经不同途径移植人骨髓间充质干细胞对热烟雾吸入性肺损伤大鼠的治疗作用

发布时间：2018-03-07 16:25

本文选题：热烟雾吸入性肺损伤　切入点：hMSCs　出处：《辽宁医学院》2012年硕士论文　论文类型：学位论文

【摘要】：目的 1、探讨经气管移植人的骨髓间充质干细胞对热烟雾吸入性肺损伤大鼠的作用； 2、探讨经尾静脉注入人骨髓间充质干细胞对热烟雾吸入性肺损伤大鼠的作用； 3、研究不同移植途径对干细胞发挥作用的影响。方法 1、自行研制热烟雾发生装置，模拟火灾现场热烟雾环境。将56只雄性健康Wistar大鼠随机数字表法分为正常对照组(8只)、热烟雾吸入致伤组(24只)和经气管移植hMSCs治疗组（24只），后两组再按照伤后时间点分为3个亚组（2h、4h、8h）。对照组在8h之后活杀，热烟雾吸入致伤组和经气管移植hMSCs治疗组分别于致伤后2h,4h,8h活杀。检测血清及支气管肺泡灌洗液（BronchoalveolarLavage Fluid BALF）中肿瘤坏死因子-α（Tumor Necrosis Factor-α TNF-α）、白介素-6（Interleukin-6,IL-6）和白介素-10（Interleukin-10，IL-10）的浓度。计算肺组织含水量湿重/干重比(Wet/Dry ratio W/D)；观察比较各组动物肺组织及气管标本大体改变、HE染色常规病理学改变。 2、48只健康Wistar大鼠同样按照上述方法制作热烟吸入性损伤动物模型，随机数字表法分为热烟雾吸入致伤组(24只)和经尾静脉移植hMSCs治疗组（24只），各组再按照致伤后时间点分为3个亚组（2h、4h、8h）。热烟雾吸入致伤组和经尾静脉移植hMSCs治疗组分别于致伤后2h,4h,8h活杀。测定血清及支气管肺泡灌洗液（BALF）中肿瘤坏死因子-α（TNF-α）浓度；白介素-6（,IL-6）和白介素-10（IL-10）的含量。计算肺组织含水量湿重/干重比(W/D)；观察比较各组动物肺组织及气管标本大体改变、HE染色常规病理学改变。结果 1、与正常对照组（C组）比较，热烟雾吸入性肺损伤组（SQ组）各时间点血清促炎、抗炎因子均显著升高；各时间点支气管肺泡灌洗液促炎因子显著升高，抗炎因子无明显变化。与SQ组比较，经气管移植hMSCs治疗组（MQ组）在4h、8h血清促炎因子显著下降，抗炎因子显著升高，在2h时炎性因子无明显变化。各时间点肺泡灌洗液促炎因子显著降低，而抗炎因子仅在4h、8h显著升高。与C组比较，SQ组和MQ组肺水质量分数均明显升高，MQ组肺水质量分数较SQ组显著降低；大体观察和组织病理学显示：MQ组肺组织损伤程度较SQ组明显改善； 2、与正常对照组（C组）比较，热烟雾吸入性肺损伤组（SV组）各时间点血清促炎、抗炎因子均显著升高；各时间点肺泡灌洗液促炎因子显著升高，，抗炎因子无明显变化。与SV组比较，经尾静脉注入hMSCs治疗组（MV组）各时间点血清促炎因子显著下降，抗炎因子显著升高。各时间点肺泡灌洗液促炎因子显著降低，而抗炎因子仅在4h、8h显著升高。与C组比较，SV组和MV组肺水质量分数均明显升高，MV组肺水质量分数较SV组显著降低；大体观察和组织病理学显示：MV组肺组织损伤程度较SV组明显改善。结论 1、经气管移植hMSCs后对热烟雾导致的急性肺损伤有一定的保护作用。 2、经尾静脉注入hMSCs能降低热烟雾吸入性肺损伤早期促炎因子水平，升高抗炎因子水平，减轻全身炎症反应，对损伤肺组织具有一定的保护作用。 3、干细胞经这两种移植途径均可对损伤肺组织发挥保护作用。
[Abstract]:objective
1, the effect of bone marrow mesenchymal stem cells (MSCs) on heat smoke inhalation induced lung injury in rats was investigated.
2, the effect of human bone marrow mesenchymal stem cells injected into the tail vein on hot smoke inhalation lung injury in rats was investigated.
3, the effect of different transplantation approaches on stem cells was studied.
Method
1, developed hot smoke generating device, simulating fire scene hot smoke environment. 56 healthy male Wistar rats were randomly divided into normal control group (8 rats), injury group (24 rats) inhalation of hot smoke and trachea transplantation hMSCs treatment group (24), after the two groups according to the time points after injury were divided into 3 subgroups (2h, 4h, 8h). The control group after 8h killed, hot smoke inhalation injury group and hMSCs treatment group were transplanted trachea after injury in 2H, 4h, 8h were killed. The serum and bronchoalveolar lavage fluid (BronchoalveolarLavage Fluid BALF) in tumor necrosis factor alpha (Tumor alpha TNF- alpha Necrosis Factor-), interleukin -6 (Interleukin-6, IL-6) and interleukin -10 (Interleukin-10, IL-10). The concentration of calculating the water content of lung tissue wet weight / dry weight ratio (Wet/Dry ratio W/D); observed in lung tissue of animal and gas samples general HE staining pathological change Learn to change.
