本文選題：高分子量激肽原 + 緩激肽　； 參考：《蘇州大學(xué)》2014年碩士論文

【摘要】：【研究背景和意義】 血漿高分子量激肽原（High-molecular-weight kininogen，HK）是血漿激肽釋放酶-激肽系統(tǒng)（kallikrein-kinin system，KKS）的主要成員之一。臨床研究表明，IBD（inflammatory bowel disease）患者體內(nèi)血漿KKS被活化，HK被裂解，說(shuō)明HK參與IBD的發(fā)病過(guò)程。但是，迄今對(duì)于HK在IBD發(fā)病中的確切作用和機(jī)制并不清楚。 【研究目的】 研究血漿高分子量激肽原（HK）在葡聚糖硫酸鈉（Dextran sulfate sodium，DSS）誘導(dǎo)的急性結(jié)腸炎中的作用，并初步探究其作用機(jī)制。 【研究方法】 用含2.5%DSS的飲用水飼喂小鼠，構(gòu)建急性結(jié)腸炎模型。通過(guò)檢測(cè)體重百分比，疾病活動(dòng)指數(shù)，結(jié)腸長(zhǎng)度與腸系膜淋巴結(jié)的濕重比較野生型小鼠和Kng1基因敲除小鼠發(fā)病程度。利用ELISA方法檢測(cè)血漿緩激肽（BK），結(jié)腸組織中炎癥因子與髓過(guò)氧化物酶（MPO）的含量。應(yīng)用實(shí)時(shí)定量PCR檢測(cè)結(jié)腸組織炎癥因子的mRNA水平。分離小鼠結(jié)腸粘膜固有層細(xì)胞，通過(guò)特異性的抗體標(biāo)記后用流式細(xì)胞儀檢測(cè)細(xì)胞懸液中中性粒細(xì)胞所占的比例。制作末端結(jié)腸的石蠟切片，通過(guò)HE染色觀察結(jié)腸組織病理變化。 【研究結(jié)果】 （1）Kng1基因敲除小鼠肝臟中沒(méi)有Kng1mRNA的表達(dá)，并且血漿中檢測(cè)不到HK蛋白，說(shuō)明Kng1基因敲除小鼠模型建立成功。（2）DSS處理的WT小鼠血漿BK水平升高。（3）從day0起，給予小鼠DSS處理。與對(duì)照組小鼠相比，DSS處理組小鼠從day4開始體重逐漸下降。與WT小鼠相比，DSS處理的Kng1-/-小鼠體重下降程度（day5: p=0.0343; day6: p=0.024; day7: p=0.0248; day8: p=0.019）和疾病活動(dòng)指數(shù)（day7: p=0.0331; day8：p=0.0039）明顯降低，其結(jié)腸縮短程度明顯減輕（p=0.021），而且其腸系膜淋巴結(jié)的濕重明顯低于WT小鼠（p=0.0002）。（4）與野生型發(fā)病小鼠相比，DSS處理的Kng1-/-小鼠結(jié)腸組織中炎癥因子的蛋白水平明顯降低（TNF-α：p=0.032；IL-6：p=0.0083；IL-1β：p=0.0143；INF-γ：p=0.0012)，其結(jié)腸組織中炎癥因子的mRNA水平明顯降低（TNF-α：p=0.083；IL-6：p=0.0031；IL-1β：p=0.0036；INF-γ：p=0.024）（5）與野生型發(fā)病小鼠相比，DSS處理的Kng1-/-小鼠結(jié)腸組織研磨液中髓過(guò)氧化酶的含量顯著降低（p=0.001），，其結(jié)腸粘膜固有層中性粒細(xì)胞的數(shù)目顯著降低（p=0.002）。（6）野生型發(fā)病小鼠結(jié)腸粘膜上皮潰瘍，大量炎性細(xì)胞浸潤(rùn)，隱窩膿腫，杯狀細(xì)胞消失。與之相比，DSS處理的Kng1基因敲除小鼠結(jié)腸組織損傷與病變程度明顯緩解，結(jié)腸粘膜上皮僅可見部分破損，少量炎癥細(xì)胞浸潤(rùn)，部分杯狀細(xì)胞破損，組織切片的病理評(píng)分低于WT小鼠（p=0.0026）。 【研究結(jié)論】 血漿高分子量激肽原在炎癥性腸病的發(fā)病中起重要的促進(jìn)作用。
[Abstract]:[background and significance of the study] High-molecular-weight kininogeny (HKK) is one of the major members of the plasma kallikrein-kinin system (KKS). Clinical studies showed that plasma KKS was activated by HK in patients with inflammatory bowel disease, which indicated that HK was involved in the pathogenesis of IBD. However, the exact role and mechanism of HK in the pathogenesis of IBD is unclear. [purpose of research] To study the role of plasma high molecular weight kallikrein (HKHK) in acute colitis induced by dextran sodium sulfate (Dextran sulfate sodium), and to explore its mechanism. [research methods] Acute colitis model was established by feeding mice with drinking water containing 2.5%DSS. The percentage of body weight disease activity index colon length and wet weight of mesenteric lymph nodes were compared between wild-type mice and Kng1 knockout mice. The content of inflammatory factors and myeloperoxidase (MPO) in colon tissue and plasma bradykinin (BK) were determined by ELISA method. Real-time quantitative PCR was used to detect the mRNA level of inflammatory factors in colon tissue. Murine colonic mucosal