間質(zhì)干細胞和急性肺損傷專業(yè)知識講座培訓課件

1、間質(zhì)干細胞和急性肺損傷專業(yè)知識講座間質(zhì)干細胞和急性肺損傷專業(yè)知識講座ARDS was first recognized in the 1960s1 as a clinical syndrome of severe acute respiratory failure presenting with hypoxemia and bilateral pulmonary infiltrates, most often in the setting of pneumonia, sepsis, or major trauma.ARDS最初在1960年，被人們認識為嚴重急性呼吸衰竭的臨床綜合征；血氧不足和雙

2、邊肺浸潤，通常繼發(fā)于肺炎、敗血癥、或嚴重的的創(chuàng)傷； 間質(zhì)干細胞和急性肺損傷專業(yè)知識講座2ARDS was first recognized in tdistinction between (ALI) and ARDS relates to the severity of hypoxemia, with the former having a PaO2/FiO2 of less than 300, the latter with a PaO2/FiO2 of less than 200. lung endothelial injury, alveolar epithelial injury, t

3、he accumulation of protein-rich fluid 、cellular debris in the alveolar space急性肺損傷(ALI)和ARDS的區(qū)別主要指血氧不足嚴重程度，前者有PaO2 /FIO2小于300;后者PaO2 /FIO2不足200;ALI / ARDS的發(fā)病機制涉及肺血管內(nèi)皮損傷, 肺泡上皮損傷,富含蛋白質(zhì)液體的增加和肺泡腔內(nèi)的細胞殘骸. 間質(zhì)干細胞和急性肺損傷專業(yè)知識講座3distinction between (ALI) and 流行病學2005年,大約200000名患者在美國發(fā)展為ALI / ARDS,估計有40%的死亡率。采用肺保護

4、性通氣時代,死亡率下降大約25%。 In 2005, approximately 200,000 pts in the United States ALI/ARDS, with an estimated mortality of 40%. In the era of lung protective ventilation, mortality has declined to approximately 25%.間質(zhì)干細胞和急性肺損傷專業(yè)知識講座4流行病學2005年,大約200000名患者在美國發(fā)展為間質(zhì)干大量的臨床試驗,藥物治療吸入表面活性劑（ inhaled surfactant ）、一氧化

5、氮（ nitric oxide ）、前列環(huán)素（ prostacyclins）,糖皮質(zhì)激素（ glucocorticoids）,抗氧化劑（antioxidants）, 酮康唑（ketoconazole）并非ALI治療標準； ALI肺泡的病理生理，任何單一分子不可能逆轉(zhuǎn)綜合癥的過程，使臨床獲益； reverse the course of this syndrome 、provide substantial clinical benefit. 間質(zhì)干細胞和急性肺損傷專業(yè)知識講座5大量的臨床試驗,藥物治療吸入表面活性劑（ inhaled s細胞為基礎的療法 cell-based therapies

6、能產(chǎn)生炎癥分子 molecules調(diào)節(jié)炎癥反應 modulate inflammatory cascades 增強修復 enhance repair 間充質(zhì)干細胞可能是理想的選擇 Mesenchymal Stem Cells（MSC） 間質(zhì)干細胞和急性肺損傷專業(yè)知識講座6細胞為基礎的療法 cell-based therapies 間充質(zhì)干細胞:一般屬性間充質(zhì)干細胞(msc) self-renewing isolated from bone marrow differentiate into muscle, bone, fat, fibroblasts, and cartilage. 能重建造血

7、的環(huán)境 reconstitute a hematopoietic environment , regenerate bone tissue；起初被稱為成纖維細胞的集落形成單位colonyforming unit-fibroblastic ，(CFU-F)；骨髓基質(zhì)細胞marrow stromal cells,MSC間質(zhì)干細胞和急性肺損傷專業(yè)知識講座7間充質(zhì)干細胞:一般屬性間充質(zhì)干細胞(msc) 間質(zhì)干細胞和急仍然不知道間充質(zhì)干細胞是否來自中胚層,或者神經(jīng)上皮,或來自不同來源的發(fā)展階段whether MSCs originate from the mesoderm, from the neuro

