發(fā)布時(shí)間：2018-06-13 12:50

  本文選題：組胺H2受體 + 組胺H4受體��； 參考：《第二軍醫(yī)大學(xué)》2014年碩士論文

【摘要】：研究目的 1、比較過(guò)敏性紫癜患兒急性期、恢復(fù)期及健康體檢兒童外周血單個(gè)核細(xì)胞表面組胺H2、H4受體的表達(dá)差異,探討組胺H2、H4受體與過(guò)敏性紫癜的關(guān)系。 2、比較過(guò)敏性紫癜非腎炎組、過(guò)敏性紫癜腎炎組組胺H2、H4受體表達(dá)的差異,了解組胺受體與過(guò)敏性紫癜腎炎的相關(guān)性。 3、比較予組胺H2受體拮抗劑西咪替丁治療前后過(guò)敏性紫癜患兒組胺H2受體的變化,從臨床的角度探討組胺H2受體拮抗劑對(duì)過(guò)敏性紫癜組胺受體表達(dá)的影響。 4、檢測(cè)過(guò)敏性紫癜患兒急性期、恢復(fù)期及健康兒童血清IL-8水平差異,了解細(xì)胞因子IL-8與過(guò)敏性紫癜的關(guān)系。 5、比較過(guò)敏性紫癜非腎炎組、腎炎組血清IL-8含量差異,了解IL-8是否與過(guò)敏性紫癜腎炎的發(fā)病相關(guān)。 研究方法 1、組胺H2、H4受體在過(guò)敏性紫癜患兒炎癥細(xì)胞中的表達(dá) 選取來(lái)我院就診的兒童,過(guò)敏性紫癜急性期患兒、過(guò)敏性紫癜恢復(fù)期患兒、健康體檢兒童各32例,用實(shí)時(shí)定量熒光聚合酶鏈反應(yīng)(Real-time fluorescent quantification PCR, RT-PCR)及Western蛋白印跡法(Western blot)檢測(cè)三組對(duì)象外周血單個(gè)核細(xì)胞(PBMCs)中H2、H4受體的基因含量及蛋白表達(dá)。 2、過(guò)敏性紫癜急性期患兒予組胺H2受體拮抗劑西咪替丁治療前后H2R的表達(dá) 選取來(lái)我院就診的過(guò)敏性紫癜急性期患兒,分為西咪替丁組、非西咪替丁組,用實(shí)時(shí)定量熒光聚合酶鏈反應(yīng)(Real-time fluorescent quantification PCR, RT-PCR)及Western蛋白印跡法(Western blot)檢測(cè)兩組治療前后H2R的基因含量和蛋白表達(dá)。 3、血清IL-8在過(guò)敏性紫癜中的水平 采用酶聯(lián)免疫吸附試驗(yàn)(emzyme linked immunosorbent asssy, ELISA)檢測(cè)過(guò)敏性紫癜急性期患兒、過(guò)敏性紫癜恢復(fù)期患兒、健康體檢兒童外周血血清IL-8的含量。 結(jié)果 1、組胺H2、H4受體在過(guò)敏性紫癜中的表達(dá) 過(guò)敏性紫癜急性期患兒?jiǎn)蝹�(gè)核細(xì)胞中H2R的mRNA和蛋白的表達(dá)均高于過(guò)敏性紫癜恢復(fù)期兒童及健康兒童(p0.05),過(guò)敏性紫癜恢復(fù)期患兒H2R受體表達(dá)與健康兒童相比無(wú)顯著差異(p0.05)。過(guò)敏性紫癜急性期組H2R的mRNA水平為(1.80±0.49),恢復(fù)期H2R的mRNA為(1.22±0.12),健康對(duì)照組水平為(1.22±0.18),過(guò)敏性紫癜急性期患兒PBMCs中H2R mRNA表達(dá)高于過(guò)敏性紫癜恢復(fù)期患兒、健康兒童(p0.05),過(guò)敏性紫癜恢復(fù)期、健康兒童之間表達(dá)量無(wú)統(tǒng)計(jì)學(xué)差異(p0.05)。 過(guò)敏性紫癜急性期患兒?jiǎn)蝹�(gè)核細(xì)胞中H4R的mRNA和蛋白的表達(dá)均高于過(guò)敏性紫癜恢復(fù)期兒童及健康兒童(p0.05),過(guò)敏性紫癜恢復(fù)期兒童H4受體表達(dá)與健康兒童相比無(wú)顯著差異(p0.05)。過(guò)敏性紫癜急性期H4R的mRNA水平為(4.49±0.99),恢復(fù)期H4R的mRNA為(1.49±0.32),健康對(duì)照組H4R的mRNA水平為(1.44±0.33),過(guò)敏性紫癜急性期患兒PBMCs中H4RmRNA表達(dá)高于過(guò)敏性紫癜恢復(fù)期患兒、健康兒童(p0.05),過(guò)敏性紫癜恢復(fù)期、健康兒童之間表達(dá)量無(wú)統(tǒng)計(jì)學(xué)差異(p0.05) 將過(guò)敏性紫癜急性期患兒根據(jù)是否有血尿、蛋白尿等尿檢異常分為非腎炎組(NHSPN組)和腎炎組(HSPN組)。非腎炎組患兒H2R的mRNA水平為(1.69±0.52),腎炎組的mRNA水平為(1.88±0.47),健康對(duì)照組H2R的mRNA水平為(1.22±0.18)。