lamina propria cells were isolated and the proportion of neutrophils in suspension was detected by flow cytometry after specific antibody labeling. The paraffin sections of the terminal colon were made and the pathological changes of the colon were observed by HE staining. [results of research] There was no Kng1mRNA expression in the liver of Kng1 knockout mice, and no HK protein was detected in plasma, indicating that the plasma BK level of WT mice treated with Kng1 gene knockout mice was higher than that of WT mice treated with DSS. Compared with the control group, the mice in DSS group gradually lost weight from day4. Compared with WT mice, the weight loss degree of Kng1-r- mice treated with DSS was significantly lower than that of WT mice (day5: p0. 034; day6: p0. 024; day7: p0. 024; day8: p0. 019), and the disease activity index was 0. 033 1; day8: p0. 0039). Compared with wild-type mice treated with DSS, the level of inflammatory factor protein in colonic tissue of DSS treated with DSS significantly decreased the protein level of inflammatory factor in colonic tissue of TNF- 偽 -03032 IL-6: p0.0083IL-1 尾: p0.0143INF- 緯 -P0.0012.The colonic shortening degree of TNF- 偽 decreased significantly the wet weight of mesenteric lymph nodes in WT mice compared with that of wild-type mice treated with DSS, and the protein level of inflammatory factors in colonic tissue of mice treated with DSS was significantly lower than that of wild-type mice. MRNA level of inflammatory cytokines in intestinal tissue decreased significantly (TNF- 偽: 0. 083) IL-6: P0. 0031, IL-1 尾: p0. 0036 + INF- 緯: p0. 024. 5) compared with wild-type mice, the content of myeloperoxidase in the colonic tissue grinding fluid treated with DSSs significantly decreased the number of neutrophils in the mucosal lamina propria of the colonic mucosa, and the number of neutrophilic granulocytes in the mucosal lamina propria of the colonic mucosa. The colonic mucosal epithelium ulcer of wild-type mice was decreased significantly. A large number of inflammatory cell infiltration, crypt abscess, goblet cells disappear. Compared with the Kng1 gene knockout mice treated with DSS, the degree of colonic tissue injury and lesion was significantly alleviated. The colonic mucosal epithelium was only partially damaged, a small number of inflammatory cells infiltrated, and some goblet cells were damaged. The pathological score of histopathological sections was lower than that of WT mice. [conclusions of the study] Plasma high molecular weight kallikrein plays an important role in the pathogenesis of inflammatory bowel disease.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R574.62

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 中國(guó)炎癥性腸病協(xié)作組;王玉芳;歐陽(yáng)欽;;3100例潰瘍性結(jié)腸炎住院病例回顧分析[J];中華消化雜志;2006年06期

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