本文選題：膿毒癥 + 急性腎損傷�。� 參考：《山東大學(xué)》2016年碩士論文

【摘要】：膿毒癥是一種由感染引起的,以全身性炎癥反應(yīng)為特征的復(fù)雜的臨床綜合征。膿毒癥通常導(dǎo)致廣泛的組織損傷和多器官功能紊亂。急性腎損傷(acute kidney injury, AKI)是膿毒癥最常見的并發(fā)癥之一,不僅增加了疾病的復(fù)雜性和護(hù)理費(fèi)用,也是引起膿毒癥相關(guān)死亡的高風(fēng)險(xiǎn)因素。膿毒癥誘導(dǎo)的AKI的病理學(xué)機(jī)制十分復(fù)雜,涉及腎臟內(nèi)血流動(dòng)力學(xué)變化、內(nèi)皮細(xì)胞失調(diào)、腎實(shí)質(zhì)組織的炎性細(xì)胞浸潤(rùn)、腎小球血栓和壞死細(xì)胞與碎片引起的腎小管障礙等多種因素。Progranulin (PGRN)是一種新型的分泌生長(zhǎng)因子,在快速增殖的上皮細(xì)胞和特定類型的神經(jīng)細(xì)胞中高表達(dá),多種其他組織和細(xì)胞類型中也存在PGRN表達(dá),包括脂肪組織、造血細(xì)胞、軟骨細(xì)胞及免疫細(xì)胞等。PGRN具有多種生理功能,不僅維持和調(diào)控正常組織的發(fā)生發(fā)展、增殖、再生及宿主防御,也參與多種類型的疾病過(guò)程,包括自身免疫性疾病、癌癥和神經(jīng)退行性疾病等。近年來(lái),PGRN在炎癥方面的作用受到廣泛關(guān)注,另有研究報(bào)道,PGRN在腎缺血／再灌注小鼠模型中具有腎臟保護(hù)作用。膿毒癥是急性腎損傷最主要的病因之一,而PGRN在膿毒癥誘導(dǎo)的急性腎損傷中的作用尚不明確。本文利用兩種常用的膿毒癥相關(guān)急性腎損傷研究模型：LPS腹腔注射誘導(dǎo)內(nèi)毒素血癥和盲腸結(jié)扎穿孔術(shù)誘導(dǎo)多細(xì)菌感染建立膿毒癥小鼠模型,初步探討PGRN在膿毒癥誘導(dǎo)的急性腎損傷中的功能。目的：明確PGRN在膿毒癥相關(guān)的急性腎損傷中的作用,并探討PGRN對(duì)膿毒癥相關(guān)腎損傷中腎組織炎癥應(yīng)答與腎小管上皮細(xì)胞死亡的影響。方法：分別使用小鼠腹腔注射細(xì)菌脂多糖(lipopolysaccharides, LPS)和小鼠盲腸結(jié)扎再穿孔(cecal ligation and puncture, CLP)的方法,構(gòu)建膿毒癥模型。在建模6小時(shí)和24小時(shí)后,使用實(shí)時(shí)定量PCR檢測(cè)腎組織中PGRN的mRNA水平,Western blot和免疫組化檢測(cè)PGRN的蛋白表達(dá),酶聯(lián)免疫吸附實(shí)驗(yàn)(enzyme-linked immunosorbent assay, ELISA)檢測(cè)血清與腎組織勻漿中PGRN的蛋白水平；在建模2小時(shí)前,野生型鼠外源性注射rPGRN和無(wú)菌PBS,其他野生型鼠和PGRN基因敲除鼠直接建模,為了檢測(cè)膿毒癥誘導(dǎo)的腎損傷的程度,血生化分析法檢測(cè)血清中肌酐和尿素氮含量,HE染色法檢測(cè)腎組織的損傷；免疫組化法檢測(cè)腎組織中中性粒細(xì)胞和巨噬細(xì)胞浸潤(rùn)；實(shí)時(shí)定量PCR法檢測(cè)腎組織炎性細(xì)胞因子的mRNA水平；TUNEL法檢測(cè)腎組織細(xì)胞的凋亡；Western blot檢測(cè)腎組織中凋亡蛋白的表達(dá)水平。結(jié)果：在兩種膿毒癥小鼠模型研究中,本研究發(fā)現(xiàn)膿毒癥小鼠腎組織中PGRN的蛋白水平和mRNA水平均高于對(duì)照組小鼠；與野生型鼠相比,PGRN基因敲除的膿毒癥小鼠的血清中肌酐和尿素氮水平更高；HE染色顯示腎組織損傷更加嚴(yán)重；中性粒細(xì)胞和巨噬細(xì)胞浸潤(rùn)更加嚴(yán)重；促炎細(xì)胞因子IL-6、TNF-α、IL-1p的mRNA水平升高；腎組織細(xì)胞凋亡更顯著,凋亡蛋白的表達(dá)水平升高；而與注射PBS組相比,外源性注射rPGRN組,小鼠腎功能障礙和腎組織損傷均明顯減輕；注射rPGRN能夠有效抑制腎組織中炎癥應(yīng)答及腎小管上皮細(xì)胞的凋亡性死亡。結(jié)論：PGRN在膿毒癥小鼠腎組織及外周血中表達(dá)水平升高；PGRN敲除導(dǎo)致膿毒癥相關(guān)急性腎損傷更加嚴(yán)重；而外源性注射重組PGRN后腎損傷明顯改善。本研究采用野生型與PGRN基因敲除小鼠或外源注射rPGRN等方法,利用LPS注射和CLP膿毒癥小鼠模型,明確了PGRN對(duì)膿毒癥相關(guān)急性腎損傷的保護(hù)性作用,并從炎癥應(yīng)答和腎臟實(shí)質(zhì)細(xì)胞死亡的角度分析了PGRN腎保護(hù)作用的機(jī)制,為臨床膿毒癥尤其是膿毒癥誘導(dǎo)的急性腎損傷的防治,提供了新的理論依據(jù)。
[Abstract]:In recent years , PGRN is one of the most common complications of sepsis , including fatty tissue , hematopoietic cells , chondrocytes and immune cells . ELISA was used to detect the level of PGRN protein in serum and renal tissue homogenate ;
In order to detect the degree of renal injury induced by sepsis , blood biochemical analysis was used to detect serum creatinine and urea nitrogen content and HE staining was used to detect renal tissue injury .
Immunohistochemical method was used to detect the infiltration of neutrophils and macrophages in renal tissue .
The mRNA level of inflammatory cytokines in renal tissue was measured by real - time quantitative PCR .
TUNEL method was used to detect the apoptosis of renal tissue cells .
The expression level of apoptotic protein in renal tissue was detected by Western blot . Results : In the study of two types of septic mice , the protein level and mRNA level of PGRN in renal tissue of septic mice were higher than those in control group .
Compared with wild - type mice , the level of creatinine and urea nitrogen was higher in the serum of septic mice with PGRN knockout mice .
HE staining showed that renal tissue injury was more serious .
The infiltration of neutrophils and macrophages was more serious ;
The mRNA levels of pro - inflammatory cytokines IL - 6 , TNF - 偽 and IL - 1p were increased .
Apoptosis of renal tissue cells was more significant , and the level of apoptosis protein was increased .
Compared with PBS group , the rPGRN group , renal dysfunction and renal tissue injury in mice were significantly reduced .
Conclusion : PGRN can effectively inhibit the inflammatory response in renal tissue and death of renal tubular epithelial cells . Conclusion : The expression of PGRN in renal tissue and peripheral blood of septic mice is increased .
PGRN knockout resulted in more severe acute renal injury associated with sepsis ;
In this study , the protective effects of PGRN on acute renal injury associated with sepsis were identified by using wild - type and PGRN knockout mice or exogenous injected rPGRN , and the mechanism of PGRN kidney protection was analyzed from the angle of inflammatory response and renal parenchyma cell death , which provided a new theoretical basis for the prevention and treatment of acute renal injury induced by sepsis , especially sepsis .