非腎炎組患兒H4R的mRNA水平為(4.59±1.34),腎炎組的H4R的mRNA水平為(4.39±0.53),健康對(duì)照組H4R的mRNA水平為(1.44±0.33)。非腎炎組、紫癜腎炎組H2R表達(dá)水平均顯著高于健康對(duì)照組(p0.05),非腎炎組和腎炎組兩組間H2R表達(dá)水平無(wú)明顯差異(p0.05)。過(guò)敏性紫癜非腎炎組、腎炎組H4R表達(dá)水平均顯著高于健康對(duì)照組(p0.05),非腎炎組和腎炎組兩組間H4R表達(dá)量無(wú)顯著差異(p0.05)。 2、過(guò)敏性紫癜急性期患兒予組胺H2受體拮抗劑西咪替丁治療前后組胺H2R的表達(dá) 西咪替丁組治療前H2R的mRNA水平為(1.77±0.51),非西咪替丁組治療前H2R的mRNA水平為(1.82±0.50),健康對(duì)照組H2R的mRNA水平為(1.22±0.18),西咪替丁組、非西咪替丁組治療前H2R水平均高于健康對(duì)照組(p0.01)。但西咪替丁組、非西咪替丁組治療前H2R的mRNA表達(dá)無(wú)顯著性差異(p0.05),具有可參照意義。經(jīng)治療,西咪替丁組治療后H2R的mRNA水平為(1.07±0.05),非西咪替丁組治療后H2R的mRNA水平為(1.44±0.34),兩組治療后H2R的mRNA表達(dá)水平有顯著差異(p0.05),西咪替丁治療組H2RmRNA水平變化較大。 3、血清IL-8在過(guò)敏性紫癜中的水平 過(guò)敏性紫癜急性期患兒血清中,IL-8的含量為(332.04±83.48)pg/ml,恢復(fù)期患兒血清中IL-8的含量為(97.75±18.16)pg/ml,健康對(duì)照組患兒血清中IL-8的含量為(90.9±9.73)pg/ml。過(guò)敏性紫癜急性期患兒血清中IL-8水平明顯高于過(guò)敏性紫癜恢復(fù)期患兒、健康兒童(p0.05),過(guò)敏性紫癜恢復(fù)期、健康兒童之間水平量無(wú)統(tǒng)計(jì)學(xué)差異(p0.05)。 過(guò)敏性紫癜非腎炎組患兒血清IL-8的含量為(300.86±98.62)pg/ml,腎炎組血清中IL-8的含量為(359.56±57.24)pg/ml,健康對(duì)照組IL-8水平為(90.9±9.73)pg/ml。腎炎組患兒、非腎炎組患兒血清中IL-8水平均明顯高于健康對(duì)照組患兒IL-8水平。過(guò)敏性紫癜非腎炎組和腎炎組兩組間IL-8含量有明顯差異(p0.05),腎炎組IL-8水平高于非腎炎組。 結(jié)論 1、過(guò)敏性紫癜急性期患兒?jiǎn)蝹�(gè)核細(xì)胞中組胺H2R、H4R蛋白和nRNA的表達(dá)均明顯高于過(guò)敏性紫癜恢復(fù)期患兒、健康兒童。經(jīng)治療癥狀緩解的過(guò)敏性紫癜恢復(fù)期患兒H2R、H4R的表達(dá)能降至健康兒童水平。組胺H2R, H4R可能參與過(guò)敏性紫癜發(fā)病過(guò)程。 2、過(guò)敏性紫癜急性期非腎炎組、腎炎組患兒?jiǎn)蝹�(gè)核細(xì)胞中組胺H2R、H4R蛋白和mRNA的表達(dá)均高于健康兒童,過(guò)敏性紫癜非腎炎組和腎炎組兩組間H2R、H4R表達(dá)水平無(wú)明顯差異。過(guò)敏性紫癜是否累及腎臟,組胺H4R的表達(dá)無(wú)明顯差異。 3、過(guò)敏性紫癜急性期患兒,予H2R受體拮抗劑西咪替丁治療組治療前后組胺H2R變化較非西咪替丁治療組組胺H2R變化有明顯差異,西咪替丁可減少組胺受體的表達(dá)。 4、過(guò)敏性紫癜急性期患兒血清IL-8含量明顯高于過(guò)敏性紫癜恢復(fù)期患兒、健康兒童。經(jīng)治療癥狀緩解的過(guò)敏性紫癜恢復(fù)期患兒IL-8含量能降至健康兒童水平。IL-8可能參與過(guò)敏性紫癜發(fā)病過(guò)程。 5、過(guò)敏性紫癜腎炎患兒血清IL-8含量高于非腎炎組,IL-8可能參與紫癜性腎炎的發(fā)生及發(fā)展。
[Abstract]:research objective
1, to compare the expression of histamine H2 and H4 receptor on the surface of peripheral blood mononuclear cells in children with anaphylactoid purpura during the acute period, recovery and health examination, and to explore the relationship between histamine H2, H4 receptor and Henoch Schonlein purpura.
