本文選題：川崎病　切入點(diǎn)：抵抗素　出處：《重慶醫(yī)科大學(xué)》2013年博士論文

【摘要】：第一部分血清抵抗素水平與川崎病的相關(guān)性研究 目的：川崎病是一種急性發(fā)熱性疾病，主要發(fā)生在嬰幼兒及年齡小的兒童，常伴有冠狀動(dòng)脈損傷。由于15%-25%的患兒有發(fā)生冠狀動(dòng)脈瘤的危險(xiǎn)，所以近年來(lái)引起了兒科醫(yī)生的極大關(guān)注。抵抗素是新近發(fā)現(xiàn)的一種脂肪細(xì)胞來(lái)源的肽類，屬于一類富含半胱氨酸殘基的分泌性蛋白家族，有研究發(fā)現(xiàn)抵抗素可能參與了炎癥過(guò)程，且與冠脈動(dòng)脈鈣化，粥樣硬化等疾病的發(fā)生發(fā)展密切相關(guān)。因以，本研究探討血清抵抗素水平與川崎病患兒及其發(fā)生冠脈動(dòng)脈瘤的相關(guān)性。 方法:用酶聯(lián)免疫吸附試驗(yàn)檢測(cè)了165例兒童血清抵抗素水平，并將其分為4組：健康對(duì)照組（n＝85），無(wú)冠脈動(dòng)脈損傷的川崎病患兒組（N=41），，伴冠脈擴(kuò)張的川崎病患兒組（N=31），并發(fā)冠狀動(dòng)脈瘤的川崎病患兒組（N=8）。同時(shí)統(tǒng)計(jì)其血液學(xué)指標(biāo),包括血白細(xì)計(jì)數(shù)（WBC），紅細(xì)胞計(jì)數(shù)（RBC），血紅蛋白水平（HB），紅細(xì)胞壓積（Hct），血小板計(jì)數(shù)，C-反應(yīng)蛋白（CRP）和紅細(xì)胞沉降率（ESR）。 結(jié)果:與正常對(duì)照組及川崎病患兒其他組相比，伴有冠肪動(dòng)脈瘤的川崎病患兒血清抵抗素水平明顯的升高，此外血紅蛋白水平明顯降低；且在患兒組，血清抵抗素水平與其CRP水平呈顯著正相關(guān)關(guān)系，而與患兒紅細(xì)胞水平呈負(fù)相關(guān)。 結(jié)論:抵抗素和血紅蛋白可能與川崎病患兒冠狀動(dòng)脈瘤的發(fā)生有關(guān)，血清抵抗素的表達(dá)上調(diào)可能與川崎病急性期的炎癥因子增加有關(guān)。第二部分抵抗素基因啟動(dòng)子-420C/G多態(tài)性與兒童川 崎病的相關(guān)性研究 目的：川崎�。↘D）是一種小兒全身性血管炎，通常伴發(fā)冠狀動(dòng)脈病變（CAL）。抵抗素為近年來(lái)發(fā)現(xiàn)的一種脂肪細(xì)胞來(lái)源的肽類，可能與炎癥反應(yīng)及冠脈動(dòng)脈疾病的發(fā)生密切相關(guān)。-420C G多態(tài)性位點(diǎn)位于抵抗素基因（RETN）啟動(dòng)子區(qū)域，新近研究顯示該區(qū)位點(diǎn)多態(tài)性在炎癥發(fā)生及心血管疾病中起到了重要作用。然而，RETN啟動(dòng)子多態(tài)性與川崎病之間的關(guān)聯(lián)尚不清楚。故本研究擬探討抵抗素基因多態(tài)性（－420C/G）與兒童川崎病易感性及其臨床特征的相關(guān)性。 方法：共計(jì)91名川崎病患兒，115名正常對(duì)照組兒童納入本研究。依照診斷標(biāo)準(zhǔn)將川崎病患兒分為了完全性（cKD）和不完全性川崎�。╥KD）。采用基質(zhì)輔助激光解吸電離飛行時(shí)間質(zhì)譜技術(shù)(MALDI-TOF)進(jìn)行抵抗素啟動(dòng)子基因多態(tài)性分型。采用夾心酶聯(lián)免疫法（ELISA）對(duì)所有血清樣本抵抗素蛋白濃度進(jìn)行測(cè)定。 結(jié)果：KD組和健康對(duì)照組抵抗素基因（-420CG）的基因型和其等位基因頻率進(jìn)行了比較，然而差異無(wú)統(tǒng)計(jì)學(xué)意義（P〈0.05）。此外，該基因位點(diǎn)多態(tài)性與川崎病的CALS的發(fā)生也無(wú)明顯差異。然而，G等位基因的頻率在iKD組明顯高于cKD組（P＜0.05），GG基因型也在iKD組顯著增加（P＜0.05）。KD組血清抵抗素水平明顯高于對(duì)照組（P＜0.05）。與基因型CC和CG和相比，GG基因型KD患兒血清抵抗素水平較高，但差異無(wú)統(tǒng)計(jì)學(xué)意義。 結(jié)論：本研究結(jié)果表明，抵抗素可能在KD的發(fā)病機(jī)制中發(fā)揮作用，位于RETN基因啟動(dòng)子的-420CG基因多態(tài)性與兒童川崎病無(wú)明顯相關(guān),然而該位點(diǎn)等位基因G及GG基因型有助于不完全性川崎病的診斷。第三部分MAPKs信號(hào)通路在介導(dǎo)抵抗素致人冠狀動(dòng)脈 內(nèi)皮細(xì)胞分泌MMP-9、TIMP-1中的作用研究 目的：抵抗素為脂肪細(xì)胞特異性分泌因子，可能與炎癥和心血管疾病的發(fā)生相關(guān)。