HSP90在脓毒症小鼠中的表达及血必净对其表达的影响

The expression of HSP90 in septic mice and the intervention of Xuebijing injection

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。

To investigate the expression of heat shock pretein90 （HSP90） in septic mice, and the effect of Xuebijing intervention and treatment on the expression of HSP90. The biological significance of HSP90 factor was discussed in sepsis. The septic mice model was established by intravenous injection of lipopolysaccharides （LPS）. The mice in the intervention group were injected with Xuebijing of 10 mL/kg twice a day, while the mice in the control group were treated with the same dosage of saline solution. Real time quantitative PCR was used to analyze the relative gene expression of HSP90 in heart and kidney tissues. The level of serum creatinine （Cr）, blood urea nitrogen （BUN）, HSP90 were detected by ELISA kit. Finally, H. E. staining was used to observe the changes of pathological structures of heart and kidney tissues in two groups. The results showed that the serum levels of Cr and BUN increased,mad the highest level was at 18h after mice tail vein injection of LPS. The serum BUN and Cr levels of the two groups of mice were significantly different after injection of LPS 8 hours （ P 〈 0.05 ）. In the heart tissues of septic mice, mRNA and protein levels of HSP90 were significantly up - regulated. The whole tendency was increased and then decreased. The mRNA and protein levels of HSP90 in the control group were higher than that in Xuebijing intervention group （ P 〈 0.05, P 〈 0.01 ）, and there was a similar tendency of HsPgo between heart and kidney tissues. In the sepsis model group,inflammatory reaction obviously occurred in myocardial tissues and renal tissues, the pathological structure was abnormal. While the myocardial and kidney tissues maintained a regular and clear pathological structure in intervention group, there were no obvious pathological changes. The results showed that the mRNA and protein expressions of HSP90 in the heart and kidney tissues of septic mice were increased with the occurrence and development of sepsis in different degrees, it was found to be significantly higher in the first 2 hours after modeling,which suggested that HSP90 was involved in the occurrence and the development course of sepsis. It was concluded that HSP90 could be classified as one of the early clinical detective and diagnostic indicators of sepsis. Besides,Xuebijing intervention played an important role in regulating HSP90 factor,and could effectively improve the disease symptoms of sepsis.