2,48 healthy Wistar rats were also produced in accordance with the above method of hot smoke inhalation injury animal model, randomly divided into hot smoke inhalation injury group (24 rats) and intravenous transplantation of hMSCs treatment group (24 rats), each group according to the time point after injury were divided into 3 subgroups (2h, 4h, 8h). The injury group and intravenous transplantation of hMSCs treatment group respectively at 2h after injury, inhalation of hot smoke 4H killed 8h. Determination of serum and bronchoalveolar lavage fluid (BALF) in tumor necrosis factor alpha (TNF- alpha) concentration; interleukin (-6, IL-6) and white interleukin -10 (IL-10). The contents of calculating the water content of lung tissue wet weight / dry weight ratio (W/D); observed lung and trachea animal specimens in general change, HE staining pathological changes.
Result
1, with the normal control group (group C), hot smoke inhalation injury group (SQ group) each time point serum proinflammatory and anti-inflammatory cytokines were significantly increased at each time point; bronchoalveolar lavage inflammatory cytokines increased significantly, anti-inflammatory factor has no obvious change. Compared with SQ group, trachea transplantation hMSCs the treatment group (MQ group) in 4h, 8h of serum proinflammatory cytokines decreased significantly, anti-inflammatory factor increased significantly in 2H and inflammatory cytokines had no obvious change. At each time point in bronchoalveolar lavage fluid of pro-inflammatory cytokines and anti-inflammatory cytokines decreased significantly only in 4h, 8h was significantly increased. Compared with C group, SQ group and MQ group of lung water content were significantly increased in MQ group, lung water mass fraction was significantly lower than that of SQ group; morphology and histopathology showed that the degree of lung injury in group MQ was significantly better than those in SQ group;
2, with the normal control group (group C), hot smoke inhalation injury group (SV group) each time point serum proinflammatory and anti-inflammatory cytokines were significantly increased at each time point; bronchoalveolar lavage fluid of proinflammatory cytokines increased significantly, anti-inflammatory factor has no obvious change. Compared with SV group, intravenous injection hMSCs treatment group (group MV) at different time points of serum proinflammatory cytokines decreased significantly, anti-inflammatory factor significantly increased. At each time point in bronchoalveolar lavage fluid of pro-inflammatory cytokines and anti-inflammatory cytokines decreased significantly only in 4h, 8h was significantly increased. Compared with C group, SV group and MV group of lung water content were significantly increased, MV group of lung water content was significantly lower than that of SV group; morphology and histopathology showed: in MV group the degree of lung injury was significantly better than those in SV group.
conclusion
1, lead to acute lung heat smoke trachea transplantation hMSCs after injury has a protective effect.
2, injection of hMSCs through the caudal vein can reduce the level of proinflammatory factors in early stage, increase the level of anti-inflammatory factors, relieve systemic inflammatory response and protect lung tissue.
3, the stem cells can play a protective role in the injury of lung tissue through these two kinds of transplantation.

【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R563.8

【参考文献】