Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (13): 1969-1975.doi: 10.3969/j.issn.2095-4344.2195
Yuan Baohong, Zhao Weishan, Wang Ruotian, Guan Aoran, Wang Yu, Li Ruhong
Received:
2020-01-07
Revised:
2020-01-14
Accepted:
2020-04-27
Online:
2021-05-08
Published:
2020-12-28
Contact:
Li Ruhong, MD, Chief physician, Doctoral supervisor, First Department of General Surgery, Yan’an Hospital of Kunming City (Affiliated Yan’an Hospital of Kunming Medical University), Kunming 650051, Yunnan Province, China
About author:
Yuan Baohong, Master, Attending physician, First Department of General Surgery, Yan’an Hospital of Kunming City (Affiliated Yan’an Hospital of Kunming Medical University), Kunming 650051, Yunnan Province, China
Zhao Weishan, Associate chief physician, First Department of General Surgery, Yan’an Hospital of Kunming City (Affiliated Yan’an Hospital of Kunming Medical University), Kunming 650051, Yunnan Province, China
Supported by:
CLC Number:
Yuan Baohong, Zhao Weishan, Wang Ruotian, Guan Aoran, Wang Yu, Li Ruhong. Bone marrow mesenchymal stem cells from pigs inhibit inflammatory response induced by lipopolysaccharide and improve islet cell dysfunction in pigs[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(13): 1969-1975.
2.1 猪骨髓间充质干细胞改善脂多糖引起的胰岛细胞功能损伤 如图1所示,脂多糖处理猪胰岛细胞12 h后,细胞中白细胞介素6、白细胞介素1β、肿瘤坏死因子α及活性氧水平显著上调,超氧化物歧化酶及胰岛素水平显著下调(均P < 0.01);猪骨髓间充质干细胞与脂多糖处理12 h后的胰岛细胞共培养后,恢复了脂多糖对猪胰岛细胞的促炎和促氧化应激的作用(均P < 0.05)。 如图1D所示,Hoechst 33258染色后在荧光显微镜下可见正常对照组猪胰岛细胞被均匀染色,没有明显的凋亡荧光信号;脂多糖处理猪胰岛细胞12 h后,细胞凋亡荧光信号明显增强;猪骨髓间充质干细胞与脂多糖处理12 h后的胰岛细胞共培养后,恢复了脂多糖对猪胰岛细胞凋亡荧光信号的促进作用。 如图1E所示,Annexin V-FITC/PI检测猪胰岛细胞凋亡率,脂多糖处理猪胰岛细胞12 h后,猪胰岛细胞凋亡率显著增加(均P < 0.05);猪骨髓间充质干细胞与脂多糖处理12 h后的胰岛细胞共培养后,恢复了脂多糖对猪胰岛细胞凋亡的促进作用(均P < 0.05)。由此可知,猪骨髓间充质干细胞改善了脂多糖引起的胰岛细胞功能损伤。 "
2.5 猪骨髓间充质干细胞通过miR-299-5p/SIAH1分子轴抑制JNK/C-Jun信号通路改善脂多糖引起的猪胰岛细胞功能损伤 ELISA检测结果如图5所示,与脂多糖处理组相比,脂多糖和骨髓间充质干细胞共同处理组、脂多糖和骨髓间充质干细胞共同处理后同时敲降miR-299-5p和SIAH1组、SP600125干预脂多糖和骨髓间充质干细胞共同处理后敲降miR-299-5p组的胰岛细胞中白细胞介素6、白细胞介素1β、肿瘤坏死因子α及活性氧水平显著降低,超氧化物歧化酶表达及胰岛素水平显著增加(均P < 0.05);而脂多糖和骨髓间充质干细胞共同处理后敲降miR-299-5p组、脂多糖和骨髓间充质干细胞及Anisomycin共同处理组、SP600125干预脂多糖和骨髓间充质干细胞共同处理后同时敲降miR-299-5p和SIAH1组胰岛细胞中白细胞介素6、白细胞介素1β、肿瘤坏死因子α、活性氧、超氧化物歧化酶及胰岛素水平与脂多糖处理组无显著差异。 Hoechst 33258染色结果如图5D所示,脂多糖和骨髓间充质干细胞共同处理组、脂多糖和骨髓间充质干细胞共同处理后同时敲降miR-299-5p和SIAH1组、SP600125干预脂多糖和骨髓间充质干细胞共同处理后敲降miR-299-5p组的胰岛细胞被均匀染色,没有明显的凋亡荧光信号,而脂多糖和骨髓间充质干细胞共同处理后敲降miR-299-5p组、脂多糖和骨髓间充质干细胞及Anisomycin共同处理组、SP600125干预脂多糖和骨髓间充质干细胞共同处理后同时敲降miR-299-5p和SIAH1组的胰岛细胞中凋亡荧光信号明显增强。 Annexin V-FITC/PI 检测结果如图5E所示,与脂多糖处理组相比,脂多糖和骨髓间充质干细胞共同处理组、脂多糖和骨髓间充质干细胞共同处理后同时敲降miR-299-5p和SIAH1组、SP600125干预脂多糖和骨髓间充质干细胞共同处理后敲降miR-299-5p组的胰岛细胞凋亡率显著降低(均P < 0.05);而脂多糖和骨髓间充质干细胞共同处理后敲降miR-299-5p组、脂多糖和骨髓间充质干细胞及Anisomycin共同处理组、SP600125干预脂多糖和骨髓间充质干细胞共同处理后同时敲降miR-299-5p和SIAH1组的胰岛细胞凋亡率与脂多糖处理组无显著差异。由此可知,猪骨髓间充质干细胞通过调控 miR-299-5p/SIAH1分子轴抑制JNK/C-Jun信号通路,从而改善脂多糖引起的猪胰岛细胞功能损伤。 "
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