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为了研究热休克蛋白90(HSP90)因子在脓毒症小鼠中的表达水平,并研究血必净干预、治疗对HSP90表达的影响,探讨HSP90因子在脓毒症病程中的生物学意义,试验采用尾静脉注射脂多糖(LPS)建立脓毒症小鼠模型,血必净干预组小鼠腹腔注射10 m L/kg剂量血必净,每日2次,而模型组小鼠予以等量生理盐水处理,采用实时定量PCR(Real time PCR)方法检测两组小鼠心脏和肾脏组织中HSP90 mRNA的相对表达量;采用酶联免疫吸附试验(ELISA)检测两组小鼠血清肌酐(Cr)、血尿素氮(BUN)水平及心脏、肾脏组织液中HSP90因子表达水平;采用H.E.染色观察两组小鼠心脏、肾脏组织病理结构。结果表明:小鼠尾静脉注射LPS后,血清Cr和BUN水平出现持续性增高,于18小时达到最高水平,两组小鼠血清BUN、Cr水平最早于注射LPS造模8 h后出现显著差异(P<0.05);在脓毒症小鼠心脏组织中,HSP90 mRNA及蛋白水平均出现明显上调,整体呈先上升后下降的趋势,模型组HSP90 mRNA及蛋白总体水平高于血必净干预组(P<0.05,P<0.01),HSP90 mRNA及蛋白在脓毒症小鼠肾脏组织中的表达水平及变化趋势与其在心脏组织中相似;脓毒症小鼠心、肾组织发生明显炎性反应,可见组织结构异常,而干预组小鼠保持较为完整的心脏、肾脏组织结构,无显著病理学变化。说明脓毒症小鼠心脏、肾脏组织中的HSP90基因和蛋白的表达随脓毒症发生、发展表现不同程度的上升,最早于造模后2小时出现显著增高,表明HSP90因子参与脓毒症发生、发展病程,提示可将HSP90列为脓毒症临床早期检测、诊断指标之一;血必净干预对HSP90因子具有调节作用,能够有效改善脓毒症症状。
In order to study the expression of heat shock protein 90 (HSP90) in septic mice and the effect of Xuebijing intervention on the expression of HSP90, to explore the biological significance of HSP90 in the course of sepsis, the experiment The mouse model of sepsis was established by injecting lipopolysaccharide (LPS) via tail vein. The mice in Xuebijing intervention group were injected with Xuebijing (10 ml / kg) twice a day by intraperitoneal injection. The mice in model group were given the same amount of physiology The expression of HSP90 mRNA in the heart and kidney of the two groups was detected by real time PCR. The serum creatinine (Cr) was measured by enzyme linked immunosorbent assay (ELISA) Blood urea nitrogen (BUN) levels and heart and kidney tissue fluid HSP90 factor expression levels; using HE staining of the two groups of mice heart and kidney histopathology. The results showed that the levels of serum Cr and BUN increased continuously after the tail vein injection of LPS, reaching the highest level at 18 hours. The serum levels of BUN and Cr in the two groups were significantly different at 8 h after LPS injection P <0.05). HSP90 mRNA and protein levels were significantly up-regulated in heart tissue of septic mice at first increasing and then decreasing. The levels of HSP90 mRNA and protein in the model group were higher than those in the Xuebijing intervention group P <0.05, P <0.01). HSP90 mRNA and protein expression in renal tissues of septic mice was similar to that of HSP90 mRNA and protein in cardiac tissue. Serum and protein expressions of HSP90 mRNA and protein in sepsis mice were significantly increased Visible organizational structure abnormalities, while the intervention group mice to maintain a more complete heart, kidney tissue structure, no significant pathological changes. The results showed that the HSP90 gene and protein expression in sepsis mice heart and kidney tissue with the sepsis, the development of varying degrees of increase, as early as 2 hours after modeling showed a significant increase, indicating that HSP90 factor involved in the occurrence of sepsis , Suggesting that HSP90 may be one of the early detection and diagnostic indicators of sepsis. The intervention of Xuebijing could regulate HSP90 and improve the symptoms of sepsis.