本文選題：MIP-1α + 趨化因子��； 參考：《重慶醫(yī)科大學(xué)》2008年碩士論文

【摘要】： 目的:創(chuàng)傷愈合是一個包括損傷組織炎癥反應(yīng),增生增殖和成熟階段的一個復(fù)雜生物學(xué)反應(yīng)。許多不同種類的生物物質(zhì)與創(chuàng)傷愈合的各個階段比如促進炎癥反應(yīng),肉芽組織的生成以及血管反應(yīng)等密切相關(guān)。在法醫(yī)病理學(xué)中,這些生物物質(zhì)在皮膚損傷中的表達可以用于損傷時間的推測。趨化性細胞因子又稱趨化因子,它們參與白細胞遷移、激活和趨化,在炎癥反應(yīng)中起核心作用。MIP-1α屬趨化因子CC亞族,分子量為8KD。MIP-1α可在不同因素刺激下被細菌,病毒及IL-1、TNF-α等炎性細胞因子誘導(dǎo)產(chǎn)生,據(jù)報道細胞來源的MIP-1α可由單核/巨噬細胞、中性粒細胞、成纖維細胞、內(nèi)皮細胞、及某些腫瘤細胞等多種細胞產(chǎn)生。MIP-1α的靶細胞有單核細胞、中性粒細胞、酸性粒細胞、NK細胞、樹突狀細胞等,其通過受體介導(dǎo)與靶細胞結(jié)合發(fā)揮生物學(xué)活性.MIP-1α與其受體結(jié)合后,有趨化免疫細胞、抑制造血干細胞生長及抑制HIV感染的作用。目前國內(nèi)外對MIP-1α研究多集中在臨床領(lǐng)域,而在法醫(yī)學(xué)領(lǐng)域的研究較少,尤其是國內(nèi)對MIP-1α在皮膚機械性損傷修復(fù)時間的推斷方面幾乎未見報道。本研究通過建立小鼠皮膚切創(chuàng)損傷模型,運用免疫組化技術(shù)及其陽性細胞計數(shù)法對損傷組織進行MIP-1α的檢測與分析,探討在皮膚創(chuàng)傷愈合過程中其表達的時序性變化規(guī)律,以便為法醫(yī)病理學(xué)損傷時間的推斷提供一個新的理論依據(jù)。 方法:采用完全隨機分組方法,分為實驗組和對照組。實驗組又分為生前造創(chuàng)組和死后造創(chuàng)組,生前造創(chuàng)組根據(jù)造創(chuàng)后處死時間不同將實驗小鼠隨機分為傷后1h、3h、6h、12h、1d、3d、6d、10d、14d共9組,死后造創(chuàng)組分為0.5 h,1 h,3 h,6h共4組;另設(shè)一組為正常對照組,即未造創(chuàng)組。生前造創(chuàng)組在小鼠背部制做全層切創(chuàng),在預(yù)定時間分別斷頸處死后即刻取材,死后造創(chuàng)組于處死小鼠后在預(yù)定時間造創(chuàng)取材,應(yīng)用免疫組織化學(xué)技術(shù)及陽性細胞計數(shù)法觀察傷后小鼠切創(chuàng)組織中不同時程的生前傷和死后傷皮膚創(chuàng)傷局部MIP-1α的表達,并和無切創(chuàng)的小鼠作為對照,探討MIP-1α在小鼠皮膚切創(chuàng)損傷愈合過程中的表達分布變化規(guī)律及其與損傷時間的關(guān)系. 結(jié)果:1、HE染色觀察:傷后1d內(nèi),可見大量炎性細胞游出,主要以中性粒細胞浸潤為主;傷后3～6d,肉芽組織逐漸形成并大量增生,炎性細胞以單核-巨噬細胞為主;傷后10～14d,肉芽組織逐漸成熟,炎性細胞消失,創(chuàng)口基本愈合。死后造創(chuàng)組及正常對照組鏡下無上述改變。2、免疫組化SABC法染色觀察:傷后1d內(nèi),MIP-1α在大量浸潤的中性粒細胞中呈陽性染色及在單核細胞中呈散在陽性表達;傷后3d,MIP-1α在逐漸形成并大量增生的肉芽組織中呈陽性染色,其主要在大量的單核細胞內(nèi)表達并在開始增生的成纖維細胞內(nèi)表達;其后到14d MIP-1α陽性表達在部分成纖維細胞內(nèi),但染色范圍和強度明顯減弱。死后造創(chuàng)組(0.5h,1h組)及正常對照組在部分表皮細胞及毛囊等表達弱陽性,死后造創(chuàng)3h,6h組表達陰性。3、陽性細胞計數(shù)法結(jié)果:陽性細胞率(PR值)在傷后6h到傷后3d逐漸上升,傷后3d達到高峰,傷后10～14d,PR值逐漸下降。4、統(tǒng)計學(xué)處理結(jié)果:方差分析結(jié)果表明生前創(chuàng)傷各相鄰兩組之間MIP-1αPR值相比均具有顯著差異(P0.01)。其中生前創(chuàng)傷3d組與其他各時間組相比均有顯著差異(P0.01)。 結(jié)論: 1、生前造創(chuàng)組MIP-1α在炎癥細胞中的表達量的變化具有規(guī)律性,呈現(xiàn)出升高期-高峰期-下降期的時序性變化曲線,其中造創(chuàng)后3d達到表達最高峰。2、MIP-1α表達陽性細胞的變化具有規(guī)律性,呈現(xiàn)出早期以中性粒細胞為主,高峰期以單核-巨噬細胞為主,后期以部分成纖維細胞表達為主的動態(tài)變化趨勢,死后造創(chuàng)組(0.5h,1h組)及正常對照組在部分表皮細胞及毛囊等表達。3、MIP-1α在小鼠皮膚切創(chuàng)愈合過程中呈現(xiàn)一定時序性變化,有望作為法醫(yī)損傷時間推斷的一個有用的參考指標,以及對于死后傷如1h以內(nèi)的早期損傷也有著重要的法醫(yī)學(xué)意義。
[Abstract]:Objective: wound healing is a complex biological response to the inflammatory response, proliferation and maturation of the injured tissue. Many different kinds of biological substances are closely related to the various stages of wound healing, such as the promotion of inflammation, the formation of granulation tissue, and the reaction of blood vessels. In forensic pathology, these biomarkers The expression of substance in skin damage can be used to speculate on the time of damage. Chemokines are also known as chemokines, which participate in leukocyte migration, activation and chemotaxis, and play a core role in the inflammatory response of.MIP-1 a CC subgroup of chemokine, and the molecular weight of 8KD.MIP-1 alpha can be affected by bacteria, viruses and IL-1, TNF- alpha under the same factors. Induced by sex cytokine, it is reported that the cell source MIP-1 alpha can be produced by mononuclear cells, neutrophils, acidic granulocytes, acid granulocytes, NK cells, dendritic cells and so on, which are mediated by the receptor and targeted by the target cells, which can be produced by a variety of cells, such as mononuclear / macrophage, neutrophils, fibroblasts, endothelial cells, and some tumor cells. Combining with biologically active.MIP-1 alpha and its receptor, cellular binding can chemotaxis immune cells, inhibit the growth of hematopoietic stem cells and inhibit the effect of HIV infection. At present, the study of MIP-1 alpha mainly focuses on the clinical field, but there are few studies in the field of forensic medicine, especially the time for the repair of MIP-1 alpha in the repair time of the mechanical damage of the skin. In this study, a mouse skin incision injury model was established, and the immunohistopathological technique and the positive cell count method were used to detect and analyze the MIP-1 alpha of the injured tissue, and the temporal variation of the expression during the healing process of skin wound was discussed in order to provide the inference for the time of forensic pathology injury. A new theoretical basis.
