發(fā)布時間：2018-05-14 00:15

  本文選題：CD26 + 血管內(nèi)皮細胞��； 參考：《遵義醫(yī)學院》2015年碩士論文

【摘要】：目的:觀察高糖培養(yǎng)下人臍靜脈內(nèi)皮細胞(HUVEC)的CD26表達變化、甘露聚糖結(jié)合凝集素(MBL)補體途徑的激活及炎癥因子(IL-6和NF-k B)的表達情況,探討CD26在糖尿病腎病(DN)慢性炎癥發(fā)病機制中的作用,為CD26抑制劑治療DN尋求基礎(chǔ)研究依據(jù)。方法:體外培養(yǎng)人臍靜脈內(nèi)皮細胞,分別設(shè)置正常對照組(含葡萄糖5.5mmol/L),甘露醇組(含葡萄糖5.5mmol/L+甘露醇24.5mmol/L),高糖組(含葡萄糖30mmol/L),采用免疫組織化學法檢測細胞表面CD26、MBL、C3和膜攻擊復合物(MAC)的表達,采用Western Blot檢測細胞的CD26、MBL、C3及MAC的蛋白表達,采用實時熒光定量PCR(real-time PCR)檢測細胞中CD26、IL-6和NF-k B m RNA的表達情況。結(jié)果:⑴免疫組化提示:在HUVEC中CD26、MBL、C3、膜攻擊復合物(MAC)主要表達于胞膜和胞漿。高糖組CD26、MBL、C3及MAC的蛋白表達均明顯高于正常組與甘露醇組(P0.05);在正常組與甘露醇組之間,各指標蛋白表達量均無統(tǒng)計學意義(P0.05)。CD26與MBL、C3、MAC蛋白表達量均呈明顯正相關(guān)(r=0.874,r=0.952,r=0.947,P0.01)。⑵Western Blot提示:高糖組CD26、MBL、C3及MAC的蛋白表達亦明顯高于正常組及甘露醇組(P0.05);在正常組與甘露醇組之間,各指標蛋白表達量均無統(tǒng)計學意義(P0.05)。CD26與MBL、C3、MAC蛋白表達量呈明顯正相關(guān)(r=0.825,r=0.869,r=0.828,P0.01)。⑶RT-PCR提示:高糖組CD26、IL-6、NF-k B的m RNA表達均高于正常組及甘露醇組(P0.05);在正常組與甘露醇組之間,各指標m RNA表達量均無統(tǒng)計學意義(P0.05)。CD26與IL-6、NF-k B的m RNA表達量均呈正相關(guān)(r=0.858,r=0.794,P0.05)。結(jié)論:體外培養(yǎng)的人臍靜脈內(nèi)皮細胞可少量表達CD26。高糖條件下,HUVEC的CD26表達增高,后者可能通過激活MBL介導的甘露糖結(jié)合凝集素補體途徑,引起下游細胞因子IL-6、NF-k B表達增加,加重高糖作用下HUVEC的炎癥反應(yīng)。
[Abstract]:Aim: to observe the changes of CD26 expression in human umbilical vein endothelial cells (HUVECs), the activation of mannan-binding lectin (MBL) complement pathway and the expression of inflammatory cytokines (IL-6 and NF-k B) in cultured human umbilical vein endothelial cells (HUVECs). To explore the role of CD26 in the pathogenesis of chronic inflammation in diabetic nephropathy (DN), and to seek the basis for the treatment of DN with CD26 inhibitor. Methods: human umbilical vein endothelial cells were cultured in vitro. The expression of CD26MMBL3 and membrane attack complex (MAC-1) was detected by immunohistochemistry in normal control group (including glucose 5.5 mmol 路L ~ (-1), mannitol group (containing glucose 5.5mmol/L _ (2.5) mmol 路L ~ (-1) and high glucose group (containing glucose 30 mmol / L ~ (-1). The expression of CD26ML-C _ 3 and MAC was detected by Western Blot, and the expression of IL-6 and NF-k B m RNA was detected by real-time fluorescence quantitative PCR(real-time. Results Immunohistochemistry showed that CD26ML-C3, a membrane attack complex, was mainly expressed in the membrane and cytoplasm of HUVEC. The protein expressions of CD26ML-C _ 3 and MAC in high glucose group were significantly higher than those in normal group and mannitol group (P 0.05), and between normal group and mannitol group. There was no significant difference in the protein expression of P0.05G, CD26 and MBLL-C3C3MAC protein expression. The results showed that there was a significant positive correlation between the protein expression of P0.05, CD26 and MBC, and the expression of CD26ML-C3 and MAC in high glucose group was significantly higher than that in normal group and mannitol group, and between normal group and mannitol group, the protein expression of CD26ML-C _ 3 and MAC in mannitol group was significantly higher than that in normal group, while in mannitol group, the expression of CD26ML-C _ 3 and MAC in high glucose group was significantly higher than that in normal group and mannitol group. There was no statistical significance between the expression of P0.05U. CD26 and the MAC protein expression of MBLML-C3C3. The results of RT-PCR indicated that the expression of m RNA in high glucose group was higher than that in normal group and mannitol group, and between normal group and mannitol group, the expression of m RNA of NF-kB in high glucose group was higher than that in normal group and mannitol group, and in mannitol group, the expression of m RNA in high glucose group was significantly higher than that in normal group and mannitol group. There was no statistical significance between the expression of m RNA and the expression of m RNA in IL-6 nF-kB. There was a positive correlation between the expression of m RNA and the expression of m RNA in IL-6 nF-kB. Conclusion: in vitro cultured human umbilical vein endothelial cells can express a small amount of CD 26. The expression of CD26 in HUVEC was increased under high glucose condition. The latter may increase the expression of IL-6NF-K B by activating the MBL mediated mannose binding lectin complement pathway and aggravate the inflammatory response of HUVEC under high glucose.
【學位授予單位】：遵義醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R587.2;R692.9

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本文編號：1885457