本文選題：核心蛋白聚糖 + 創(chuàng)面愈合��； 參考：《蘭州大學》2015年碩士論文

【摘要】：目的：正常真皮組織中膠原纖維是有序的結構,由膠原蛋白分子高度有序的組裝構成。如果膠原蛋白排列、組裝和相互連接異常,則形成結構異常的膠原纖維支架,在創(chuàng)傷愈合后則表現(xiàn)為瘢痕形成。核心蛋白聚糖(DCN)對真皮創(chuàng)傷再生和瘢痕愈合中膠原纖維裝配影響的研究國內(nèi)很少。本研究通過建立兔耳皮膚創(chuàng)傷再生修復和瘢痕愈合一體化模型,于創(chuàng)面愈合過程在創(chuàng)周分別注射生理鹽水(NS),核心蛋白聚糖(DCN)及核心蛋白聚糖抗體(DCN抗體)。1.動態(tài)觀察并比較創(chuàng)面愈合過程中不同分組及不同深淺的組織形態(tài)學差異。2.觀察創(chuàng)面愈合過程中,Ⅰ型和Ⅲ型膠原蛋白在淺層和深層創(chuàng)面的表達差異。3.探討核心蛋白聚糖對創(chuàng)面愈合過程中Ⅰ、Ⅲ型膠原蛋白表達的影響。4.分析膠原纖維在淺層創(chuàng)面和深層創(chuàng)面愈合過程中的排列的差異,以及核心蛋白聚糖對其結構的影響。為進一步研究皮膚再生修復和瘢痕愈合提供形態(tài)學依據(jù)。方法：1.50只日本大耳白兔,按性別、年齡、體重相近的大白兔分配出25對,然后將每一對分別分配到不同的兩組中,分別為DCN組和DCN抗體組,每組25只,組內(nèi)設空白對照,視為正常對照(NS)組。于耳腹做大小為10.0mmx10.0mm由淺至深的斜行創(chuàng)面。傷后當日起,左耳創(chuàng)周以30G針頭注射濃度為5.0μg/ml的DCN(DCN組)或濃度為10.0μg/ml的DCN抗體(DCN抗體組),每次0.1ml,隔日一次,直至創(chuàng)面愈合；右耳注射等量生理鹽水作為對照(NS組),方法同上。所有創(chuàng)面均以無菌輔料包扎并卷耳固定。分別于傷后0天、4天、8天、12天及30天大體觀察和用標尺測量創(chuàng)面大小變化,并拍照存檔以便后期統(tǒng)計數(shù)據(jù)。比較各組平均愈合時間和觀察創(chuàng)面形態(tài)學改變。2.應用HE染色、免疫組化和計算機圖像處理法檢測日本大耳白兔耳腹側皮膚創(chuàng)傷模型外用DCN和DCN抗體前后,深淺創(chuàng)面愈合過程中的組織形態(tài)結構變化和Ⅰ、Ⅲ型膠原蛋白的表達差異。3.利用掃描電鏡技術觀測膠原纖維在深淺創(chuàng)面愈合過程中的變化規(guī)律,以及DCN對其結構的影響。結果：1.兔耳淺側創(chuàng)面(距淺側創(chuàng)面邊緣0-4.0mm內(nèi)為淺側創(chuàng)面)術后第8天,創(chuàng)面大部分結痂已脫離,再生修復。術后第30天,淺側色澤與正常皮膚無明顯差異,且界限不清,為再生愈合。深側創(chuàng)面(距淺側創(chuàng)緣內(nèi)約6-10mm為深側創(chuàng)面)直至術后第12天,創(chuàng)面仍見部分結痂存在,未完全上皮化,術后第30天,創(chuàng)面中央現(xiàn)小丘樣凸起,明顯高于皮面質(zhì)硬,色澤較周圍組織深,為瘢痕愈合。2.同一創(chuàng)面淺側或深側,DCN組創(chuàng)面較NS組創(chuàng)面愈合更快：創(chuàng)面DCN組創(chuàng)面愈合率第4天為(41.2±7.6)%、8天為(79.5±6.5)%、12天為(93.6±4.2)%,分別高于NS組的(36.6±4.3)%、(60.5±3.9)%和(85.2±4.3)%(P0.05)。應用DCN抗體后創(chuàng)面愈合率第4天(22.8±4.5)%、8天為(47.5±5.3)%、12天為(73.9±2.6)%,低于NS組(P0.05)；同時,DCN組愈合后組織色澤更接近正常組織,瘢痕組織更軟更薄,而DCN抗體組創(chuàng)面愈合后色澤深,瘢痕厚且硬,NS組介于兩者之間。3.創(chuàng)面干預后第30天時,DCN組再生面積(0.86±0.13)cm2高于NS組的(0.71±0.11)cm2(P0.05),而DCN抗體組再生面積(0.46±0.08)cm2低于NS組(P0.05)；DCN組瘢痕面積(0.14±0.07)cm2低于NS組的(0.29±0.09)cm2(P0.05),而DCN抗體組瘢痕面積(0.54±0.10)cm2高于NS組(P0.05)。4.創(chuàng)面Ⅰ、Ⅲ型膠原蛋白的表達：各個時間點Ⅰ、Ⅲ型膠原蛋白在淺側創(chuàng)面的表達均低于深側(P0.05)；各層次創(chuàng)面中Ⅰ膠原蛋白在各個時間點DCN組表達最低,DCN抗體組表達最高(P0.05),而Ⅲ型膠原蛋白在各組的表達無差異(P0.05)；各層次創(chuàng)面中Ⅲ/Ⅰ型膠原蛋白比例在各個時間點DCN組最高,DCN抗體組表達最低,差異明顯(P0.05)。5.創(chuàng)面膠原纖維的排列：各組淺側創(chuàng)面膠原纖維的排列在各個時間點均比深側創(chuàng)面更規(guī)律,增生不明顯,網(wǎng)狀結構更典型,纖維更細。同深淺側創(chuàng)面中,膠原纖維于各個時間點在DCN組排列更規(guī)律,纖維更細且增生不明顯,網(wǎng)狀結構更典型,而在DCN抗體組排列最紊亂,增生明顯,未見網(wǎng)狀結構且纖維最粗。結論：1.外源性DCN能調(diào)節(jié)Ⅲ/Ⅰ型膠原蛋白比例的作用,同時調(diào)節(jié)了膠原纖維的裝配,起到了預防瘢痕形成甚至實現(xiàn)再生修復的作用。2.本實驗成功建立了兔耳皮膚創(chuàng)傷再生和瘢痕愈合一體化模型,淺側創(chuàng)面實現(xiàn)了再生愈合,而深側創(chuàng)面實現(xiàn)了瘢痕愈合。
[Abstract]:Objective: the collagen fibers in normal dermal tissue are ordered structure, which are composed of collagen molecules highly ordered. If collagen is arranged, assembled, and interconnected, the collagen fibrous scaffold is formed with abnormal structure. After wound healing, it appears as scar formation. DCN There are few studies on the effect of collagen fiber assembly in scar healing. In this study, the integrated model of rabbit ear skin wound regeneration and scar healing was established, and the wound healing process was