袁睿, 张宇, 刘时飞, 杨萌萌, 王陆, 赵妍, 李云, 邓子辉, 康红军. 间充质干细胞对热射病大鼠肝损伤的保护作用研究[J]. 解放军医学院学报, 2020, 41(12): 1211-1215. DOI: 10.3969/j.issn.2095-5227.2020.12.010
引用本文: 袁睿, 张宇, 刘时飞, 杨萌萌, 王陆, 赵妍, 李云, 邓子辉, 康红军. 间充质干细胞对热射病大鼠肝损伤的保护作用研究[J]. 解放军医学院学报, 2020, 41(12): 1211-1215. DOI: 10.3969/j.issn.2095-5227.2020.12.010
YUAN Rui, ZHANG Yu, LIU Shifei, YANG Mengmeng, WANG Lu, ZHAO Yan, LI Yun, DENG Zihui, KANG Hongjun. Protective effect of adipose-derived mesenchymal stem cells on liver injury in rats with heat stroke[J]. ACADEMIC JOURNAL OF CHINESE PLA MEDICAL SCHOOL, 2020, 41(12): 1211-1215. DOI: 10.3969/j.issn.2095-5227.2020.12.010
Citation: YUAN Rui, ZHANG Yu, LIU Shifei, YANG Mengmeng, WANG Lu, ZHAO Yan, LI Yun, DENG Zihui, KANG Hongjun. Protective effect of adipose-derived mesenchymal stem cells on liver injury in rats with heat stroke[J]. ACADEMIC JOURNAL OF CHINESE PLA MEDICAL SCHOOL, 2020, 41(12): 1211-1215. DOI: 10.3969/j.issn.2095-5227.2020.12.010

间充质干细胞对热射病大鼠肝损伤的保护作用研究

Protective effect of adipose-derived mesenchymal stem cells on liver injury in rats with heat stroke

  • 摘要:
      目的  探索脂肪来源间充质干细胞(mesenchymal stem cells,MSCs)对热射病(heat stroke,HS)大鼠肝损伤的保护作用。
      方法  从5只未成熟SD大鼠体内分离培养脂肪来源间充质干细胞,传代到P4备用。将30只SD大鼠随机分为3组:对照组6只,模型组(HS)12只,间充质干细胞治疗组(HS+MSCs)12只。对照组在25℃、40%湿度下室内饲养;HS组与HS+MSCs组分别置于40℃、45%湿度生化培养箱中,持续监测直肠温度,直到核心体温达42℃,大鼠出现肢体运动功能障碍,即认为HS大鼠模型构建成功。取出大鼠后置于室温,HS+MSCs组尾静脉注射悬浮于0.3 mL 0.9%氯化钠注射液(normalsaline,NS)中的MSCs(2×106),HS组给予0.3 mL NS尾静脉注射。两组大鼠分别于不同时间点(3 d、14 d)进行肝取材固定。采用HE染色方法观察肝的病理变化,利用Luminex平台检测大鼠肝组织中6种炎性因子的水平。
      结果  核心体温达到42℃后,大鼠造模成功。大鼠的肝HE染色显示炎症细胞浸润、肝细胞排列紊乱、点/片状出血、气球样变等。MSCs治疗的HS大鼠可见其肝组织病理改变较HS大鼠轻。促炎因子:HS组大鼠肝组织中白细胞介素(interleukin,IL)-1β、IL-6、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平均显著高于对照组(P<0.05),HS+MSCs大鼠仅IL-1β高于对照组(P<0.05),而IL-1β、IL-6、TNF-α水平在各时间点大多低于HS组大鼠(P<0.05)。抗炎因子:HS组和HS-MSCs组IL-10、IL-13水平均高于对照组(P<0.05),3 d时HS-MSCs组IL-10、IL-13显著高于HS组(P<0.05),14 d时差异无统计学意义(P>0.05);IL-4在各组间无统计学差异(P>0.05)。
      结论  MSCs治疗的HS大鼠促炎因子水平下降,抗炎因子水平上升,说明MSCs对HS大鼠肝有明确的保护作用。

     

    Abstract:
      Objective  To explore the protective effect of adipose derived mesenchymal stem cells (MSCs) on liver injury in rats with heat stroke.
      Methods  MSCs were isolated from 5 immature SD rats and subcultured to P4. Thirty SD rats were randomly divided into three groups: control group (n=6), model(HS) group (n=12) and MSCs (HS+MSCs) treatment group (n=12). The control group was reared in room temperature of 25℃ and humidity of 40%; the HS group and HS+MSCs group were placed in biochemical incubator at 40℃ and 45% humidity respectively, and the rectal temperature was continuously monitored until the core temperature reached 42℃, as the rats showed limb motor dysfunction, it suggested that the HS rat model was successfully constructed. Then the rats were removed and placed at room temperature. MSCs (2×106) suspended in 0.3 mL 0.9% sodium chloride solution (NS) were injected into the caudal vein of rats in the HS+MSCs group, and 0.3 mL NS was injected into the caudal vein of rats in the HS group. The liver samples were taken and fixed at different time points (3 d, 14 d). HE staining was used to observe the pathological changes of liver, and Luminex platform was used to detect the levels of six inflammatory factors in rats' liver tissues.
      Results  After the core temperature reached 42℃, the rat model was established successfully. HE staining results of the rat liver showed inflammatory cell infiltration, disordered arrangement of hepatocytes, spotty and patchy hemorrhage, balloon like degeneration, etc. The pathological changes of liver tissue in the HS rats treated with MSCs were milder than those in the HS rats. In terms of pro-inflammatory factors, the levels of IL-1 β, IL-6 and TNF - α in liver tissue of the HS group were significantly higher than those of the control group (P<0.05). Only IL-1 β of the HS MSCs rats was higher than that of the control group (P<0.05), while the levels of IL-1 β, IL-6 and TNF - α were mostly lower than those of the HS group at each time point (P<0.05). For anti-inflammatory factors, the levels of IL-10 and IL-13 in the HS group were higher than those in the HS MSCs group (P<0.05), and those in both of the above two groups were significantly higher compared to the control group (P<0.05) at day 3 (P<0.05), but there was no significant difference between them the three groups at day 14 (P>0.05); There was no significant difference in IL-4 between the three groups (P>0.05).
      Conclusion  HS rats treated with MSCs have lower level of pro-inflammatory factors and higher level of anti-inflammatory factors compared to the HS rats, which indicates that MSCs have a definitely protective effect on the liver of HS rats.

     

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