【摘要】：背景KM突變小鼠是我所KM封閉群小鼠繁殖過(guò)程中出現(xiàn)的一例自發(fā)突變小鼠,表型異常,全身被毛稀少,表皮增厚有皺褶。為進(jìn)一步育種研究,于SPF級(jí)動(dòng)物房?jī)?nèi)進(jìn)行近交化培育保種,其突變表型可穩(wěn)定遺傳。該KM突變小鼠初生時(shí)與野生型KM小鼠間表型無(wú)差異,至一周左右被毛生長(zhǎng)后即可與野生型相區(qū)分,其被毛稀疏,至離乳仍可見(jiàn)粉紅色皮膚,成年后皮膚明顯松弛褶皺,表面干燥,部分動(dòng)物有皮屑脫落,部分動(dòng)物出現(xiàn)皮膚潰破,類(lèi)似于人類(lèi)的慢性炎癥性皮膚病病變表現(xiàn)。因此我們對(duì)KM突變小鼠進(jìn)行近交系培育,以期發(fā)現(xiàn)其突變的遺傳規(guī)律并對(duì)突變基因進(jìn)行定位；同時(shí),研究中主要對(duì)該突變小鼠的生物學(xué)特性進(jìn)行觀察分析,檢測(cè)其在生物學(xué)特性方面與野生小鼠間的差異；根據(jù)突變小鼠的表型,觀察其皮膚的病理改變及免疫細(xì)胞和分子改變,檢測(cè)免疫器官脾臟和淋巴結(jié)淋巴細(xì)胞的改變等,目的在于了解KM突變小鼠的系統(tǒng)表型特點(diǎn)、遺傳規(guī)律等,為其后的基因定位和機(jī)制研究奠定基礎(chǔ),以期開(kāi)發(fā)新的疾病動(dòng)物模型。 方法選用同胞雜交、測(cè)交等方法近交培育的KM突變小鼠和對(duì)照野生小鼠作為研究對(duì)象,觀察其遺傳規(guī)律；電子攝像動(dòng)態(tài)觀察表型；對(duì)不同月齡,不同性別小鼠進(jìn)行體重、臟器系數(shù)、代謝率、體溫、及血液學(xué)常規(guī)、生化檢測(cè)；通過(guò)常規(guī)HE病理組織學(xué)、免疫組織化學(xué)及特殊染色對(duì)3月齡、6月齡KM突變小鼠和野生KM小鼠皮膚炎癥細(xì)胞及細(xì)胞因子進(jìn)行檢測(cè)比較及脂肪組織脂肪因子瘦素檢測(cè)比較；對(duì)不同年齡段的小鼠皮膚組織、脾臟及淋巴結(jié)細(xì)胞的凋亡與增殖以及脾臟、淋巴結(jié)組織T、B細(xì)胞數(shù)量進(jìn)行檢測(cè)比較。 結(jié)果KM突變小鼠近交培育至第10代,突變表型穩(wěn)定遺傳,通過(guò)觀察突變表型外顯率,基本符合孟德?tīng)柣蚍蛛x規(guī)律,且與性別無(wú)關(guān),推測(cè)為單基因控制的常染色體隱性遺傳突變；突變表型為皮膚稀毛、皮屑、皮皺等；KM突變小鼠的體重與同齡野生小鼠比較明顯降低,且生長(zhǎng)緩慢,但除胸腺及腹腔脂肪臟器系數(shù)比KM野生小鼠低之外,其它主要臟器重量系數(shù)均高于野生KM小鼠；體溫高,平均日飲水量大；外周血白細(xì)胞、淋巴細(xì)胞百分比比同齡野生小鼠多,粒細(xì)胞百分比隨年齡增長(zhǎng)下降；單核細(xì)胞百分比呈明顯增高的趨勢(shì)；血糖、總膽固醇、甘油三酯及高密度脂蛋白低、低密度脂蛋白高；皮膚組織病理表現(xiàn)為表皮細(xì)胞壞死,上皮角化過(guò)度或不全,顆粒層增厚,基底細(xì)胞層水腫,真皮淺層血管擴(kuò)張,結(jié)締組織炎細(xì)胞浸潤(rùn)等；皮膚真皮層T細(xì)胞、巨噬細(xì)胞、肥大細(xì)胞浸潤(rùn),炎癥因子IL-6、IL-22、TNF-α、IFN-γ表達(dá)增多；脂肪組織瘦素表達(dá)增多；表皮細(xì)胞凋亡增多,毛囊及皮脂腺細(xì)胞增殖減少；脾臟及淋巴結(jié)細(xì)胞凋亡相對(duì)增多,3月齡、6月齡KM突變小鼠增殖增加,9月齡突變小鼠脾臟增殖下降；3月齡、6月齡KM突變小鼠脾臟及淋巴結(jié)內(nèi)T、B淋巴細(xì)胞均相對(duì)增多,9月齡時(shí)脾臟及淋巴結(jié)B細(xì)胞下降,T細(xì)胞增多。 結(jié)論此次發(fā)現(xiàn)的KM自發(fā)突變小鼠突變表型主要為皮膚組織自發(fā)慢性炎癥病變,全身系統(tǒng)性炎癥改變,與人類(lèi)慢性炎癥性皮膚病變有相類(lèi)似的病理改變和細(xì)胞分子改變,且表型能穩(wěn)定遺傳。繼續(xù)深入研究并定位突變基因有望培育成為一種新的慢性炎癥性皮膚病的動(dòng)物模型。
[Abstract]:Background KM mutant mice are one of the spontaneous mutant mice in the breeding process of the KM closed group, and the phenotype is abnormal, the whole body is sparse, and the skin is thickened with wrinkles. In order to further study the breeding, in the SPF animal room, a close-crossing culture is carried out, and the mutant phenotype can be stably inherited. The KM mutant mouse has no difference between the phenotype of the mouse and the wild-type KM mouse, and can be distinguished from the wild-type after a week and so that the KM mutant mouse can be separated from the wild-type after a week, Some of the animals showed skin-breaking, similar to the human chronic inflammatory skin disease. In the same time, the biological characteristics of the mutant mice were observed and analyzed, and the difference between the biological characteristics and the wild mice was detected. according to the phenotype of the mutant mice, the pathological changes of the skin and the changes of the immune cells and the molecules are observed, the changes of the spleen and the lymph node lymphocytes of the immune organs are detected, and the like, Lays a foundation for subsequent gene positioning and mechanism research, with a view to developing new disease animal models. Methods KM mutant mice and control wild mice bred by the method of sibling hybridization and cross-crossing were used as the research object to observe the genetic law of the KM