8、epithelium, or from different sourcesMSC 骨髓、脂肪,臍帶血,胎盤組織、肌腱和骨骼肌、 bone marrow 、also from fat, umbilical cord blood, placental tissue, tendons、skeletal muscle 間質(zhì)干細胞和急性肺損傷專業(yè)知識講座8仍然不知道間充質(zhì)干細胞是否來自中胚層,或者神經(jīng)上皮,或來自不國際上細胞治療提出標準：1.粘附整形adherence to plastic ；2. expression of CD105、CD73 CD90； lack of expression of

9、CD45、 CD34、CD14 CD11b,CD79a、CD19和人類白細胞抗原(HLA)II；3. ability to differentiate into osteoblasts, adipocytes, and chondroblasts in vitro. 成骨細胞、脂肪細胞、軟骨細胞的能力。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座9國際上細胞治療提出標準：間質(zhì)干細胞和急性肺損傷專業(yè)知識講座9other properties 1. produce a wide variety of molecules, including hematopoietic factors,chemokines,

10、 and angiogenic factors ,包括造血因子,趨化因子和血管生成素。2. immunomodulatory effects 3. MSC 基因治療的載體,并幫助誘導、同種異體骨髓移植耐受性。4. MSC在疾病潛伏期模型獲益，克羅恩病、創(chuàng)傷性腦損傷、Crohn disease、traumatic brain injury 。5.several types of lung disease 間質(zhì)干細胞和急性肺損傷專業(yè)知識講座10other properties 間質(zhì)干細胞和急性肺損傷專業(yè)MSCs IN LUNG DISEASE 博來霉素暴露小鼠肺纖維化、Bleomycin expo

11、sure fibrotic lung disease分離小鼠的干細胞并且靜脈注射，隨后7天，肺損傷的區(qū)域發(fā)現(xiàn)外源性間充質(zhì)干細胞存在，且細胞具有上皮細胞的特征；顯著降低肺內(nèi)膠原蛋白沉積，減少基質(zhì)蛋白酶2和9表達； reduced collagen deposition and expression of matrix metalloproteinases 2 and 9. 間質(zhì)干細胞和急性肺損傷專業(yè)知識講座11MSCs IN LUNG DISEASE 博來霉素暴露小鼠肺肺移植（ lung engraftment. ）：強大抗炎作用 antiinflammatory effects ；blocks

12、 increase in interleukin (IL)-1a，白介素1(IL)產(chǎn)生。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座12肺移植（ lung engraftment. ）：強大抗炎作green fluorescent protein (GFP) 標記MSC治療的博來霉素誘導6h的小鼠， improved survivalGFP標記的MSC，表達纖維細胞(fibroblasts),肌纖維細胞(myofibroblasts), type I and type II alveolar epithelial cells.間質(zhì)干細胞和急性肺損傷專業(yè)知識講座13green fluorescent pr

13、otein (GFP MSCs umbilical cord tissueintravenously bleomycin in mice； MSCs were identified 2 weeks inflamed portions of the lung。 臍帶組織干細胞，靜脈應用于老鼠體內(nèi)，2周內(nèi)肺內(nèi)發(fā)現(xiàn)。 MSC減少膠原蛋白及細胞信號轉(zhuǎn)導分子（Smad2）濃度，表明這些細胞已經(jīng)具有抗纖維化屬性。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座14 MSCs umbilical cord tissuebronchopulmonary dysplasia, MSCs intratracheally rat

14、s exposed to prolonged hyperoxia.reduced apoptosis, myeloperoxidase activity, and collagen deposition, inflammatory molecules IL-6, tumor necrosis factor (TNF)-a, differentiated into type II alveolar epithelial cells,支氣管肺的發(fā)育不良動物模型，長期氧過多暴露老鼠氣管內(nèi)注入干細胞，發(fā)現(xiàn)能夠顯著降低細胞凋亡、髓過氧化物酶活性和膠原沉積，以及炎性分子il - 6、腫瘤壞死因子(TNF)

15、。此外,少數(shù)細胞分化成II型肺泡上皮細胞。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座15bronchopulmonary dysplasia, M同樣試驗也提示可以提高存活率及運動耐量改善, 和減少肺泡和肺血管損傷,以及肺動脈高壓。類似試驗間充質(zhì)干細胞氣管內(nèi)給藥，能夠降低大鼠肺動脈高壓,改進血管內(nèi)皮功能。van Haaften and colleagues reported that intratracheal MSCs improved survival 、 exercise tolerance, and decreased alveolar and vascular lung injury, pu

