發(fā)布時間：2018-11-24 16:14

【摘要】：【目的】 將銀屑病患者骨髓間充質(zhì)干細胞(Bone marrow Mesenchymal Stem Cells,BMMSCs)體外分化為血管內(nèi)皮細胞(VEC),并對定向分化的VEC活性進行初步研究,為銀屑病患者BMMSCs生物特性的深入研究提供依據(jù)。 【方法】 ①采用密度梯度離心法分離銀屑病患者與對照組骨髓單個核細胞,通過貼壁法培養(yǎng)BMMSCs,用流式細胞儀鑒定細胞表面標志;②加入血管內(nèi)皮細胞生長因子(VEGF)、堿性成纖維細胞生長因子(bFGF)定向誘導BMMSCs向VEC分化,流式細胞儀進行細胞表型鑒定;③免疫細胞熒光技術(shù)鑒定VEC標志物KDR、VWF的表達;④體外血管成形試驗檢測誘導的內(nèi)皮細胞體外成形能力;⑤用transwell小室測誘導的VEC對T細胞的遷移能力。⑥并對在培養(yǎng)分化過程中,發(fā)現(xiàn)1例雙親均患病的銀屑病患者BMMSCs在原代即開始向VEC分化,采用了高通量RNA測序技術(shù)對其差異表達基因進行了篩選。 【結(jié)果】 ①流式測定顯示兩組培養(yǎng)的BMMSCs高表達CD44、CD29,而HLA-DR、CD45、CD34陰性表達,說明所培養(yǎng)的BMMSCs純度較高;②銀屑病組CD34、CD45、ICAM-1、HLA-R陽性表達,且明顯高于對照組(p㩳0.05);③免疫細胞化學顯示兩組都可分化為VEC;④銀屑病患者誘導的VEC具有體外成形能力;⑤銀屑病組誘導的VEC對T細胞的具有遷移作用;⑥1例自分化患者高通量測定結(jié)果,共篩選了475個差異表達基因,其中298個基因表達呈增高趨勢,177個基因呈降低趨勢,其中,差異表達基因在2個term上有顯著富集(P0.05),分別為類前列腺素的代謝過程和前列腺素的代謝過程。 【結(jié)論】 銀屑病患者BMMSCs生物學活性異常,且易于轉(zhuǎn)分化為VEC。銀屑病患者BMMSCs參與了銀屑病的發(fā)病,其向VEC的異常分化可能是導致銀屑病患者皮損真皮乳頭層微血管異常增生的根源。
[Abstract]:[objective] to differentiate bone marrow mesenchymal stem cells (Bone marrow Mesenchymal Stem Cells,BMMSCs) from psoriatic patients into vascular endothelial cells (VEC),) in vitro and to investigate the activity of VEC in vitro. To provide evidence for further study of biological characteristics of BMMSCs in patients with psoriasis. [methods] 1Bone marrow mononuclear cells were isolated from psoriatic patients and control group by density gradient centrifugation. BMMSCs, was cultured by adherent method and cell surface markers were identified by flow cytometry. 2Vascular endothelial growth factor (VEGF),) basic fibroblast growth factor (bFGF) was added to induce the differentiation of BMMSCs into VEC, flow cytometry was used to identify the cell phenotype, and the expression of VEC marker KDR,VWF was identified by immunocytofluorescence technique. (4) the ability of endothelial cells to be formed in vitro was detected by in vitro angioplasty test. (5) the ability of T cell migration induced by VEC was measured by transwell chamber. 6 during the course of culture and differentiation, it was found that BMMSCs of one psoriatic patient with disease of both parents began to differentiate into VEC in primary culture. The differentially expressed genes were screened by high throughput RNA sequencing. [results] 1Flow flow analysis showed that the BMMSCs of the two groups was highly expressed CD44,CD29, and HLA-DR,CD45,CD34 negative, indicating that the purity of the cultured BMMSCs was high. (2) the positive expression of CD34,CD45,ICAM-1,HLA-R in psoriasis group was significantly higher than that in control group (p0. 05). (5) VEC induced by psoriasis could induce the migration of T cells; In 61 patients with self-differentiation, 475 differentially expressed genes were screened, 298 of which showed an increasing trend of expression and 177 of which showed a decreasing trend. The differentially expressed genes were significantly enriched on two term (P0.05), which were the metabolic process of prostaglandin-like and prostaglandin respectively. [conclusion] the biological activity of BMMSCs in psoriatic patients is abnormal, and it is easy to differentiate into VEC.. BMMSCs is involved in the pathogenesis of psoriasis and its abnormal differentiation to VEC may be the cause of abnormal proliferation of dermal papillary layer in psoriatic patients.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R758.63

【參考文獻】

相關期刊論文 前6條

1 張開明,李新華;骨髓,銀屑病的發(fā)病“中樞”?[J];中國皮膚性病學雜志;2004年08期

2 尹國華;李新華;張開明;劉玉峰;;銀屑病患者骨髓CD34~+細胞體外定向分化的T細胞活性研究[J];中華皮膚科雜志;2006年03期

3 牛旭平;徐海濤;尹國華;張開明;李新華;;銀屑病患者外周血單一核細胞的培養(yǎng)上清液對骨髓造血干細胞和祖細胞集落形成的影響[J];中華皮膚科雜志;2006年03期

4 蔡綏R，

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