| Objective To verify whether isosteviol can control the polarization of macrophages by affecting the expression level of mi R-146 a to achieve the effect of preventing or treating intestinal ischemia-reperfusion injury,and combining isosteviol and Treg cells to observe whether them can be further enhanced Macrophage anti-inflammatory effect,reduce intestinal ischemia-reperfusion injury more effectively.Methods First divide the mice into 5 groups,namely sham operation group,ischemia-reperfusion injury(I/R)group,low-dose administration group,middle-dose administration group and high-dose administration group,and then establish the mouse intestinal Ischemia-reperfusion injury model,and intervention with different doses of isosteviol to observe whether it can reduce intestinal ischemia-reperfusion injury and determine the best suitability of isosteviol dose.The Treg cells whose mi R-146 a level was changed by mi R-146 a agomir and antagomir were infused into mice.Then divide the mice into 5 groups,namely sham operation group,I/R group,isosteviol administration group,treg with mi RNA-146 a up-regulation combined with isosteviol administration group,treg with mi RNA-146 a down-regulation combined with isosteviol administration group,create intestinal ischemia-reperfusion injury model again and treat the mice in each group with the optimal dose of isosteviol to derive the mechanism of its effect on intestinal ischemia-reperfusion injury.Results The rate of apoptotic cells in I/R group was significantly higher than that in the sham operation group,the amount of Notch1 and NF-κB p65 and the secretion of inflammatory factors TNF-α and IL-1β were significantly increased(P<0.01).Compared with the I/R group,the apoptosis and M1 macrophage infiltration in three isosteviol groups was reduced,while M2 macrophage infiltration was increased.And the inflammatory factors TNF-α and IL-1β secretion were decreased(P<0.05).The secretion of anti-inflammatory factors TGF-β1 and IL-10 was increased(P<0.05),while the amount of Notch1 and NF-κB p65 and its m RNA expression were significantly reduced,and the m RNA level of mi R-146 a was significantly increased(P <0.05).Among them,the anti-inflammatory effect of the middle and high dose administration group is the same and stronger than that of the low dose administration group.In mi R-146 a up-regulating combined with isosteviol group,the infiltration of M1 macrophage was significantly lower than that in the single administration group,and the secretion of inflammatory factors TNF-α and IL-1β was significantly reduced(P<0.05),the secretion of anti-inflammatory factors TGF-β1 and IL-10 increased significantly(P<0.05).Similarly,as mi R-146 a further increased,Notch1 and NF-κB p65 expressions further decreased.While in mi R-146 a down-regulating combined with the isosteviol group,the secretion of inflammatory factors TNF-α and IL-1β was significantly increased(P<0.05).Conclusions The first phase of the experiment mainly explored the effect of isosteviol on macrophages,and obtained the best dose of isosteviol to protect the intestinal tissue of mice from ischemia-reperfusion injury.By comparing the difference in macrophage differentiation between the administration group and the I / R group,it can be concluded that isosteviol may protect the intestine by promoting the differentiation of macrophages into M2,increasing the secretion of anti-inflammatory factors and reducing the production of inflammatory factors.Further study of its mechanism,it can be found that isosteviol can inhibit the Notch1 signaling pathway by increasing the expression of mi R-146 a,reduce the activation of NF-κB p65 and reduce the differentiation of macrophages into M1 macrophages.The second stage demonstrated isosteviol may play a role by affecting the expression level of mi R-146 a in Treg cells. |
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