【摘要】：為了研究熱休克蛋白90(HSP90)因子在膿毒癥小鼠中的表達(dá)水平,并研究血必凈干預(yù)、治療對(duì)HSP90表達(dá)的影響,探討HSP90因子在膿毒癥病程中的生物學(xué)意義,試驗(yàn)采用尾靜脈注射脂多糖(LPS)建立膿毒癥小鼠模型,血必凈干預(yù)組小鼠腹腔注射10 m L/kg劑量血必凈,每日2次,而模型組小鼠予以等量生理鹽水處理,采用實(shí)時(shí)定量PCR(Real time PCR)方法檢測(cè)兩組小鼠心臟和腎臟組織中HSP90 mRNA的相對(duì)表達(dá)量;采用酶聯(lián)免疫吸附試驗(yàn)(ELISA)檢測(cè)兩組小鼠血清肌酐(Cr)、血尿素氮(BUN)水平及心臟、腎臟組織液中HSP90因子表達(dá)水平;采用H.E.染色觀察兩組小鼠心臟、腎臟組織病理結(jié)構(gòu)。結(jié)果表明:小鼠尾靜脈注射LPS后,血清Cr和BUN水平出現(xiàn)持續(xù)性增高,于18小時(shí)達(dá)到最高水平,兩組小鼠血清BUN、Cr水平最早于注射LPS造模8 h后出現(xiàn)顯著差異(P0.05);在膿毒癥小鼠心臟組織中,HSP90 mRNA及蛋白水平均出現(xiàn)明顯上調(diào),整體呈先上升后下降的趨勢(shì),模型組HSP90 mRNA及蛋白總體水平高于血必凈干預(yù)組(P0.05,P0.01),HSP90 mRNA及蛋白在膿毒癥小鼠腎臟組織中的表達(dá)水平及變化趨勢(shì)與其在心臟組織中相似;膿毒癥小鼠心、腎組織發(fā)生明顯炎性反應(yīng),可見(jiàn)組織結(jié)構(gòu)異常,而干預(yù)組小鼠保持較為完整的心臟、腎臟組織結(jié)構(gòu),無(wú)顯著病理學(xué)變化。說(shuō)明膿毒癥小鼠心臟、腎臟組織中的HSP90基因和蛋白的表達(dá)隨膿毒癥發(fā)生、發(fā)展表現(xiàn)不同程度的上升,最早于造模后2小時(shí)出現(xiàn)顯著增高,表明HSP90因子參與膿毒癥發(fā)生、發(fā)展病程,提示可將HSP90列為膿毒癥臨床早期檢測(cè)、診斷指標(biāo)之一;血必凈干預(yù)對(duì)HSP90因子具有調(diào)節(jié)作用,能夠有效改善膿毒癥癥狀。
[Abstract]:In order to study the expression of heat shock protein 90 (HSP90) factor in sepsis mice, and to study the effect of Xuebijing intervention on the expression of HSP90, and to explore the biological significance of HSP90 factor in the course of sepsis. The model of sepsis mice was established by intravenous injection of lipopolysaccharide (LPS). The mice in Xuebijing intervention group were intraperitoneally injected with 10 m L/kg Xuebijing twice a day, while the mice in model group were treated with the same amount of saline. The relative expression of HSP90 mRNA in heart and kidney of the two groups was detected by real-time quantitative PCR (Real time PCR). Enzyme linked immunosorbent assay (ELISA) was used to detect the level of serum creatinine (Cr), blood urea nitrogen (BUN) and the expression of HSP90 factor in heart and kidney tissue fluid of the two groups. The pathological structure of heart and kidney in the two groups was observed by staining. The results showed that the levels of serum Cr and BUN increased continuously after intravenous injection of LPS in mice, and reached the highest level at 18 hours. The levels of serum BUN,Cr in the two groups were significantly different from those 8 hours after injection of LPS (P 0.05). In the heart tissue of sepsis mice, the levels of HSP90 mRNA and protein were significantly up-regulated and increased at first and then decreased. The overall levels of HSP90 mRNA and protein in the model group were higher than those in the Xuebijing intervention group (P0.05, P01). The expression level and change trend of HSP90 mRNA and protein in kidney tissue of sepsis mice were similar to those in heart tissue. The heart and kidney tissue of sepsis mice had obvious inflammatory reaction and abnormal tissue structure, while the intervention group maintained a more complete heart and kidney tissue structure, and there was no significant pathological change. The results showed that the expression of HSP90 gene and protein in heart and kidney of sepsis mice increased in varying degrees with the occurrence of sepsis, and increased significantly as early as 2 hours after modeling, indicating that HSP90 factor was involved in the occurrence of sepsis. The development of the course of disease suggests that HSP90 can be listed as one of the early clinical detection and diagnostic indexes of sepsis. Xuebijing intervention can regulate HSP90 factor and effectively improve sepsis symptoms.
【作者單位】： 南京醫(yī)科大學(xué)康達(dá)學(xué)院基礎(chǔ)醫(yī)學(xué)部;南通大學(xué)實(shí)驗(yàn)動(dòng)物中心;
【基金】：江蘇省高校自然科學(xué)研究面上項(xiàng)目(16KJD180005) 南京醫(yī)科大學(xué)康達(dá)學(xué)院科研發(fā)展基金項(xiàng)目(KD2015KYJJYB001)
【分類(lèi)號(hào)】：R459.7

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