PKCβ抑制剂通过影响巨噬细胞亚型的改变对肾缺血再灌注损伤起保护作用

发布时间：2018-08-29 18:56

【摘要】：目的:采用肾缺血再灌注损伤为模型,经口给予蛋白激酶Cβ(PKCβ)抑制剂干预,观察PKCβ抑制剂对肾缺血再灌注损伤中炎性介质及巨噬细胞等的影响,探讨其对肾IRI的保护机制。方法:将购买的大鼠随机分为三组,分别为肾缺血再灌注损伤模型组(A组),PKCβ抑制剂干预组(B组)和假手术组(C组)。A组予去右肾,左肾按常规方式即肾蒂钳夹缺血60分钟后恢复灌注24小时;B组予术前经口腔给药PKCβ抑制剂干预;C组仅行开腹、60分钟后关腹手术。术后24小时取下腔静脉血及肾脏标本进行实验研究:1.全自动生化仪检测各组血清肌酐值及尿素情况;2.PAS染色观察病理组织损伤严重程度;3.免疫组织化学法检测肾损伤分子-1、肾乳头状抗原-1及诱生型一氧化氮合酶-1、精氨酸酶-1的表达;结果:1.全自动生化仪检测血清肌酐、尿素结果示:尿素值A、B组均较C组增高(P0.01),且A组水平高于B组(P0.01);同样肌酐值A、B组也均高于C组(P0.01),但A组与B组比较无明显差异(P0.05);2.肾组织PAS染色结果:A组肾间质水肿、炎性细胞浸润,肾小管严重破坏、结构紊乱、管腔扩张,可见小管刷状缘丢失、管状上皮细胞扁平、脱落,并有大量管型存在;B组:相较A组,小管结构较清晰,肾间质水肿及炎性细胞浸润减轻,但仍可见部分小管结构破坏,偶见管型;C组:小管结构清晰、完整,未见管型及明显间质水肿,但组织间隙中可见少量炎性细胞浸润。3.免疫组化检测:KIM-1结果显示在C组少量表达,A组、B组表达均较C组明显升高(P0.01),而A组又显著强于B组(P0.01);RPA-1示A组在肾乳头小管强阳性表达,B组表达较A组明显减轻(P0.01),C组微量或基本未见表达。同样iNOS的表达也显示A组明显强于B、C组(P0.01),C组表达甚少,B组强于C组(P0.01);而在Arg-1的表达上B组较A、C组均强(P0.01),而A组又高于C组(P0.01)。结论:PKCβ抑制剂可以通过影响巨噬细胞亚型的变化而产生保护肾缺血再灌注损伤的作用。
[Abstract]:Objective: to observe the effects of protein kinase C 尾 (PKC 尾) inhibitor on inflammatory mediators and macrophages in renal ischemia-reperfusion injury and explore the protective mechanism of PKC 尾 on renal IRI. Methods: the rats were randomly divided into three groups: model group (group A), PKC 尾 inhibitor intervention group (group B) and sham operation group (group C). The left kidney was treated with oral administration of PKC 尾 inhibitor for 60 minutes after ischemia for 60 minutes and then restored to perfusion for 24 hours. Group C was treated with open abdominal surgery for 60 minutes and closed abdominal operation after 60 minutes. Blood samples of inferior vena cava and kidney were collected 24 hours after operation. The levels of serum creatinine and urea were detected by automatic biochemical instrument. 2. The severity of pathological tissue injury was observed by pas staining. The expression of renal injury molecule -1, papillary antigen 1 and inducible nitric oxide synthase 1 and arginase 1 were detected by immunohistochemistry. Serum creatinine was detected by automatic biochemical instrument. Urea value in group A was higher than that in group C (P0.01), and the level of creatinine in group A was higher than that in group B (P0.01). Similarly, creatinine level in group A was higher than that in group C (P0.01), but there was no significant difference between group A and group B (P0.05). Renal tissue PAS staining showed interstitial edema, inflammatory cell infiltration, serious destruction of renal tubules, structural disorder, dilatation of lumen, loss of brush edge of tubules, flattening and shedding of tubular epithelial cells in group A. There were a large number of tubules in group B: compared with group A, the tubule structure was clearer, renal interstitial edema and inflammatory cell infiltration were alleviated, but some tubules were still damaged. Occasionally, tubule structure was clear and complete in group C. No tubular and interstitial edema was found, but a small number of inflammatory cells infiltrated in the interstitial space. 3. Immunohistochemical analysis showed that the expression of KIM-1 in group A was significantly higher than that in group C (P0.01), while the expression of RPA-1 in group A was significantly stronger than that in group B (P0.01). The positive expression of KIM-1 in group A was significantly lower than that in group A (P0.01). Group A showed little or no expression. The expression of iNOS in group A was significantly higher than that in group C (P0.01), but the expression of Arg-1 in group B was stronger than that in group C (P0.01), and the expression of Arg-1 in group A was higher than that in group C (P0.01). Conclusion the effect of PKC 尾 inhibitor on renal ischemia-reperfusion injury may be induced by affecting the changes of macrophage subtypes.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R692

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