16、lmonary hypertension間質(zhì)干細胞和急性肺損傷專業(yè)知識講座16同樣試驗也提示可以提高存活率及運動耐量改善, 和減少肺泡和肺rat model emphysema MSC reduced emphysematous changes.MSCs differentiated into type II alveolar epithelial cells. MSCs have therapeutic effects in several models of lung disease. antiinflammatory properties, mitigating the lung dam

17、age in ALI. 輻射引起的老鼠肺氣腫模型，間充質(zhì)干細胞減少肺氣腫形成，分化成II型肺泡上皮細胞；在肺部疾病的模型中有治療作用，適用于ALI。 間質(zhì)干細胞和急性肺損傷專業(yè)知識講座17rat model emphysema MSC reducMSCs IN ANIMAL MODELS OF ALI AND SEPSIS: EVIDENCE OF BENEFICIAL EFFECTS 干細胞在急性肺損傷和膿毒癥的動物模型：有利的證據(jù)脂多糖(LPS) 用于ALI的動物模型；氣道給予LPS，48h誘發(fā)中性粒細胞聚集，與微血管通透性增加有關(guān)；在ALI中，經(jīng)鼻腔注入LPS，MSC 4h后在外周血中顯

18、著增加。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座18MSCs IN ANIMAL MODELS OF They next subjected irradiated mice with bone marrow reconstituted from GFP transgenic donors to intranasal LPS, and showed abundant GFP-positive cells in the lungs 3 weeks later. Some of these cells expressed cytokeratin, a marker of epithelial cells,

19、 whereas others expressed CD34, a marker of endothelial cells.輻照骨髓抑制小鼠，骨髓重建鼻內(nèi)植入GFP標記MSC，3w在肺部發(fā)現(xiàn)GFP標記MCS。一些細胞表達上皮細胞標記細胞角蛋白（ cytokeratin）,而其他細胞表達CD34,內(nèi)皮細胞的標記。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座19They next subjected irradisuggested that endogenous MSCs play an important role in repairing inflammatory damage following LP

20、S.內(nèi)源性MSC在LPS誘導的急性炎癥反應修復中起重要的作用。在一些試驗中，低于致死劑量的輻照引起骨髓抑制，鼻腔內(nèi)的LPS產(chǎn)生類似的組織學損傷，且支氣管肺泡灌洗(BAL)中性細胞增多，1W后產(chǎn)生肺氣腫。表明缺乏內(nèi)源性MSC后破壞正常的修復過程。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座20suggested that endogenous MMSCs intravenously 30 minutes ， 3 days later in BAL total cell reduction decrease in inflammatory infiltrates, interalveolar septal

21、thickening, and interstitial edema. 氣管內(nèi)小鼠體內(nèi)，靜脈MSC 30min，3天可以發(fā)現(xiàn)顯著減少BAL中總細胞和中性粒細胞計數(shù)。 組織學分析證實減少炎癥浸潤,小葉間隔增厚,間質(zhì)水腫。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座21MSCs intravenously 30 minute intraperitoneal LPS 1 mg/kg (a dose that causes minimal mortality), 1 hour later infused MSCs or fibroblasts intravenously. MSCs, reduced lung

22、neutrophils at 6, 24, and 48 hours. 腹腔注射LPS 1mg/kg(最小致死量),1h后靜脈干細胞細胞或成纖維細胞干細胞治療6、24和48h后肺部N%及總數(shù)下降間質(zhì)干細胞和急性肺損傷專業(yè)知識講座22 intraperitoneal LPS 1MSC intratracheally to mice 4h，after 5 mg/kg LPS, （a dose significant mortality）. MSC-treated mice improved survival PBS-treated mice: 80% versus 42% 48 hours, 64

23、% versus 18% 72 hours. 間充質(zhì)干細胞小鼠氣管內(nèi)的給藥， 隨后4小時5毫克/公斤脂多糖氣管內(nèi)給藥，（可以產(chǎn)生顯著的死亡率劑量）干細胞治療后的小鼠改善生存相對于PBS-治療后老鼠:80%和42%在48小時內(nèi),80%對18%在72小時。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座23MSC intratracheally to mice 4Histologic analysis at 48 hours revealed less hemorrhage and edema. Nonviable MSCs and fibroblasts did not replicate this effe

