAA) were not significantly increased, and the cells were arranged closely and orderly. The number of cells in the saline group (group NS) decreased and arranged in a loose and orderly arrangement. The collagen fibers were arranged in a regular arrangement of.3 in the saline group (group NS), and the physiology was observed under Masson staining. The collagen fibers in the saline group (group NS) were dyed deeply, and the distribution of the cells was disordered. The thick collagen fiber blue dye in the saline group (group NS) was more darker than the FFD group (group FH) and the triamcinolone acetate group (group TAA). The collagen fibers in the famudil group (group FH) and the triamcinolone acetate group (group TAA) were pale, thin and loosely distributed. There was no significant difference in collagen fiber morphology, distribution and staining between the two groups (group FH) and triamcinolone acetate group (group TAA). The scar proliferation index (HI) and fibroblast count (FBI) in the saline group (group NS) were higher than that of the FFD group (group FH) and the triamcinolone acetate group (TAA group), and the difference between the 22 groups was poor. There was no statistically significant difference (P0.05); there was no significant difference between the two groups (group FH) and triamcinolone acetate (group TAA) group (group FH) and triamcinolone acetate group (group TAA) with no significant difference (P > 0.05), and the count of fibroblasts in group FH (group FH) was higher than that of triamcinolone acetate (TAA group). The difference between the 22 groups was statistically significant (P0.05), and the inhibitory effect of TAAI on fibroblasts was more significant than that of FHI. Conclusion: Fasudil Hydrochloride Injection (calcium antagonist) and triamcinolone acetate solution (adrenocortical hormone) have obvious inhibitory effect on hypertrophic scar of rabbit ears. It can be seen that the scar tissue softened, leveled, and acetamide acetate. The inhibitory effect of TAAI on fibroblasts was more obvious than that of Fasudil Hydrochloride Injection (FHI). There was no significant difference in scar proliferation index (HI) in the hypertrophic scar tissue of rabbit ear hydrochloric acid group (group FH) and triamcinolone acetate (TAA group). There was no significant difference between the two of the collagen fibers in the hypertrophic scar tissue of rabbit ears. This experiment suggests that Fasudil Hydrochloride Injection (FHI), as a new type of calcium ion antagonist, can be used as another safe and effective therapeutic drug for clinical treatment of hypertrophic scar. It can provide a new way of thinking and method for the treatment of hypertrophic scar.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R965;R758.6

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