2, compare the difference of histamine H2 and H4 receptor expression between Henoch Schonlein purpura nephritis group and Henoch Schonlein purpura nephritis group, and understand the correlation between histamine receptor and Henoch Schonlein purpura nephritis.
3, the changes of histamine H2 receptor in children with Henoch Schonlein purpura before and after cimetidine treated with histamine H2 receptor antagonist were compared, and the effect of histamine H2 receptor antagonist on the expression of histamine receptor in Henoch Schonlein purpura was investigated from a clinical point of view.
4, to detect the difference of serum IL-8 levels between children with acute purpura in acute stage, convalescent stage and healthy children, and to understand the relationship between cytokine IL-8 and allergic purpura.
5, to compare the difference of serum IL-8 content between the nephritis group and the non nephritis group of Henoch Schonlein purpura, and to know whether IL-8 is related to the pathogenesis of Henoch Schonlein purpura nephritis.
research method
1, the expression of histamine H2 and H4 receptors in inflammatory cells of children with Henoch Schonlein purpura.
Children in our hospital, children with acute Henoch Schonlein purpura, children with anaphylactoid purpura, 32 children in healthy physical examination were selected, and three groups of peripheral blood mononuclear cells (PBMCs) were detected by real-time quantitative fluorescent polymerase chain reaction (Real-time fluorescent quantification PCR, RT-PCR) and Western Western blot (Western blot). Gene content and protein expression of H2, H4 receptor.
2, the expression of H2R in the acute phase of Henoch Schonlein purpura before and after treatment with histamine H2 receptor antagonist cimetidine.
Children with anaphylactoid purpura in our hospital were selected to be divided into cimetidine group, non cimetidine group, Real-time fluorescent quantification PCR (RT-PCR) and Western Western blot (Western blot) to detect the gene content and protein expression of H2R in two groups before and after treatment.
3, the level of serum IL-8 in Henoch Schonlein purpura
Emzyme linked immunosorbent asssy (ELISA) was used to detect the levels of serum IL-8 in children with anaphylactoid purpura at the acute stage of anaphylactoid purpura, in children with anaphylactoid purpura, and in healthy children.
Result
1, the expression of histamine H2, H4 receptor in Henoch Schonlein purpura
The expression of mRNA and protein of H2R in mononuclear cells in children with anaphylactoid purpura was higher than that of children with anaphylactoid purpura and healthy children (P0.05). There was no significant difference in the expression of H2R receptor in children with Henoch Schonlein purpura at the recovery period (P0.05). The mRNA level of H2R in anaphylactoid purpura acute phase group was (1.80 + 0.49), and the recovery period H2R The mRNA was (1.22 + 0.12), and the level of the healthy control group was (1.22 + 0.18). The expression of H2R mRNA in the children with anaphylactoid purpura at the acute stage of PBMCs was higher than that of the anaphylactoid purpura. There was no statistical difference between healthy children (P0.05) and the recovery period of anaphylactoid purpura. There was no statistical difference between the healthy children (P0.05).
The expression of mRNA and protein of H4R in mononuclear cells in children with anaphylactoid purpura was higher than that of children with anaphylactoid purpura and healthy children (P0.05). There was no significant difference in the expression of H4 receptor in children with anaphylactoid purpura at the recovery period (P0.05). The mRNA level of H4R in anaphylactoid purpura was (4.49 + 0.99) and H4R mR in the recovery period. NA was (1.49 + 0.32), and the level of mRNA of H4R in healthy control group was (1.44 + 0.33). The expression of H4RmRNA in children with anaphylactoid purpura at the acute stage of PBMCs was higher than that of children with anaphylactoid purpura, healthy children (P0.05), anaphylactoid purpura recovery period, and no difference in expression between healthy children (P0.05).