24、ct, suggesting undifferentiated, viable MSCs were required to ameliorate LPS-induced ALI.MSC通過肺水（lung water）, 濕/干比（wet/dry ratio）, and BAL蛋白質(zhì)濃度 BAL protein concentration 48h后組織學顯示出血和水腫減少 提示未分化,有活性干細胞可以改善LPS誘導的ALI。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座24Histologic analysis at 48 hour在腸桿菌肺炎模型中 大腸桿菌誘導小鼠肺炎中。4h后,這些小鼠氣管內(nèi)注入MSC

25、、PBS,或成纖維細胞。通過BAL中性粒細胞計數(shù)衡量，干細胞顯著降低了肺部炎癥。MSCs, PBS, or fibroblasts. MSCs reduced lung inflammation, as measured by BAL neutrophil count.間質(zhì)干細胞和急性肺損傷專業(yè)知識講座25在腸桿菌肺炎模型中間質(zhì)干細胞和急性肺損傷專業(yè)知識講座25最近研究了盲腸的結(jié)扎和穿刺(CLP)膿毒癥小鼠模型。6h后，MSC或NS 靜脈注射，所有小鼠每天均有廣譜抗生素治療。Six hours following CLP, MSCs were infused intravenously. Al

26、l mice received daily broad-spectrum antibiotics. 間質(zhì)干細胞和急性肺損傷專業(yè)知識講座26最近研究了盲腸的結(jié)扎和穿刺(CLP)膿毒癥小鼠模型。間質(zhì)干細MSCtreated mice had decreased BAL cell counts and albumin、reduced inflammatory lung infiltrates and interstitial edema 28h. benefit of MSCs was not to the lungs, MSC-treated mice had reduced apoptotic

27、kidney cells and improved serum creatinine. CLP results in severe systemic injury, as shown by 45% mortality at 28h. MSC treatment improved mortality by 50%. 干細胞治療后小鼠肺泡灌洗液細胞計數(shù)和白蛋白減少。28h炎癥性肺部浸潤,間質(zhì)水腫減輕。除肺部外、腎臟細胞凋亡減少,血清肌酐下降。膿毒癥全身性損傷,28h 45%的死亡率。治療后死亡率下降50%。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座27MSCtreated mice had decreas

28、intravenous (IV) MSCs or control cells (heatkilled MSCs or skin fibroblasts) 24h before or 1 hour after CLP in mice. And broad-spectrum antibiotics. survival benefit for MSCs at 4 days. lower serum creatinine and kidney tubular injury scores, improved hepatic glycogen storage, and reduced transamina

29、ses, amylase, and splenic apoptosis,suggesting multiple beneficial systemic effects. IV MSC或控制細胞(熱死亡細胞或皮膚成纖維細胞)前24h或1h后膿毒癥小鼠模型中，廣譜抗生素。4d MSC治療生存率顯著提高，降低血清肌酐和腎小管損傷評分,改善肝臟糖原存儲,降低轉(zhuǎn)氨酶,淀粉酶、和脾細胞凋亡。多系統(tǒng)性有益。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座28intravenous (IV) MSCs or contrMSCs IN ANIMAL MODELS OF ALI AND SEPSIS: POTENTIAL ME

30、CHANISMS 干細胞在急性肺損傷和膿毒癥動物模型:潛在的機制MSCs may exert their therapeutic effects in models of ALI and sepsis, and have discovered an increasing number of potential mechanisms .It is helpful first to consider the processes governing the production and removal of alveolar edema fluid. 急性肺損傷和膿毒癥模型中, MSC發(fā)現(xiàn)越來越多的

31、潛在機制。首要原因為肺泡水腫液管理和去除。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座29MSCs IN ANIMAL MODELS OF ALI A內(nèi)皮滲透性、上皮通透性和肺泡液體間隙increased protein of the endothelial and epithelial barriers, and reduced (or insufficient) alveolar fluid clearance (AFC). MSCs reduce the increase in endothelial permeability ALI. BAL albumin and protein are co

