支气管哮喘是由嗜酸粒细胞、 肥大细胞和T淋巴细胞等多种细胞和细胞组分参与的气道慢性炎症性疾患，主要病理特点为上皮层大量的嗜酸粒细胞浸润及以上皮下纤维化、平滑肌增生、胶原蛋白沉积为主的气道重塑。哮喘的发病机制较为复杂，而炎症因子表达异常在哮喘的发病中发挥主要作用[1-3]。哮喘是一种全球范围内的常见病、多发病，我国约有1000万以上哮喘患者，而目前对于哮喘尚缺乏有效的根治方法。 间充质干细胞（mesenchymal stem cells，MSC）是具有强大的增殖能力和多向分化潜能的成体干细胞，同时具有免疫调节作用，它能通过免疫调节作用改善多种免疫相关性疾病的病情，而既往MSC在呼吸系统疾病中的研究主要集中在急性肺损伤，在哮喘当中的研究甚少。对于哮喘这一类以炎症因子表达异常为主的变态反应性疾病，MSC是否可以用于哮喘的治疗，值得我们进一步探讨。
ObjectiveTo study immunodepression effect of bone marrow-derived mesenchymal stem cell (BMSC) on acute asthmatic airway inflammation by galectin-1 (gal-1) in vivo.MethodsEighty-five female BALB/c mice were equally randomized into normal control group, asthmatic group, BMSC treatment group, gal-1 treatment group and BMSC and gal-1 inhibitor group. Ovalbumin (OVA) was used to establish acute asthmatic model. Total cell number and differential cell analysis in each group in bronchoalveolar lavage fluid (BALF) were determined. Furthermore, hematoxylin-eosin and periodic-acid Schiff staining was used to compare airway inflammation among five groups. Measurement of cytokines, including interleukin (IL) -4, IL-5 and gal-1 in BALF and OVA specific IgE (OVA-IgE) in serum were evaluated by enzyme linked immunosorbent assay. Moreover, dendritic cell (DC) in lung tissue was sorted by immunomagnetic beads and its MAPK signal pathway was analyzed by western blotting among five groups.ResultsAccumulation of inflammation cells, particularly eosinophils around airway and in BALF was evident in asthmatic mouse model, meanwhile hyperplasia of Goblet cell was also obvious in asthmatic group. BMSC engraftment or gal-1 infusion significantly reduced airway inflammation and hyperplasia of Goblet cell and the number of inflammation cells in BALF, especially eosinophils attenuated dramatically. However, there was no effect on airway inflammation and hyperplasia of Goblet Cell by simultaneous infusion BMSC engraftment and gal-1 inhibitor. Compared to normal control group, the level of IL-4, IL-5 in BALF and OVA-IgE in serum was increased remarkably in asthmatic group, but the level of gal-1 reduced obviously. Moreover, infusion of BMSC or gal-1 could mitigate the level of IL-4, IL-5 in BALF and OVA-IgE in serum and increase the level of gal-1 in asthmatic mouse. However, infusion with both BMSC and gal-1 inhibitor exerted no effect on cytokine and OVA-IgE in asthmatic mouse. DC was sorted by immunomagnetic beads and western blotting was used to detect the expression of MAPK signal pathway among five groups. The expression of ERK phosphorylation in asthmatic group was much lower than that in normal control group. On the contrary, the expression of p38 phosphorylation was much higher than that in normal control group. BMSC engraftment or gal-1 infusion significantly activated the ERK pathway and inhibited the p38 MARP pathway on asthmatic mouse DC. Nevertheless, the expression of ERK phosphorylation and p38 phosphorylation for group with BMSC and gal-1 inhibitor infusion was between the level of asthmatic group and normal control group.ConclusionsBMSC infusion alleviates airway inflammation in asthmatic mouse, especially weakens eosinophils infiltration, and the underlying mechanism might be protective effect of gal-1 secreted by BMSC which plays a role in lung tissue DC and regulates the DC expression of MAPK signal pathway.
【Abstract】 Objective To investigate the effect of allogeneic bone marrow-derived mesenchymal stem cells ( BMSCs) transplantation on the airway inflammation and airway remodeling in chronic asthmatic mice. Methods Forty female BALB/c mice were equally randomized into four groups, ie. a normal control group, a BMSCs control group, an asthma model group, and a BMSCs transplantation group. BMSCs were generated from male donor mice, then the mice in the asthma model group and the BMSCs transplantation group were sensitized and challenged with OVA to establish chronic asthmatic mice model. Hematoxylin and eosin staining and Alcian blue-periodic acid-Schiff staining were used to analyze the effects on airway inflammation and airway remodeling after BMSC engraftment. The number of CD4 + CD25 + regulatory T cells in spleen was detected by flow cytometry. Results In lungs of the asthmamodel group, there were intensive inflammatory cells infiltration around airway and blood vessels, goblet cell proliferation, epithelial desquamation, patchy airway occlusion by hyperviscous mucus, and hypertrophy of airway smooth muscle.Airway inflammation and airway remodeling were significantly relieved in the BMSCs transplantation group.There was no obvious inflammatory cells infiltration in the airway and airway remodeling both in the normal control group and the BMSCs control group. The number of CD4 + CD25 + regulatory T cells in spleensignificantly decreased in the asthma model group compared with the two control groups ( P lt; 0. 05) , and significantly increased in the BMSCs transplantation group compared with the asthma model group ( P lt;0. 05) . There was no significant difference in the number of CD4 + CD25 + regulatory T cells in spleen betweenthe control groups and the BMSCs transplantation group. Conclusion BMSCs engraftment can up-regulate CD4 + CD25 + regulatory T cells and relieve airway inflammation and airway remodeling in asthmatic mice.
Objective To improve our recognition of ground-glass opacity (GGO) through analyzing the imaging and pathological features of patients with focal GGO lung nodule. Methods Thirty patients with focal GGO nodule were assigned into a preinvasive lesion group, a minimally invasive adenocarcinoma (MIA) group, and an invasive adenocarcinoma (IAC) group. The imaging features were retrospectively analyzed and pathological features by histological Masson staining, collagen Ⅳ staining and Vitoria blue staining were also compared among three groups. Furthermore, the relationship between pathology and imaging characteristics was studied too. Results Among 30 patients with focal GGO nodule, preinvasive lesions, MIA and IAC respectively occurred in 13, 3 and 14 cases. Size of nodules and solid portion were highest in the IAC group, middle in the MIA group, and lowest in the preinvasive lesion group. Similarly, signs of lobulation, spiculation and air bronchogram were seen mostly in the IAC group, and least in preinvasive lesion group. The spatial relationship between GGO nodules and supplying blood vessels was analyzed, and Type Ⅲ was more commonly seen in the IAC group with comparison to type Ⅱ more likely seen in the preinvasive lesion group. Moreover, collagen Ⅳ and Vitoria blue staining indicated that reticular fibers and collagenous fibers lessened around tumor tissue in the IAC group, whereas collagenous fibers proliferation and fibrous scar were shown by Masson staining in the IAC group. In CT-pathologic comparison, type Ⅲ supplying blood vessels were mostly seen in the IAC patients with obvious fibrous scar. Conclusions Persistent focal GGO nodules with larger size and higher percent of solid component are signs of malignancy. In tumor progression process, tumor cells break the reticular fibers and collagenous fibers in alveolar wall, then stimulate fibroblast hyperplasia and secrete collagenous fibers, thereby develop the central fibrous scar in tumor tissue, which might be the pathologic foundation of vascular bundle sign.