本文選題：盲腸結(jié)扎穿孔 + 急性腎損傷�。� 參考：《安徽醫(yī)科大學》2013年碩士論文

【摘要】：目的：急性腎損傷(acute kidney injury, AKI)是危重患者最常見的臨床問題之一,膿毒癥一直是AKI的首要原因,膿毒癥合并AKI患者的死亡率高達70%。因此,探討膿毒癥時腎臟的保護治療有著重要的臨床意義。黑皮質(zhì)素4型受體(melanocortin4receptor, MC4R)在攝食、能量代謝、性功能、心血管功能等方面起著重要調(diào)節(jié)作用。有文獻報道選擇性激動MC4R可以激活膽堿能抗炎通路,顯著抑制重癥急性胰腺炎大鼠全身炎癥進展,保護胰腺組織。當膿毒癥合并AKI時,能否通過選擇性激動MC4R,激活膽堿能抗炎通路對抗炎癥反應,實現(xiàn)對膿毒癥患者腎臟的保護作用尚未見報道。本課題采用盲腸結(jié)扎穿孔術(cecal ligation and puncture, CLP),建立膿毒癥大鼠模型,觀察膿毒癥對腎臟的影響,探討膿毒癥AKI動物模型的制備。在膿毒癥AKI大鼠模型的基礎上,通過選擇性激動MC4R進行干預,觀察對腎臟的影響,闡述選擇性激動MC4R對膿毒癥大鼠AKI的保護作用和機制,為臨床預防和治療膿毒癥合并AKI提供實驗基礎和理論依據(jù)。 方法：采用CLP法制備膿毒癥大鼠模型(1)實驗大鼠隨機分為假手術組(Sham組)和盲腸結(jié)扎穿孔組(CLP組),每組各18只。二組均在造模后設置6h、12h和24h三個時間點取材,檢測大鼠心率(heart rate, HR)和平均動脈壓(mean arterial blood perssuer, MABP);測定血清中腫瘤壞死因子-a(tumor necrosis factor-a, TNF-a)和白細胞介素-6(interleukin-6, IL-6)的表達水平及血清肌酐(creatine,CRE)和尿素氮(blood urea nitrogen, BUN)水平；觀察大鼠腎臟組織結(jié)構(gòu)的改變；檢測腎臟組織中高遷移率族蛋白B1(high mobility group box1protein, HMGB1) mRNA的表達水平和核因子-κB(nuclear factor-KB, NF-κB)的活化。(2)實驗大鼠隨機分假手術組(Sham組)、假手術+Ro27-3225治療組(Sham+Ro27組)、盲腸結(jié)扎穿孔組(CLP組)和盲腸結(jié)扎穿孔+Ro27-3225治療組(CLP+Ro27組)。每組各6只。四組均在造模后24h取材,檢測大鼠HR和MABP;測定血清CRE和BUN水平；觀察大鼠腎臟組織結(jié)構(gòu)的改變；檢測腎臟組織中HMGB1mRNA的表達水平和NF-κ3的活化。 結(jié)果：(1)與同時間點Sham組比較,CLP組6h和12h大鼠HR和MABP明顯升高(p0.01),24h MABP降低(p0.01)；CLP組6h和12h大鼠血清中TNF-a和IL-6表達水平明顯升高(p0.01),24h降至Sham組水平(p0.05)；CLP組6h、12h和24h大鼠血清中BUN水平升高(p0.05和p0.01),24h血清中CRE水平明顯升高(p0.01)；CLP組12h大鼠腎臟組織中HMGB1mRNA表達水平出現(xiàn)升高,24h明顯升高(p0.01)；CLP組6h大鼠腎臟組織NF-kB p65核陽性率升高,且隨病程延長,NF-kB p65核陽性率明顯升高(p0.01)。Sham組各組大鼠腎小體形態(tài)正常,腎小管上皮細胞未見腫脹；CLP組各組大鼠腎小體中血管球淤血、皺縮,腎小囊腔擴大,且隨病程延長,淤血、皺縮的血管球增多,并伴有腎小管上皮細胞腫脹。(2)Sham+Ro27組大鼠HR和MABP較Sham組明顯降低(p0.01),CLP組大鼠MABP低于Sham組(p0.01),CLP+Ro27組大鼠MABP較CLP組升高(p0.01)；CLP組大鼠血清BUN和CRE水平較Sham組明顯升高(p0.01),CLP+Ro27組大鼠血清BUN和CRE水平較CLP組明濕降低(p0.01)；CLP組大鼠腎臟組織中HMGB1mRNA的表達水平較Sham組明顯升高(p0.01)；CLP+Ro27組大鼠腎臟組織中HMGB1mRNA的表達水平較CLP組降低(p0.01)；CLP組大鼠腎臟組織NF-kB p65核陽性率較Sham組升高；CLP+Ro27組大鼠腎臟組織中NF-kB p65核陽性率較CLP組降低(p0.01)；Sham組和Sham+Ro27組大鼠腎小體形態(tài)正常,腎小管上皮細胞未見腫脹,CLP組大鼠腎小體中血管球淤血、皺縮,腎小囊腔擴大,伴有腎小管上皮細胞腫脹,CLP+Ro27組大鼠也可見腎小體中血管球淤血、皺縮和腎小囊腔擴大,但數(shù)量明顯少于CLP組,間質(zhì)充血、水腫及炎性細胞浸潤也有所改善。 結(jié)論：采用CLP方法能成功的復制出膿毒癥AKI大鼠模型,引起大鼠腎臟結(jié)構(gòu)和功能的改變,引起腎臟組織中HMGB1mRNA的表達水平升高和NF-kB p65核陽性率增加；選擇性激動MC4R對膿毒癥AKI大鼠腎臟的結(jié)構(gòu)和功能都有保護作用,并且抑制腎臟組織中HMGB1mRNA的表達水平升高和NF-κB p65核陽性率增加。
[Abstract]:Objective: acute kidney injury (AKI) is one of the most common clinical problems in critically ill patients. Sepsis is the primary cause of AKI. The mortality of patients with sepsis with AKI is as high as 70%., so the protective treatment of kidney in sepsis has an important clinical significance. The 4 type of melanin receptor (melanocortin4receptor,) MC4R) plays an important regulatory role in feeding, energy metabolism, sexual function and cardiovascular function. It is reported that selective excitation of MC4R can activate the cholinergic anti-inflammatory pathway, significantly inhibit the progression of severe acute pancreatitis in rats and protect the pancreatic tissue. When AKI is combined with sepsis, it can be activated by selectively activating MC4R and activating the bile. The protective effect of alkaline energy anti-inflammatory pathway against inflammatory reaction has not yet been reported. This subject uses cecal ligation and puncture (CLP) to establish a rat model of sepsis, to observe the effect of sepsis on the kidney, and to explore the preparation of AKI animal model in sepsis. In the AKI rat model of sepsis, the model of sepsis AKI rat model is made. On the basis of selective stimulation of MC4R, the effect of MC4R on the kidney was observed, and the protective effect and mechanism of selective excitant MC4R on septic rat AKI were described, and the experimental basis and theoretical basis were provided for the clinical prevention and treatment of sepsis with AKI.