mutant mice and the control wild mice. The dynamic observation of the phenotype was observed. The body weight, the organ coefficient, the metabolic rate, the body weight, the body weight, the organ coefficient, the metabolic rate, the body weight, the body weight, the organ coefficient, the metabolic rate, the body weight, the body weight, the body weight, The results of routine HE, immunohistochemistry and special staining were used to detect the inflammatory cells and cytokines in the skin of three-month-old,6-month-old KM mutant mice and wild KM mice. The apoptosis and proliferation of mouse skin tissue, spleen and lymph node cells in different age groups and the number of T and B cells in the spleen and the lymph node were compared. Results KM mutant mice were bred to the 10th generation and the mutant phenotype was stable, and by observing the apparent rate of the mutant phenotype, it is basically in accordance with the Mendelian gene separation rule, and is not related to sex, and is presumed to be the autosomal recessive genetic mutation of single gene control; and the mutant phenotype is skin. The weight of KM mutant mice was significantly lower than that of the wild mice of the same age, but the weight coefficient of the other major organs was higher than that of the wild KM mice. Large amount of water; the percentage of white blood cells and lymphocytes in the peripheral blood is higher than that of the wild mice of the same age; the percentage of the granulocytes decreases with the age; the percentage of the monocytes is obviously increased; and the blood sugar, the total cholesterol, the triglyceride and the high-density lipoprotein are low, and the low-density lipoprotein high protein; that pathological manifestation of the skin tissue is epidermal cell necrosis, hyperkeratosis or incomplete, thickening of the particle layer, edema of the basal cell layer, superficial vascular expansion of the dermis, infiltration of connective tissue inflammatory cells, etc.; the skin dermal T cell, the macrophage, the mast cell infiltration, the inflammatory factor IL-6, IL, The expression of IL-22, TNF-1, IFN-antigen increased, the expression of leptin in adipose tissue increased, the apoptosis of the epidermal cells increased, the proliferation of the hair follicle and the sebaceous gland decreased, the apoptosis of the spleen and the lymph node was increased, the proliferation of the KM mutant in the 6-month-old and the 9-month-old mutant mice increased, and the spleen of the 9-month-old mutant mice increased. In the 3-month-old and 6-month-old KM mutant, the T and B lymphocytes in the spleen and the lymph nodes of the mice were increased, and the cells of the spleen and the lymph node B were decreased at the age of 9 months, and the T was fine. Conclusion The mutant phenotype of the KM spontaneous mutant mice is mainly the spontaneous and chronic inflammatory disease of the skin, the systemic inflammatory changes of the whole body, the pathological changes and the changes of the cell molecules, and the phenotype of the human chronic inflammatory skin. It can be stably inherited. It is expected that the mutation gene is expected to be cultivated as a new type of chronic inflammatory skin disease
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R-332

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本文編號(hào)：2455760