32、mmonly used as markers of lung endothelial permeability。在肺泡水腫形成：內(nèi)皮蛋白質(zhì)通透性的增加；上皮屏障破壞，肺泡液體清除率下降。MSC能夠減少ALI內(nèi)皮通透性。BAL中白蛋白和蛋白質(zhì)含量通常用作的肺內(nèi)皮通透性指標。 間質(zhì)干細胞和急性肺損傷專業(yè)知識講座30內(nèi)皮滲透性、上皮通透性和肺泡液體間隙increased prBAL albumin, total protein, and immunoglobulin M (IgM) were increased 3 days following intratracheal LPS. This inc

33、rease was attenuated by MSCs given intravenously 30 minutes after the injury. Angiopoietin-1 may help maintain adultvascular endothelial cells in a quiescent state, reduce permeability and promote endothelial cell survival LPS注入氣管內(nèi)3天后肺泡灌洗液中白蛋白、總蛋白和免疫球蛋白M(IgM)增加。ALI靜脈注射MSC 30min，可以減輕蛋白含量。血管生成素維持血管內(nèi)皮細

34、胞處于靜止狀態(tài)，同事減少滲透性,促進內(nèi)皮細胞的存活。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座31BAL albumin, total protein, an MSCs produce Ang-1 further reduced BAL protein, albumin, and IgM 。 Ang-1 diminishing inflammatory cell influx and reducing plasma protein leakage into the alveolar space. 在未損傷老鼠中MSC促進血管生成素減少BAL蛋白,白蛋白,IgM的水平。推斷MSC靜脈注射入肺循環(huán)中，促使

35、Ang-1在減少炎性細胞浸潤,降低血漿蛋白滲漏。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座32 MSCs produce Ang-1 furth靜脈MSC，膿毒癥小鼠體內(nèi)28h可以減少肺泡灌洗液中白蛋白，提示MSC可以減少膿毒癥內(nèi)皮通透性老鼠給與大腸桿菌氣管內(nèi)注射，18h后肺泡灌洗液中白蛋白明顯增加，但是損傷后4h，氣管內(nèi)注射MSC(但不包括成纖維細胞)，白蛋白明顯減少。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座33靜脈MSC，膿毒癥小鼠體內(nèi)28h可以減少肺泡灌洗液中白蛋白，BAL中蛋白質(zhì)可以評估肺血管內(nèi)皮通透性、上皮通透性,肺水含量。 ALI中肺泡上皮細胞通常內(nèi)皮細胞屏障失去完整性。間質(zhì)干細胞和急性肺損傷專

36、業(yè)知識講座34BAL中蛋白質(zhì)可以評估肺血管內(nèi)皮通透性、上皮通透性,肺水含量 When exposed to inflammatory cytokines (IL-1b, TNF-a, and interferon IFN-g), increased 500%. MSCs protein reduced to control levels. 當暴露在強效的炎性細胞因子(IL-1b,TNF-a,和干擾素(IFN), 上皮細胞層蛋白質(zhì)滲透率增加大約500%。植入同種異體干細胞，蛋白質(zhì)滲透性減少到控制水平。ALI中，MSC可以提高上皮的屏障功能 間質(zhì)干細胞和急性肺損傷專業(yè)知識講座35 When exp

37、osed to inflammatoryAFC is the capacity of the epithelium of the lung to remove alveolar fluid。 ALI/ARDS can reduce AFC, including high tidal volume ventilation, live bacteria, acid instillation, and proinflammatory cytokines. 肺泡液體清除率（AFC），肺上皮細胞的清除任何原因肺泡液體,肺水腫能力。ALI / ARDS中通常是（AFC）受損與惡化的結(jié)果。影響AFC因素：高

38、潮氣量、細菌,和促炎細胞因子氣管內(nèi)注入MSC在ALI 4h后減少多余的肺水。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座36AFC is the capacity of the epi體外肺灌注的試驗，用來測試MSC在ALI中效果。 LPS decrease in AFC, approximately 20% per hour to near 0% per hour脂多糖導致肺泡液體清除率20%/h至0%/h。fibroblasts into the injured lung 1h after had no effect on AFC. MSCs AFC to baseline levels. 成纖維細

39、胞肺受傷1小時后應用，肺泡液體清除率沒有影響。干細胞肺泡液體清除率至基線水平。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座37體外肺灌注的試驗，用來測試MSC在ALI中效果。間質(zhì)干細胞和MSCs improve both endothelial/ epithelial permeability and AFC 。因此,得出結(jié)論干細胞似乎改善內(nèi)皮/上皮通透性和以及肺泡液體清除率。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座38MSCs improve both endothelial/移植Engraftment MSCs differentiate into bone, fat, muscle, cartilage

