本文選題：局限性硬皮病　切入點：脂肪移植　出處：《北京協(xié)和醫(yī)學(xué)院》2017年博士論文

【摘要】：目的:分析脂肪移植治療局限性硬皮病的相關(guān)影響因素,探討脂肪干細胞(ADSCs)輔助脂肪移植治療局限性硬皮病的可行性。方法:(1)21例脂肪標本:局限性硬皮病患者脂肪標本6例(未使用藥物治療),局限性硬皮病患者脂肪標本5例(曾使用藥物治療),正常脂肪患者脂肪標本10例,提取脂肪標本中的干細胞,分離培養(yǎng),比較脂肪干細胞數(shù)量、形態(tài)、增殖能力及成脂能力。(2)激光散斑對比成像方法測定8例局限性硬皮病患者病變部位及正常對側(cè)部位的血流情況。(3)取局限性硬皮病病變部位皮膚標本2例及非硬皮病患者皮膚標本2例,行成纖維細胞培養(yǎng)后與脂肪顆�；靹蚝笠浦踩肼闶�,觀察脂肪的存活情況。(4)博來霉素硬皮病造模裸鼠18例,正常裸鼠18例,各分為三組:對照組,脂肪移植組,脂肪干細胞+脂肪移植組。術(shù)后1月取移植脂肪稱重,取注射部位裸鼠皮膚免疫組化測定TGF-β 1及Ⅲ型膠原含量。結(jié)果:(1)21例患者脂肪干細胞形態(tài)學(xué)無差異,成脂能力無差異,流式細胞儀檢測第3代患者脂肪干細胞的表面標志CD44+都是90%以上,無明顯差異。但局限性硬皮病患者(行激素治療)的干細胞數(shù)量較正常人及局限性硬皮病患者(未行激素治療)明顯減少,細胞生長曲線延遲。(2)局限性硬皮病患者病變部位血流灌注為92.69±22.04,正常對側(cè)部位為95.70 ±20.43,局限性硬皮病病變部位與對側(cè)正常部位血流灌注無明顯差異。(3)局限性硬皮病患者成纖維細胞+脂肪移植后脂肪重量約68.24±19.20mg,正常人成纖維細胞+脂肪移植后脂肪重量約88.42±20.15mg。(4)正常裸鼠脂肪移植組脂肪重量102.97±17.87mg,脂肪干細胞+脂肪移植組脂肪重量135.05 ±27.28 mg,硬皮病裸鼠移植脂肪重量77.54±12.40mg,脂肪干細胞+脂肪移植組脂肪重量102.48 ±23.16 mg。脂肪干細胞+脂肪移植的存活率均較單純的脂肪移植存活率高。(5)對于正常裸鼠,TGF-β 1含量對照組為939.34±216.92,脂肪移植組為974.98±470.18,脂肪干細胞+脂肪移植組為967.66±448.74,Ⅲ型膠原含量對照組為623.85±113.84,脂肪移植組為605.73 ±217.17,脂肪干細胞+脂肪移植為599.34±89.39。對于硬皮病裸鼠,TGF-β1含量對照組為1945.95 ±330.77,脂肪移植組為1418.86 ±376.82,脂肪干細胞+脂肪移植為1132.12±190.69,Ⅲ型膠原含量對照組為2629.39±746.62,脂肪移植組為1531.74±836.41,脂肪干細胞+脂肪移植為946.92±448.90。正常裸鼠經(jīng)脂肪移植或脂肪干細胞+脂肪移植后TGF-β 1含量及Ⅲ型膠原含量無明顯改變。但對于硬皮病裸鼠,脂肪移植或者脂肪干細胞+脂肪移植可降低硬皮病組裸鼠的TGF-β 1含量及Ⅲ型膠原含量,尤其是脂肪干細胞移植后效果更加明顯。結(jié)論:(1)局限性硬皮病患者的脂肪移植不同于正常人群的脂肪移植,長期應(yīng)用激素治療病史及病變部位的微環(huán)境是影響硬皮病患者脂肪移植存活的關(guān)鍵因素,而局限性硬皮病患者的病變部位血供較正常側(cè)無明顯改變。(2)脂肪干細胞輔助脂肪移植不僅可提高脂肪存活率,同時還可減輕硬皮病裸鼠的皮膚膠原沉積。
[Abstract]:Objective: to analyze the effect of fat transplantation for the treatment of localized scleroderma related factors, to explore the feasibility of adipose derived stem cells (ADSCs) transplantation for the treatment of localized scleroderma assisted fat. Methods: (1) 21 cases of fat samples: 6 cases of patients with localized scleroderma fat samples (without drugs use), 5 cases of fat were localized scleroderma patients (used drug treatment), 10 cases of patients with normal fat fat samples, extracted fat samples of stem cells, isolation and culture, compare adipose stem cell number, morphology, proliferation and adipogenic ability. (2) laser speckle contrast imaging method for the determination of 8 cases of patients with localized scleroderma lesions and normal contralateral parts of the blood flow. (3) the limited scleroderma skin lesions were 2 cases and non skin specimens of 2 cases of scleroderma patients, for fibroblast culture and fat particles after mixing transplanted into nude mice, observe the fat deposit Live. (4) bleomycin scleroderma model in nude mice in 18 cases, 18 cases of normal mice, were divided into three groups: control group, fat transplantation group, adipose derived stem cells + fat transplantation group. Postoperative January transplanted fat weight, take the injection site skin of nude mice was detected by immunohistochemistry and TGF- beta 1 type III collagen content. Results: (1) 21 cases of patients with adipose derived stem cells morphological differences, adipogenic ability difference, detection of third generation patients