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目的急性肝损伤过程中,肝脏内存在来源不同的两类巨噬细胞即肝脏固有巨噬细胞和单核细胞源性巨噬细胞。该研究明确这两类巨噬细胞在急性肝损伤过程中的作用。方法采用腹腔注射四氯化碳(CCl4)构建急性肝损伤小鼠模型,通过流式细胞术分析注射CCl4后24、48、72 h时小鼠肝内巨噬细胞表面抗原(CD45,F4/80,Ly6C和CD11b)的表达情况,分选出肝脏固有巨噬细胞(CD45+F4/80hi)和单核细胞源性巨噬细胞(CD45+F4/80lo),并应用qRT-PCR检测两群巨噬细胞中细胞因子的表达情况。结果与正常对照相比,注射CCl4后24、48、72 h时小鼠肝内总F4/80+细胞数显著增加,尤其是72 h时;24 h CD45+F4/80lo细胞数显著增加。急性肝损伤过程中,与正常对照相比,F4/80hi巨噬细胞和F4/80lo巨噬细胞中MCP-1、TNF-α、TGF-β1、基质金属蛋白酶(MMP)-12和MMP-13mRNA水平均显著升高,IL-12β和IL-6 mRNA水平只在F4/80hi巨噬细胞中显著升高,MMP-9 mRNA水平只在F4/80lo巨噬细胞中显著升高。与F4/80lo巨噬细胞相比,F4/80hi巨噬细胞中MCP-1和MMP-12 mRNA水平显著升高,TNF-αmRNA水平显著降低。结论急性肝损伤时,肝内固有巨噬细胞和单核细胞源性巨噬细胞均表达炎性细胞因子,促进炎症反应,导致肝损伤;肝内固有巨噬细胞募集炎性单核细胞的能力和表达炎性细胞因子(IL-12β,IL-6)、MMP-12的能力强于单核细胞源性巨噬细胞;单核细胞源性巨噬细胞表达炎性细胞因子(TNF-α)和MMP-9的能力强于固有巨噬细胞。
Aim In the process of acute liver injury, there are two types of macrophages from different sources, liver intrinsic macrophages and monocyte-derived macrophages. The study identified the role of these two types of macrophages in acute liver injury. Methods Acute hepatic injury model was established by intraperitoneal injection of carbon tetrachloride (CCl4). The expression of macrophages surface antigens (CD45, F4 / 80) in mice after 24, 48 and 72 h injection of CCl4 were analyzed by flow cytometry , Ly6C and CD11b) were detected by immunohistochemistry. Liver intrinsic macrophages (CD45 + F4 / 80hi) and monocyte-derived macrophages (CD45 + F4 / 80lo) were isolated and identified by qRT- Cytokine expression in macrophages. Results Compared with the normal control group, the total number of F4 / 80 + cells in the liver increased significantly at 24 h, 48 h and 72 h after CCl4 injection, especially at 72 h. The number of CD45 + F4 / 80 Lo cells increased significantly at 24 h. MCP-1, TNF-α, TGF-β1, MMP-12 and MMP-13 mRNA levels in F4 / 80hi macrophages and F4 / 80lo macrophages during acute liver injury The levels of IL-12β and IL-6 mRNA were significantly increased only in F4 / 80hi macrophages and the levels of MMP-9 mRNA were only significantly increased in F4 / 80lo macrophages. Compared with F4 / 80lo macrophages, MCP-1 and MMP-12 mRNA levels in F4 / 80hi macrophages were significantly increased and TNF-αmRNA levels were significantly reduced. Conclusions In acute liver injury, both macrophages and monocyte-derived macrophages in the liver express inflammatory cytokines and promote inflammatory reaction, leading to liver injury. The ability of intrinsic macrophages to raise inflammatory monocytes in the liver And the expression of inflammatory cytokines (IL-12β, IL-6), MMP-12 stronger than monocyte-derived macrophages; monocyte-derived macrophages expressed inflammatory cytokines (TNF-α) And MMP-9 are stronger than innate macrophages.