【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R459.7;R692.5

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 姚詠明,盛志勇;膿毒癥研究的若干新動(dòng)態(tài)[J];中國(guó)危重病急救醫(yī)學(xué);2000年06期

2 菲琳;;治療膿毒癥的新希望[J];國(guó)外醫(yī)學(xué)情報(bào);2002年10期

3 向陽(yáng);防止膿毒癥自高動(dòng)力相轉(zhuǎn)向低動(dòng)力相的新途徑[J];國(guó)外醫(yī)學(xué).外科學(xué)分冊(cè);2003年03期

4 任新生;重新認(rèn)識(shí)全身炎癥反應(yīng)綜合征、膿毒癥和多器官功能衰竭綜合征[J];中華急診醫(yī)學(xué)雜志;2004年02期

5 崔德健;探討膿毒癥診斷和治療新策略[J];中國(guó)呼吸與危重監(jiān)護(hù)雜志;2004年03期

6 林洪遠(yuǎn);膿毒癥診斷和治療進(jìn)展[J];中國(guó)實(shí)用外科雜志;2004年06期

7 湯耀卿;膿毒癥的定義和診斷[J];臨床外科雜志;2004年11期

8 王小平;膿毒癥研究現(xiàn)狀[J];實(shí)用臨床醫(yī)學(xué);2005年06期

9 周國(guó)勇;性別與膿毒癥嚴(yán)重程度的關(guān)系[J];中國(guó)危重病急救醫(yī)學(xué);2005年07期

10 黎永明;姜勇;;膿毒癥發(fā)生機(jī)制的新進(jìn)展[J];感染.炎癥.修復(fù);2005年01期

相關(guān)會(huì)議論文 前10條

1 張麗葳;奚希相;張威;張莉芬;陳昊;李俊;楊興易;;以血小板減少為主要表現(xiàn)的膿毒癥搶救一例[A];《中華急診醫(yī)學(xué)雜志》第八屆組稿會(huì)暨急診醫(yī)學(xué)首屆青年論壇論文匯編[C];2009年

2 張振輝;林s鉅，

本文編號(hào)：1854088