干细胞移植对实验性纤维化大鼠肺形态功能状态的恢复

IF 0.4 Q4 AGRICULTURE, MULTIDISCIPLINARY Agricultural Science and Practice Pub Date : 2023-08-02 DOI:10.15407/agrisp10.01.012
A. Mazurkevych, Y. Surtaieva
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In the first experimental group, allogeneic MSC were administered intrathoracically. In the second experimental group, allogeneic MSC were administered intravenously. The third experimental group received conventional treatment using medication, while the fourth experimental group (control) received placebo intrathoracically, without any active agents. Additionally, a separate fifth experimental group comprised intact (healthy) animals. The stem cells used in the experiments were obtained from the bone marrow samples of young, clinically healthy donor rats’ tibia, humerus, or femur (Mazurkevych et al, 2014). Throughout the 45-day experiment, the animals in all experimental groups were monitored using clinical examination indices. At the end of the testing period, the animals were eutha- nized, and blood, bronchoalveolar lavage, and lung tissue samples were collected for laboratory analyses. The analysis of bronchoalveolar lavage involved microscopic examination of the collected fluid to determine cell composition, while hematological analysis encompassed the quantification of erythrocyte and leukocyte counts. Histological examination of lung tissue samples involved microscopic analysis of the lung tissue histostructure. Results. Following the implementation of the proposed treatment methods, it was observed on day 45 of the experiment that transplanted allogeneic MSC facilitated an increased activity in the restoration of pathologically altered lung parenchyma. The effectiveness of this process varied depending on the method of MSC application. Notably, animals in the first experimental group exhibited the absence of symptoms such as coughing and hypoxia by day 45. Furthermore, a significant decrease was observed in the indices of erythrocyte count to 7.18 ± 0.05 t/l (р < 0.001) and total leukocyte count to 10.6 ± 0.92 g/l (р < 0.05). Histological analysis revealed a complete restoration of lung tissue structure, reaching 90 % when compared to the control group. In the second experimental group, a significant decrease was noted in the total leukocyte count to 11.32 ± 0.48 g/l (р < 0.01) and erythrocyte count to 6.87 ± 0.18 t/l (р < 0.001). Histologically, the regenerative processes exhibited lower activity, reaching 70–80 % compared to the animals in the first group. Fibrous areas were observed but of smaller size than those in the conventional treatment group. In the third experimental group, despite the normalization of laboratory testing indices, namely a decrease in erythrocyte count to 7.62 ± 0.11 t/l (р < 0.001) and leukocyte count to 9.46 ± 0.54 g/l (р < 0.001), occasional fibrosis areas and thicker alveolar walls were still present in the histological structure of the lungs. The fibrosis areas decreased by only 20–30 %. Conclusions. 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引用次数: 0