40、。in the late 1990s ， these cells could develop a nonmesodermal phenotype.MSC可以分化為骨,脂肪,肌肉,軟骨；1990年代末,在一定條件下,這些細胞可以發(fā)展為一個非中層表型。標記的干細胞融入博來霉素損傷后肺組織，表現(xiàn)出肺泡的二型肺細胞的形態(tài)學和分子特征。labeled MSCs incorporated into lung tissue bleomycin injury and developed molecular characteristics of type I pneumocytes. 間質(zhì)干細胞和急性肺損傷專業(yè)

41、知識講座39移植Engraftment MSCs differentilarge numbers of labeled MSCs in the lung 24h following IP LPS and IV MSC infusion.2 weeks later, few labeled cells remained, presence of MSCs in the lung is a transient in ALI. 靜脈注射脂多糖和靜脈注入干細胞24h后，大量標記MSC在肺內(nèi)發(fā)現(xiàn)。2周后,很少有標記的細胞存在,這表明大量存在的MSC在ALI中只是一種短暫的現(xiàn)象。間質(zhì)干細胞和急性肺損傷專業(yè)

42、知識講座40large numbers of labeled MSCs 對小鼠霧化脂多糖7天,第1天灌注入干細胞后，一些動物中,細胞可以表達Ang-1，輸液后14天,標記細胞出現(xiàn)在肺內(nèi)，但沒有表現(xiàn)進一步的特征 。 Taken together, suggest that MSC differentiation into mature lung cell types following ALI may occur at low levels but is unlikely to produce much of their beneficial effect. 綜上所述,數(shù)據(jù)表明,干細胞可以分化為

43、成熟的細胞，在ALI中可能發(fā)生在低水平,但不太可能產(chǎn)生的有益效果間質(zhì)干細胞和急性肺損傷專業(yè)知識講座41間質(zhì)干細胞和急性肺損傷專業(yè)知識講座41免疫調(diào)節(jié)（Immunomodulation ）MSCs were recognized to have immunomodulatory effects ，MSCs have suppress many functions of naive and memory T cells, B cells, natural killer (NK) cells, and the differentiation and function of monocytes MSC

44、抑制記憶T細胞、B細胞、自然殺傷(NK)細胞,單核細胞的分化和功能。膿毒癥模型中氣管內(nèi)的注入MSC可以使BAL中促炎細胞因子TNF-a和巨噬細胞炎性蛋白(MIP)2水平降低。 增加抗炎細胞因子il - 10、IL-1ra IL-13。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座42免疫調(diào)節(jié)（Immunomodulation ）MSCs we這些研究結(jié)果表明,在急性肺損傷中，干細胞可以促使肺環(huán)境從促炎到抗炎轉(zhuǎn)變。in a model of ALI shifts the injured lung from proinflammatory to antiinflammatory。靜脈注射MSC延緩脂多糖誘導的

45、ALI中血清促炎因子IFN-g， IL-1b ，MIP-1a,KC；膿毒癥小鼠動物模型中，可以靜脈注射MSC可以減少血清TNF-a和lL 6。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座43這些研究結(jié)果表明,在急性肺損傷中，干細胞可以促使肺環(huán)境從促炎MSC治療可以減少過氧物酶在脂多糖誘導小鼠膿毒癥的肝臟和腎臟中水平。MSC治療可以改善缺乏成熟的T細胞和B細胞基因敲除的小鼠，以及NK細胞缺陷老鼠的生存率。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座44MSC治療可以減少過氧物酶在脂多糖誘導小鼠膿毒癥的肝臟和腎臟在單核細胞和巨噬細胞缺陷小鼠模型中，發(fā)現(xiàn)MSC治療沒有效果, 表明這些細胞通過干細胞治療施加有益的影響。此