Methods: the experimental group and the control group were divided into the experimental group and the control group. The experimental group was divided into the pre birth and the postmortem group. The experimental mice were randomly divided into 9 groups, including 1H, 3h, 6h, 12h, 1D, 3D, 6D, 10d, 14d after the injury. The group was divided into 0.5 h, 1 h, 3 h, and 4 groups. In the normal control group, that is, the non creation group. The group was made in the back of the mice in the whole layer to cut the whole layer on the back of the mice, and immediately took the material after the death of the neck at the predetermined time. After death, the group was sacrificed at the predetermined time after the death of the mice. The immuno histochemical technique and the positive cell count were used to observe the different time injuries of the mice after the injury. The expression of local MIP-1 alpha in skin wounds after injury and injury was compared with that of noninvasive mice. The change of expression distribution of MIP-1 alpha in the healing process of skin incision injury in mice and its relationship with the time of injury were discussed.
Results: 1, HE staining observation: in 1D after injury, a large number of inflammatory cells were seen, mainly infiltration of inflammatory cells, mainly neutrophil infiltration, 3 to 6D after injury, granulation tissue gradually formed and proliferated, inflammatory cells were mainly mononuclear macrophages; 10 to 14d after injury, granulation tissue gradually mature, inflammatory cells disappeared, the wound was basically healed. Postmortem group and positive In the control group, there was no.2 and immunohistochemical SABC staining. In 1D, MIP-1 alpha was positive in a large number of infiltrating neutrophils and scattered in mononuclear cells, and 3D, MIP-1 alpha was positive after the injury, and it was mainly in a large number of mononuclear cells. Expression and expression in the fibroblasts which began to proliferate; then 14d MIP-1 alpha was positive in some fibroblasts, but the range and intensity of dyeing decreased obviously. After death, the expression of 0.5h, 1H group and normal control group were weakly positive in some epidermal cells and hair follicles. After death, 3h, 6h negative.3, positive cell count method were used. Results: the positive cell rate (PR) increased gradually from 6h to 3D after injury, and the 3D reached its peak after injury, 10 to 14d after injury, and the PR value gradually decreased by.4. The results of statistical analysis showed that the MIP-1 alpha PR values between the adjacent two groups were significantly different (P0.01). The preoperative trauma 3D group was compared with the other time groups. There were significant differences (P0.01).
Conclusion: 1, the changes in the expression of MIP-1 alpha in the inflammatory cells in the pre creation group are regular, showing a time series change curve of the peak period and the decline period, in which the 3D reached the peak of.2, and the change of the MIP-1 alpha expression positive cells is regular, and the early stage is mainly neutrophils, and the peak period is mononuclear at the peak period. - macrophage was the main trend of the expression of partial fibroblasts in the later period..3 was expressed in some epidermal cells and hair follicles in the postmortem group (0.5h, 1H group) and the normal control group. MIP-1 alpha showed certain temporal changes during the healing process of the skin incision in mice. It is expected to be a useful reference for forensic injury time. There is also an important forensic significance for the index and early injury of postmortem injury, such as 1H.

【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：D919

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