The patients with anaphylactoid purpura at acute stage were divided into non nephritis group (group NHSPN) and nephritis group (group HSPN). The mRNA level of H2R in the non nephritis group was (1.69 + 0.52), the mRNA level of the nephritis group was (1.88 + 0.47), and the mRNA level of H2R in the healthy control group was (1.22 + 0.18). The mRNA level of H4R in the non nephritis group was in the control group. The mRNA level of H4R in nephritis group was (4.39 + 0.53), and the mRNA level of H4R in the healthy control group was (1.44 + 0.33). The H2R expression level of the non nephritis group and the purpura nephritis group was significantly higher than that of the healthy control group (P0.05). There was no significant difference in the H2R expression level between the non nephritis group and the glomerulonephritis group (P0.05). The allergic purpura non nephritis group and the nephritis group were H4R (H4R). The level of expression was significantly higher than that of healthy controls (P0.05). There was no significant difference in H4R expression between the two groups of non nephritis and nephritis (P0.05).
2, the expression of histamine H2R before and after treatment with histamine H2 Receptor Antagonist Cimetidine in children with acute Henoch Schonlein purpura.
The mRNA level of H2R before treatment in cimetidine group was (1.77 + 0.51). The mRNA level of H2R before treatment in non cimetidine group was (1.82 + 0.50), mRNA level of H2R in healthy control group was (1.22 + 0.18). The level of H2R in cimetidine group and non cimetidine group before treatment was higher than that of healthy control group (P0.01). But cimetidine group and non cimetidine group treated H2R m before treatment. There was no significant difference in RNA expression (P0.05). The mRNA level of H2R in cimetidine group was (1.07 + 0.05) after treatment. The mRNA level of H2R after treatment in non cimetidine group was (1.44 + 0.34). The mRNA expression level of H2R after treatment in two groups was significantly different (P0.05), and the H2RmRNA level of cimetidine treatment group changed greatly.
3, the level of serum IL-8 in Henoch Schonlein purpura
In the serum of children with anaphylactoid purpura, the content of IL-8 was (332.04 + 83.48) pg/ml, and the content of IL-8 in the serum of the children with the recovery period was (97.75 + 18.16) pg/ml. The serum IL-8 content in the healthy control group was (90.9 + 9.73) pg/ml. allergic purpura. The serum IL-8 level was significantly higher than that of the children with the anaphylactoid purpura. In children (P0.05), there was no significant difference in the level of children with Henoch Schonlein purpura at the recovery stage (P0.05).
The serum IL-8 content of the children with anaphylactoid purpura non nephritis group was (300.86 + 98.62) pg/ml, the content of IL-8 in the serum of the nephritis group was (359.56 + 57.24) pg/ml, and the level of IL-8 in the healthy control group was (90.9 + 9.73) pg/ml. nephritis group. The serum IL-8 level of the children in the non nephritis group was significantly higher than that of the healthy control group. The non kidney of the allergic purpura was not the kidney. There was significant difference in IL-8 content between the two groups of inflammation group and nephritis group (P0.05), and IL-8 level in nephritis group was higher than that in non nephritis group.
conclusion
1, the expression of histamine H2R, H4R protein and nRNA in the mononuclear cells of children with anaphylactoid purpura was significantly higher than that of the children with anaphylactoid purpura, and healthy children. The expression of H4R could be reduced to the level of healthy children after the treatment of symptomatic anaphylactoid purpura, and the expression of H4R could be reduced to the level of healthy children. Histamine H2R, H4R may be involved in the pathogenesis of anaphylactoid purpura.
2, in the acute phase of anaphylactoid purpura non nephritis group, the expression of histamine H2R, H4R protein and mRNA in the mononuclear cells of the nephritis group were all higher than that of the healthy children. There was no significant difference in the expression of H4R between the two groups of the anaphylactoid purpura non nephritis group and the nephritis group. There was no significant difference in the expression of the renal and histamine H4R in the Henoch Schonlein purpura.
3, in children with anaphylactoid purpura at acute stage, the changes of histamine H2R in the group of H2R receptor antagonist cimetidine treatment group were significantly different from those in the non cimetidine group before and after treatment. The expression of histamine receptor was reduced by cimetil Ding Ke.
4, the serum levels of IL-8 in children with anaphylactoid purpura were significantly higher than those in the recovery period of anaphylactoid purpura, and in healthy children, the content of IL-8 in the children with anaphylactoid purpura during the treatment of symptomatic remission could be reduced to the level of.IL-8 in healthy children and may be involved in the pathogenesis of anaphylactoid purpura.
5, the serum IL-8 level of children with Henoch Schonlein purpura nephritis is higher than that of non nephritis group, and IL-8 may be involved in the occurrence and development of purpuric nephritis.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R725.5

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