with adipose derived stem cells surface markers of CD44+ are more than 90% flow cytometry, no significant difference. But the patients with localized scleroderma (hormone treatment) the number of stem cells compared with normal people and patients with localized scleroderma (without hormonal treatment) was significantly reduced, the cell growth curve of delay. (2) patients with localized scleroderma lesions perfusion was 92.69 + 22.04, normal to the contralateral side was 95.70 + 20.43, blood flow of localized scleroderma lesions and contralateral normal area No significant differences in perfusion. (3) patients with localized scleroderma fibroblasts + fat transplantation fat weight about 68.24 + 19.20mg, normal human fibroblasts + fat transplantation fat weight of about 88.42 - 20.15mg. (4) normal nude mice transplantation group fat fat weight 102.97 + 17.87mg, fat stem cells + fat fat transplantation group the weight of 135.05 + 27.28 mg, scleroderma transplanted fat weight 77.54 + 12.40mg, the survival rate of adipose derived stem cells + fat transplantation group fat weight of 102.48 + 23.16 mg. + fat transplantation of adipose derived stem cells were relatively pure fat graft survival rate is high. (5) in normal mice, TGF- beta 1 in the control group was 939.34 + 216.92 that fat transplantation group is 974.98 + 470.18, fat stem cells + fat transplantation group is 967.66 + 448.74, type III collagen content of control group was 623.85 + 113.84, fat transplantation group is 605.73 + 217.17, + fat fat stem cell transplantation was 599.34 For scleroderma + 89.39. nude mice, TGF- beta 1 in the control group was 1945.95 + 330.77, fat transplantation group is 1418.86 + 376.82, + fat transplantation of adipose derived stem cells was 1132.12 + 190.69, type III collagen content of control group was 2629.39 + 746.62, fat transplantation group is 1531.74 + 836.41, fat stem cells + fat transplantation was 946.92. 448.90. normal nude mice by transplantation of adipose derived stem or fat content and collagen content of TGF- beta 1 type III cell transplantation + fat had no obvious change. But for scleroderma mice, fat transplantation or fat content and collagen content of 1 stem cell type III + fat transplantation can reduce the TGF- beta scleroderma mice, especially fat stem cell transplantation the effect is more obvious. Conclusion: (1) patients with localized scleroderma fat transplantation fat transplantation is different from the normal population, the micro environment of long-term use of hormone therapy and the history of the site of the lesion is affected in patients with scleroderma The key point of survival is fat transplantation. However, the blood supply of lesion sites of the patients with localized scleroderma is not significantly different from that of the normal side. (2) adipose tissue derived stem cells assisted fat transplantation can not only increase the fat survival rate, but also reduce the collagen deposition of the scleroderma nude mice.

【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：R593.25

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