Methods: the rat model of sepsis (1) was prepared by CLP method. The experimental rats were randomly divided into sham operation group (group Sham) and cecum ligation group (group CLP), each group was 18. The two groups were set up 6h, 12h and 24h at three time points, and the heart rate (heart rate, HR) and mean arterial pressure (mean arterial blood) were measured. The level of the expression of tumor necrosis factor -a (tumor necrosis factor-A, TNF-a) and interleukin -6 (interleukin-6, IL-6) in serum and the level of serum creatinine (creatine, CRE) and urea nitrogen (blood urea), and the changes of renal tissue structure in rats were observed and the high mobility group protein in renal tissue was detected. OUP box1protein, HMGB1) mRNA expression level and the activation of nuclear factor kappa B (nuclear factor-KB, NF- kappa B). (2) experimental rats were randomly divided into sham operation group (Sham group), sham operation +Ro27-3225 treatment group (Sham+Ro27 group), caecum ligation and perforation group and cecum ligation perforation treatment group. Each group was made up of four groups. After 24h, HR and MABP were measured and the levels of serum CRE and BUN were measured. The changes of renal tissue structure of rats were observed. The expression of HMGB1mRNA in renal tissue and the activation of NF- kappa 3 were detected.
Results: (1) compared with group Sham at the same time point, HR and MABP in CLP group 6h and 12h rats increased significantly (P0.01) and 24h MABP decreased (P0.01). The level of HMGB1mRNA expression in renal tissue of 12h rats in group CLP was increased and 24h increased significantly (P0.01), and the positive rate of NF-kB p65 in renal tissue of group 6h rats increased, and the positive rate of NF-kB p65 nucleus increased obviously with the course of disease, and the renal corpuscle of renal tubules was not normal in all groups of rats. Swelling, vascular bulging, shrinkage and enlargement of renal capsule cavity in the renal corpuscle of rats in group CLP, with the extension of the course, the increase of blood vessel and crumpled blood vessel, and the swelling of renal tubular epithelial cells. (2) the HR and MABP in the group Sham+Ro27 rats were significantly lower than those in the Sham group (P0.01), and the MABP in the group CLP was lower than that in the Sham group (P0.01), and the CLP+Ro27 group MABP was higher than that of the group. The level of serum BUN and CRE in the group CLP rats was significantly higher than that in the Sham group (P0.01). The level of serum BUN and CRE in the CLP+Ro27 group was lower than that in the CLP group (P0.01), and the expression level of the kidneys in the rats of the CLP group was significantly higher than that in the Sham group; the expression level of the kidney in the rats was lower than that in the group of rats. (P0.01); the positive rate of NF-kB p65 nucleus in kidney tissue of rats in group CLP was higher than that in group Sham, and the positive rate of NF-kB p65 in kidney tissue of group CLP+Ro27 was lower than that in group CLP (P0.01), and the renal corpuscle cells in Sham group and Sham+Ro27 group were normal, no swelling of renal tubular epithelial cells, vascular congestion, shrinkage and enlargement of renal cysts in renal corpuscles of rats. The epithelial cells of renal tubule were swollen, and the rats in group CLP+Ro27 also found vascular congestion, contraction and enlargement of the renal capsule cavity in the renal corpuscle, but the number was significantly less than that in the CLP group. The interstitial congestion, edema and inflammatory cell infiltration were also improved.
Conclusion: CLP can successfully replicate the AKI rat model of sepsis, cause changes in the structure and function of kidney in rats, increase the level of HMGB1mRNA expression in renal tissue and increase the positive rate of NF-kB p65 nucleus; selective excitant MC4R has protective effect on the structure and function of kidney in septic AKI rats, and inhibits the kidney. The expression level of HMGB1mRNA and NF- kappa B p65 were increased in tissues.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R459.7

【共引文獻】

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本文編號：2034886