injected with physiological saline (NS), core proteoglycan (DCN) and core proteoglycan antibody (DCN antibody).1. in the wound healing process, and the wound healing was compared. Differences in histomorphological differences between different groups and different depths of the process.2. observation of the expression difference between type I and type III collagen in superficial and deep wounds during the wound healing process.3. probe into the effect of core proteoglycan on the expression of type I and III collagen in the wound healing process.4. analysis of the healing of collagen fibers in superficial and deep wounds The difference in the arrangement and the effect of the core proteoglycan on its structure. To provide morphological basis for further study of skin regeneration and scar healing. Methods: 1.50 Japanese white rabbits were assigned to 25 pairs by sex, age, and similar body weight, and each pair was assigned to two different groups, respectively. Group DCN and DCN antibody group, each group of 25, set blank control, considered as the normal control (NS) group. On the ear abdomen, the size of 10.0mmx10.0mm from shallow to deep of the oblique wound. From the day after injury, the left ear wound week with the 30G needle injection concentration of 5 mu DCN (DCN group) or the concentration of 10 micron DCN antibody (DCN antibody group), each time 0.1ml, straight every other day, straight To the wound healing, the right ear was injected with equal amount of saline as the control (group NS) and the method was same. All the wounds were wrapped with aseptic auxiliary materials and fixed with the ear. The size of the wound was measured at 0 days, 4 days, 8 days, 12 days and 30 days respectively. The morphological changes of the wound surface were observed by.2., HE staining, immunohistochemistry and computer image processing were used to detect the changes of tissue morphology and structure in the healing process of the deep and shallow wounds of the ears of Japanese big ear rabbits before and after the external use of DCN and DCN antibodies, and the difference of collagen type III with.3. was observed by scanning electron microscopy. The changes in the healing process of deep and shallow wounds and the effect of DCN on its structure. Results: 1. on the eighth day after the operation, most of the superficial wound of the rabbit ear (the edge of the superficial wound surface is the superficial wound in 0-4.0mm), most of the scab of the wound has been separated and regenerated. There is no obvious difference between the superficial color and the normal skin on the thirtieth day after the operation, and the boundary is not clear, for the regeneration of the skin. Healing. The deep side wound (about 6-10mm in the margin of the shallow margin) until twelfth days after the operation, the wound still found some scab, not completely epithelialization. On the thirtieth day after the operation, the center of the wound was a small hillock convex, obviously higher than the skin surface, and the