本課題組前期研究表明血清抵抗素水平在川崎病患兒明顯升高，并與冠狀動(dòng)脈瘤的發(fā)生密切相關(guān)；但其致冠脈損傷的機(jī)制尚不清楚�；|(zhì)金屬蛋白酶-9（MMP-9）在KD患兒的炎癥反應(yīng)及血管損傷中起到了關(guān)鍵作用。因此，本課題提出假設(shè)，抵抗素可能對(duì)內(nèi)皮細(xì)胞MMP-9及金屬蛋白酶組織抑制劑（TIMP-1）的有調(diào)控作用，并進(jìn)行機(jī)制的探討。 方法:用人重組抵抗素處理人冠狀動(dòng)脈內(nèi)皮細(xì)胞（HCAECs）。用RT-PCR方法檢測(cè)細(xì)胞MMP-9和TIMP-1的表達(dá)，用ELISA法測(cè)定MMP-9和TIMP-1蛋白表達(dá)。用Western印跡分析ERK1/2,P38MAPK,JNK的總蛋白及磷酸化蛋白表達(dá)。 結(jié)果：抵抗素可刺激冠脈內(nèi)皮細(xì)胞MMP-9的分泌和減少其TIMP-1的產(chǎn)生,并有時(shí)間和濃度依賴性。ERK1/2抑制劑（U0126），p38MAPK抑制劑(SB202190），和JNK抑制劑（SP600125）可明顯減少抵抗素致HCAECsMMP-9的分泌。 結(jié)論：綜上所述，ERK1/2，p38和JNK信號(hào)通路可能參與了抵抗素致HCAECs MMP-9的分泌。MAPKs信號(hào)通路可能與抵抗素相關(guān)的心血管疾病有關(guān)。
[Abstract]:The first part of study on correlation between serum resistin levels and Kawasaki disease
Objective: Kawasaki disease is an acute febrile illness occurred mainly in infants and children younger, often accompanied by coronary artery injury. Due to 15%-25% with risk of coronary artery aneurysms, so in recent years has aroused great concern to pediatricians. Resistin is an adipocyte derived peptide discovered recently, belongs to a class of cysteine rich residues of secretory protein family, studies have found that resistin may be involved in the inflammatory process, and coronary artery calcification, closely related to the occurrence and development of atherosclerosis and other diseases. Due to this research, the correlation between serum resistin level and coronary artery disease in children and Kawasaki tumors.
Methods: 165 cases of children with serum resistin levels were detected by enzyme-linked immunosorbent assay, and divided into 4 groups: normal control group (n = 85), without coronary artery injury in children with Kawasaki group (N=41), with the expansion of coronary artery disease in children Kawasaki group (N=31), coronary artery aneurysm Kawasaki disease group (N=8). At the same time, the statistics of hematological indexes, including blood leukocyte count (WBC), red blood cell count (RBC), hemoglobin (HB), hematocrit (Hct), platelet count, C- reaction protein (CRP) and erythrocyte sedimentation rate (ESR).