摘要

的目标。目的探讨骨髓异体间充质干细胞(MSC)对实验性肺纤维化大鼠肺组织损伤的修复作用。方法。本研究采用雌性wistar大鼠。在45天的预备期,经胸注射盐酸博来霉素实验性地诱导肺纤维化。在这段时间结束时,所有动物都表现出临床表现的肺纤维化症状。为了评估肺纤维化引起的机体变化,随机收集了5只患病动物的血液样本、支气管肺泡灌洗液样本和肺组织。剩下的动物被分成4个实验组,每组5只。在第一个实验组,同种异体间充质干细胞给予胸腔内注射。在第二个实验组,异体间充质干细胞静脉注射。第三实验组采用常规药物治疗,而第四实验组(对照组)采用胸内注射安慰剂,不使用任何活性药物。此外,第五个实验组由完整(健康)的动物组成。实验中使用的干细胞是从年轻、临床健康的供体大鼠的胫骨、肱骨或股骨的骨髓样本中获得的(Mazurkevych et al ., 2014)。在45 d的实验期间,采用临床检查指标对各组动物进行监测。实验结束后,对动物进行安乐死,采集血液、支气管肺泡灌洗液和肺组织样本进行实验室分析。支气管肺泡灌洗分析包括显微镜检查收集的液体以确定细胞组成,而血液学分析包括红细胞和白细胞计数的定量。肺组织样本的组织学检查包括肺组织组织结构的显微分析。结果。在实施上述治疗方法后,在实验第45天观察到,移植的同种异体间充质干细胞促进了病理改变的肺实质恢复的活性增加。这一过程的有效性取决于MSC应用的方法。值得注意的是,第一个实验组的动物在第45天没有出现咳嗽和缺氧等症状。红细胞计数降至7.18±0.05 t/l(< 0.001),白细胞总数降至10.6±0.92 g/l(< 0.05)。组织学分析显示,与对照组相比,肺组织结构完全恢复,达到90%。第二组白细胞总数明显下降至11.32±0.48 g/l(< 0.01),红细胞总数明显下降至6.87±0.18 t/l(< 0.001)。组织学上,再生过程表现出较低的活性,与第一组动物相比,达到70 - 80%。观察到纤维区,但比常规治疗组小。在第三组,尽管实验室检测指标正常化,即红细胞计数降至7.62±0.11 g/l(< 0.001),白细胞计数降至9.46±0.54 g/l(< 0.001),但肺组织结构中仍存在偶尔的纤维化区和较厚的肺泡壁。纤维化面积仅减少20 - 30%。结论。我们的研究结果表明,与常规治疗相比,间充质干细胞的应用是一种更有效的细胞治疗方法,可用于实验性诱导肺纤维化大鼠病理改变的肺组织的修复。在45天的实验中,传统药物治疗使临床指标和实验室检查恢复正常,但未导致受损肺组织结构的完全恢复。这些结果强调了同种异体间充质干细胞在肺纤维化治疗中的优势,表明其具有进一步研究和临床应用的潜力。
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Restoration of the morphofunctional state of rats lungs with experimental fibrosis through transplanted stem cells
Aim. To investigate the therapeutic effect of allogeneic mesenchymal stem cells (MSC) derived from bone marrow on the restoration experimentally damaged lung tissue in rats with induced pulmonary fibrosis. Methods. Female Wistar rats were utilized in the study. Pulmonary fibrosis was induced experimentally in the animals by administering bleomycin hydrochloride via transthoracic injection during a 45-day preparatory period. At the end of this period, all animals exhibited clinically manifested symptoms of pulmonary fibrosis. To assess the changes in the organism due to pulmonary fibrosis, blood samples, bronchoalveolar lavage samples, and lung tissue were randomly collected from five affected animals. The remaining animals were divided into four experimental groups, each consisting of five animals. In the first experimental group, allogeneic MSC were administered intrathoracically. In the second experimental group, allogeneic MSC were administered intravenously. The third experimental group received conventional treatment using medication, while the fourth experimental group (control) received placebo intrathoracically, without any active agents. Additionally, a separate fifth experimental group comprised intact (healthy) animals. The stem cells used in the experiments were obtained from the bone marrow samples of young, clinically healthy donor rats’ tibia, humerus, or femur (Mazurkevych et al, 2014). Throughout the 45-day experiment, the animals in all experimental groups were monitored using clinical examination indices. At the end of the testing period, the animals were eutha- nized, and blood, bronchoalveolar lavage, and lung tissue samples were collected for laboratory analyses. The analysis of bronchoalveolar lavage involved microscopic examination of the collected fluid to determine cell composition, while hematological analysis encompassed the quantification of erythrocyte and leukocyte counts. Histological examination of lung tissue samples involved microscopic analysis of the lung tissue histostructure. Results. Following the implementation of the proposed treatment methods, it was observed on day 45 of the experiment that transplanted allogeneic MSC facilitated an increased activity in the restoration of pathologically altered lung parenchyma. The effectiveness of this process varied depending on the method of MSC application. Notably, animals in the first experimental group exhibited the absence of symptoms such as coughing and hypoxia by day 45. Furthermore, a significant decrease was observed in the indices of erythrocyte count to 7.18 ± 0.05 t/l (р < 0.001) and total leukocyte count to 10.6 ± 0.92 g/l (р < 0.05). Histological analysis revealed a complete restoration of lung tissue structure, reaching 90 % when compared to the control group. In the second experimental group, a significant decrease was noted in the total leukocyte count to 11.32 ± 0.48 g/l (р < 0.01) and erythrocyte count to 6.87 ± 0.18 t/l (р < 0.001). Histologically, the regenerative processes exhibited lower activity, reaching 70–80 % compared to the animals in the first group. Fibrous areas were observed but of smaller size than those in the conventional treatment group. In the third experimental group, despite the normalization of laboratory testing indices, namely a decrease in erythrocyte count to 7.62 ± 0.11 t/l (р < 0.001) and leukocyte count to 9.46 ± 0.54 g/l (р < 0.001), occasional fibrosis areas and thicker alveolar walls were still present in the histological structure of the lungs. The fibrosis areas decreased by only 20–30 %. Conclusions. Our findings demonstrate that the application of mesenchymal stem cells is a more effective approach to cell therapy for the restoration of pathologically altered lung tissue in rats with experimentally induced pulmonary fibrosis compared to conventional treatment. Traditional medication-based treatment over the 45-day experiment led to the normalization of clinical indices and laboratory tests but did not result in complete restoration of the damaged lung tissue structure. These results underscore the advantages of employing allogeneic mesenchymal stem cells in the therapy of pulmonary fi brosis, indicating their potential for further investigation and clinical application.
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Agricultural Science and Practice
Agricultural Science and Practice AGRICULTURE, MULTIDISCIPLINARY-
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