本文關(guān)鍵詞：上皮細(xì)胞—樹突狀細(xì)胞轉(zhuǎn)分化在失血性休克誘發(fā)SIRS中的作用及維生素C的保護(hù)機(jī)制研究　出處：《上海交通大學(xué)》2015年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 失血性休克 全身炎癥反應(yīng)綜合征 維生素C 樹突狀細(xì)胞特異性細(xì)胞間粘附分子非整合素蛋白-3 糖原合成酶激酶-3β

【摘要】：目的:探討上皮細(xì)胞-樹突狀細(xì)胞轉(zhuǎn)分化(EDT)在失血性休克誘發(fā)全身炎癥反應(yīng)綜合征(SIRS)中的作用及可能機(jī)制;并進(jìn)一步研究維生素C是否通過抑制上皮細(xì)胞-樹突狀細(xì)胞轉(zhuǎn)分化而抑制失血性休克導(dǎo)致的大鼠多器官損傷。方法:以缺氧培養(yǎng)干預(yù)體外培養(yǎng)的大鼠腸粘膜上皮IEC-6細(xì)胞,部分給予維生素C干預(yù)(20-100uM)。在缺氧培養(yǎng)2、6、24、48小時(shí)后提取蛋白質(zhì)、mRNA及培養(yǎng)上清液。觀察樹突狀細(xì)胞特異性細(xì)胞間粘附分子非整合素蛋白3(DC-SIGN)、E-鈣粘蛋白(E-cadherin)、糖原合成酶激酶-3(GS K-3β)表達(dá)情況,及上清液白介素-1β(IL-1β)、白介素-6(IL-6)含量變化;從表型及分泌功能上探討缺氧對(duì)小腸上皮細(xì)胞-樹突狀細(xì)胞轉(zhuǎn)分化的影響,及VitC對(duì)這一過程所起的作用(n=3);以siRNA干擾IEC-6的E-cadherin表達(dá),比較正常IEC-6和經(jīng)過siR NA干擾的IEC-6在缺氧培養(yǎng)及Vit C干預(yù)后DC-SIGN表達(dá)情況的不同;以TDZD-8抑制GSK-3β活性,比較正常IEC-6和經(jīng)過TDZD-8干預(yù)的IEC-6在缺氧培養(yǎng)及VitC干預(yù)后,DC-SIGN、E-cadherin表達(dá)情況的不同。共同探討缺氧誘導(dǎo)上皮細(xì)胞-樹突狀細(xì)胞轉(zhuǎn)分化的可能機(jī)制(n=3)。通過股動(dòng)脈放血建立大鼠失血性休克模型,部分大鼠在復(fù)蘇時(shí)給予維生素c干預(yù)(100mg/kg)。于復(fù)蘇后2、6、24小時(shí)處死大鼠取標(biāo)本,檢測(cè)小腸、腎臟組織內(nèi)dc-sign表達(dá)情況及定位,組織病理?yè)p傷,血漿和組織內(nèi)促炎細(xì)胞因子含量(腫瘤壞死因子-a、白介素-6),及血清生化指標(biāo)(尿素氮、肌酐、乳酸脫氫酶及乳酸)(n=6)。探討失血性休克狀態(tài)下的上皮細(xì)胞-樹突狀細(xì)胞轉(zhuǎn)分化現(xiàn)象與組織炎癥反應(yīng)、臟器病理及功能損害的關(guān)系,及vitc干預(yù)所起的作用;結(jié)果:缺氧培養(yǎng)2小時(shí)后iec-6細(xì)胞表達(dá)dc-sign顯著升高,同時(shí)e-cadherin表達(dá)下降,gsk-3β活性下降(p0.05)。維生素c以劑量依賴的方式抑制缺氧誘導(dǎo)的iec-6細(xì)胞dc-sign表達(dá)升高,并緩解缺氧導(dǎo)致的e-cadherin表達(dá)下降及gsk-3β活性下降(p0.05)。預(yù)先使用sirna干擾e-cadherin表達(dá)后,維生素c抑制缺氧誘導(dǎo)iec-6細(xì)胞dc-sign表達(dá)升高的作用被扭轉(zhuǎn)(p0.05)。預(yù)先使用特異性gsk-3β活性抑制劑tdzd-8抑制gsk-3β活性后,維生素c抑制缺氧誘導(dǎo)iec-6細(xì)胞e-cadherin表達(dá)下降的作用被扭轉(zhuǎn)(p0.05)。失血性休克誘導(dǎo)小腸上皮細(xì)胞、腎小管上皮細(xì)胞表達(dá)dc-sign,維生素c(100mg/kg)顯著抑制dc-sign的表達(dá)(p0.05)。維生素c干預(yù)顯著抑制小腸、腎臟病理?yè)p傷,降低組織及血漿促炎細(xì)胞因子水平,改善血清生化指標(biāo)(p0.05)結(jié)論:缺氧培養(yǎng)及失血性休克可以誘導(dǎo)上皮細(xì)胞-樹突狀細(xì)胞轉(zhuǎn)分化;維生素c對(duì)這一現(xiàn)象有抑制作用,并可通過抑制上皮細(xì)胞-樹突狀細(xì)胞轉(zhuǎn)分化有效減輕失血性休克早期多器官炎癥反應(yīng)。
[Abstract]:Objective: To investigate the epithelial cells transdifferentiation of dendritic cells (EDT) in hemorrhagic shock induced systemic inflammatory response syndrome (SIRS) in the role and possible mechanism; and further study whether vitamin C through inhibition of epithelial cells and dendritic cell differentiation and inhibition of hemorrhagic shock induced multiple organ injury of rats. Methods: the rat intestinal mucosal epithelial IEC-6 cells were cultured in vitro, and the vitamin C intervention (20-100uM) was partially given. Protein, mRNA and culture supernatant were extracted after 2, 6, 24 and 48 hours in hypoxia. Observation of dendritic cell specific intercellular adhesion molecule 3 (DC-SIGN) non integrin protein, E- cadherin (E-cadherin) and glycogen synthase kinase -3 (GS K-3) expression, and supernatants of interleukin -1 beta (IL-1 beta) and interleukin -6 (IL-6) content change; from the table type and secretory function on effect hypoxia on the transdifferentiation of intestinal epithelial