color and lustre was deeper than the surrounding tissue. The wounds of the DCN group were wound in the superficial or deep side of the same wound, and the wound in group DCN was more than that of the NS group. The healing rate of the surface was faster: the wound healing rate in the wound DCN group was fourth days (41.2 + 7.6)%, 8 days (79.5 + 6.5)% and 12 days (93.6 + 4.2)%, respectively (36.6 + 4.3)%, (60.5 + 3.9)% and (93.6%)% (P0.05). The wound healing rate after the application of DCN antibody was (+ +)%, and was lower than that of the NS group (P0.05). At the same time, at the same time, After the healing of the DCN group, the tissue color was closer to the normal tissue, and the scar tissue was softer and thinner, while the wound healing was deep and the scar was thick and hard in the DCN antibody group. In the group NS, the regenerated area of the DCN group (0.86 + 0.13) cm2 was higher than that of the NS group (0.71 + 0.11) cm2 (P0.05), while the regeneration area of the DCN antibody group was (0.46 + 0.08) cm2. In group NS (P0.05), the scar area of group DCN (0.14 + 0.07) was lower than that of group NS (0.29 + 0.09) cm2 (P0.05), and the scar area of DCN antibody group (0.54 + 0.10) cm2 higher than that of NS group (P0.05).4. wound I, type III collagen expression: the expression of type III collagen egg white in the superficial wound was lower than that in the deep side. The expression of collagen in DCN group was the lowest at all time points, and the expression of DCN antibody group was the highest (P0.05), but there was no difference in the expression of type III collagen in each group (P0.05). The proportion of type III / I collagen in all layers was the highest in DCN group at every time point, and the expression of DCN antibody group was the lowest, and the difference was obvious (P0.05).5. wound collagen fiber arrangement: each group was in each group. The arrangement of collagen fibers in the superficial wound surface was more regular at each time point than the deep side wound. The proliferation was not obvious, the reticular structure was more typical and the fiber was finer. In the deep and shallow wounds, the collagen fibers were arranged more regularly in the DCN group at various time points, the fiber was finer and the proliferation was not obvious, the net structure was more typical, and the most disorder in the DCN antibody group was increased. Conclusion: 1. exogenous DCN can regulate the proportion of collagen type III / type I, adjust the assembly of collagen fiber and play the role of preventing scar formation and even realizing regenerative repair..2. experiment has successfully established the integrated model of wound regeneration and scar healing in the skin of rabbit ear skin. Wound healing was achieved and scar healing was achieved on deep side wounds.

【學位授予單位】：蘭州大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R622

【參考文獻】

相關期刊論文 前1條

1 孫其志;張穎;袁文;;Ⅵ型膠原蛋白的結構與功能以及與后縱韌帶骨化的關系[J];脊柱外科雜志;2010年05期

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