Results: compared with normal control group and Kawasaki group with other disease patients, aneurysms Kawasaki crown fat disease in children serum resistin levels significantly increased, in addition to hemoglobin levels decreased significantly; and in group, was significant positive correlation between serum resistin levels and the CRP level, but negatively correlated with the level of red blood cells in children.
Conclusion: Resistin and hemoglobin may be related to the occurrence of coronary artery aneurysm disease Kawasaki, increased serum resistin expression may be related to the increase of inflammatory factors in acute stage of Kawasaki. The second part of the resistin gene promoter -420C/G polymorphism and Sichuan children
Study on the correlation of kazaki disease
Objective: Kawasaki disease (KD) is a systemic vasculitis, usually associated with coronary artery disease (CAL) peptide. A fat cell source for resistin found in recent years, may be related to inflammation and coronary artery disease is closely related to the occurrence of.-420C G polymorphisms in resistin gene (RETN) promoter recent research shows that this region polymorphism plays an important role in cardiovascular disease and inflammation. However, the association between RETN promoter polymorphism and sub Kawasaki disease remains unclear. Therefore, this study intends to explore the resistin gene polymorphism (420C/G) and its correlation with clinical characteristics of children with Kawasaki disease susceptibility.
Methods: a total of 91 Kawasaki disease in children, 115 normal controls were included in the study group of children. Children to complete Kawasaki disease according to the diagnostic criteria (cKD) and incomplete Kawasaki disease (iKD). By using matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF) of resistin gene promoter polymorphism points type. Using a sandwich enzyme immunoassay (ELISA) for all serum samples of resistin protein concentrations were measured.
Results: KD group and healthy control group (-420CG) of resistin gene genotype and allele frequencies were compared, but the difference was not statistically significant (P < 0.05). In addition, there is no obvious difference between CALS gene polymorphism and the Kawasaki disease occurred. However, the frequency of G allele in the iKD group was significantly higher than that of group cKD (P < 0.05), the GG genotype was significantly increased in iKD group (P < 0.05) resistin level of serum.KD was significantly higher than the control group (P < 0.05). And the genotypes of CC and CG and compared the serum resistin level in children with KD GG genotype is higher, but no significant the difference in meaning.
Conclusion: the results of this study show that resistin may play a role in the pathogenesis of KD, located in RETN gene promoter -420CG gene polymorphism and childhood disease was not related to Kawasaki, however, the allele G and genotype GG were helpful to the diagnosis of incomplete Kawasaki disease. The third part of the MAPKs signaling pathway in resistin mediated by coronary artery
The role of endothelial cells in the secretion of MMP-9 and TIMP-1
Objective: to resistin adipocyte specific secretory factors that may be associated with inflammation and cardiovascular disease. Previous study showed that serum resistin levels were significantly increased in patients with coronary artery disease and Kawasaki, tumor is closely related to the occurrence of the damage caused by coronary artery; but the mechanism is not clear. -9 (matrix metalloproteinase MMP-9) in inflammation and vascular injury in children with KD to play a key role. Therefore, this paper put forward the hypothesis of resistin on endothelial cell MMP-9 and tissue inhibitor of metalloproteinases (TIMP-1) are used as control, and explore the mechanism.
Methods: human coronary artery endothelial cells (HCAECs) were treated with recombinant human resistin. The expression of MMP-9 and TIMP-1 was detected by RT-PCR. The expression of MMP-9 and TIMP-1 protein was detected by ELISA. The expression of total protein and phosphorylated protein in ERK1/2, P38MAPK and JNK were analyzed by Western blot.
Results: Resistin stimulated the secretion of MMP-9 and decreased the production of TIMP-1 in coronary endothelial cells, and there were time and concentration dependent.ERK1/2 inhibitors (U0126), p38MAPK inhibitors (SB202190), and JNK inhibitors (SP600125), which significantly reduced the secretion of resistin induced HCAECsMMP-9.
Conclusion: In conclusion, ERK1/2, p38 and JNK signaling pathways may be involved in resistin induced HCAECs MMP-9 secretion, and.MAPKs signaling pathway may be related to resistin related cardiovascular diseases.

【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2013
【分類號(hào)】：R725.4

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 ;Relationship between resistin level in serum and acute coronary syndrome or stable angina pectoris[J];Journal of Zhejiang University(Science B:An International Biomedicine & Biotechnology Journal);2007年12期

本文編號(hào)：1655012