cells, dendritic cells, and the VitC of the process effect (n=3); siRNA interference IEC-6 expression of E-cadherin, IEC-6 and siR compared to normal after NA interference of IEC-6 intervention DC-SIGN in hypoxic culture and Vit expression of C is different; inhibition of GSK-3 beta activity in TDZD-8 compared with normal IEC-6 and TDZD-8, after the intervention of IEC-6 in hypoxia and the intervention of VitC, DC-SIGN and E-cadherin expression in different situations. To discuss the possible mechanism of hypoxia induced transdifferentiation of epithelial cells - dendritic cells (n=3). The rat model of hemorrhagic shock was established through the blood release of the femoral artery, and some rats were given vitamin C intervention (100mg/kg) during the resuscitation. In 2, after the recovery of 6, 24 hours, the rats were sacrificed, specimens, detection of the small intestine and kidney tissue DC-SIGN expression and localization, tissue damage, inflammatory cytokines in plasma and tissues (promoting tumor necrosis factor interleukin -a, -6), and serum biochemical indexes (blood urea nitrogen, creatinine, lactate dehydrogenase and lactic acid) (n=6). To investigate the relationship between hemorrhagic shock under the condition of epithelial cells and dendritic cell transdifferentiation and tissue inflammation, organ function and pathological damage, and the effect of VitC intervention; results: 2 hours after hypoxia in IEC-6 cells the expression of DC-SIGN was significantly increased, while expression of E-cadherin decreased, decreased GSK-3 activity (P0.05). Vitamin C inhibited the expression of DC-SIGN in hypoxia induced IEC-6 cells in a dose-dependent manner, alleviated the decrease of E-cadherin expression and decreased the activity of GSK-3 beta (P0.05). When siRNA interfered with E-cadherin expression in advance, the effect of vitamin C on the inhibition of hypoxia induced DC-SIGN expression in IEC-6 cells was reversed (P0.05). After the specific GSK-3 beta activity inhibitor tdzd-8 was used to inhibit GSK-3 beta activity, vitamin C inhibited the decrease of E-cadherin expression in IEC-6 cells induced by hypoxia, which was reversed (P0.05). The expression of DC-SIGN in small intestinal epithelial cells and renal tubular epithelial cells was induced by hemorrhagic shock, and vitamin C (100mg/kg) significantly inhibited the expression of DC-SIGN (P0.05). Vitamin C supplementation significantly inhibited intestine and kidney pathological damage, reduce tissue and plasma levels of proinflammatory cytokines, improve the serum biochemical indexes (P0.05) conclusion: hypoxia and hemorrhagic shock can induce epithelial cell dendritic cell transdifferentiation; vitamin C on this phenomenon are inhibited, and through inhibition of epithelial cells dendritic cell transdifferentiation can effectively reduce the blood loss of multiple organ inflammatory reaction at early stage of shock.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R459.7

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