46、外MSC被證明增加環(huán)氧酶的表達和活性，增加前列腺素E2(PGE2)的水平。干細胞與巨噬細胞一起培養(yǎng),脂多糖模型中發(fā)現(xiàn)增加il 10，這種效果產(chǎn)生被證明依靠TLR4,MyD88 TLR4 NF-kB,TNF-a和TNF受體存在 間質(zhì)干細胞和急性肺損傷專業(yè)知識講座45在單核細胞和巨噬細胞缺陷小鼠模型中，發(fā)現(xiàn)MSC治療沒有效果,間質(zhì)干細胞和急性肺損傷專業(yè)知識講座培訓課件據(jù)報道, 脂多糖誘導膿毒癥24h后,小鼠靜脈治療，多能干細胞(傷后6小時)減少血清促炎的細胞因子,il - 6、IL-1b KC，和趨化因子配體5( CCL5)，血清il - 10也減少了,與原先實驗中這種現(xiàn)象相反 。尚不清楚為什么i

47、l 10在一個實驗中增加，在另一個實驗中減少,這可能與技術(shù)差異，MSC在損傷后應用的時間有關(guān)。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座47據(jù)報道, 脂多糖誘導膿毒癥24h后,小鼠靜脈治療，多能干細胞抗菌效果Antibacterial Effects MSC除了可以有效的抗炎效果，然而卻顯著改善由細菌引起的敗血癥小鼠生存率。實驗表明,MSC可以減少細菌的負擔能力。試驗發(fā)現(xiàn)，膿毒癥小鼠模型中靜脈注射脂多糖6h后，生存率極大改善。比較損傷后28h脾臟細菌菌落數(shù), MSC治療組菌落數(shù)下降一個數(shù)量級別 。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座48抗菌效果Antibacterial Effects MSC除在損傷后

48、24h，分離了小鼠腹膜內(nèi)和脾臟的總細胞數(shù)或CD11b1片段(單核細胞/巨噬細胞和中性粒細胞)。MSC治療組增加了總細胞和CD11b1革蘭氏陰性和革蘭氏陽性細菌的噬菌作用。MSC本身很少從事吞噬作用,必須間接調(diào)節(jié)宿主的吞噬細胞能力。 間質(zhì)干細胞和急性肺損傷專業(yè)知識講座49在損傷后24h，分離了小鼠腹膜內(nèi)和脾臟的總細胞數(shù)或CD11b其他實驗中，MSC同樣具有的抗菌性。干細胞(比較成纖維細胞)減少大腸桿菌體外的生長 。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座50其他實驗中，MSC同樣具有的抗菌性。間質(zhì)干細胞和急性肺損傷專MSC條件培養(yǎng)基對G陰性菌生長沒有影響,除非這些細胞原先被大腸桿菌細胞刺激,表明這種誘

49、導產(chǎn)生抗菌物質(zhì)。他們篩選細菌刺激的干細胞的培養(yǎng)基發(fā)現(xiàn)大量的人類(LL-37)。合成LL-37減少大腸桿菌和銅綠色假單胞菌的生長，當與LL-37抑制性抗體同時培養(yǎng)時,受刺激MSC失去了抗菌活性。老鼠膿毒癥模型中，MSC治療4h后,BAL顯示細菌減少超過一個數(shù)量級，而使用LL-37抗體，抗菌效果卻明顯下降。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座51MSC條件培養(yǎng)基對G陰性菌生長沒有影響,除非這些細胞原先被大Taken together, the results suggest that MSCs both exert direct effects on bacteria and modulate th

50、e hosts phagocytic capacity. Future experiments should help clarify the precise cellular and molecular pathways of phagocytic augmentation, and may identify additional direct antibacterial effects. 總的來說, 表明細胞對細菌起到直接的作用和調(diào)節(jié)宿主的噬菌作用的能力。未來的實驗應該進一步闡明的確切的細胞、分子途徑噬菌作用,和識別額外的直接的抗菌效果。間質(zhì)干細胞和急性肺損傷專業(yè)知識講座52Taken together, the results su其他可能的機制Other Possible Mechanisms MSC和組織細胞之間全新的互動機制。通過使用溴化乙錠改變DNA來減弱A549細胞線粒體功能。這些細胞同MSC一起培養(yǎng)后，A549細胞獲得了線粒體功能，且線粒體DNA同干細胞DNA相互匹配。顯微鏡檢提示MSC可以逐漸向A549細胞細胞質(zhì)擴展，并且通過這種擴展使線粒體大量轉(zhuǎn)運。間質(zhì)干細胞和急性肺損傷