1.2.4 統(tǒng)計學(xué)處理 觀察同一患者鼻息肉組織及下鼻甲黏膜成纖維細胞中各型組胺受體的表達,用實時擴增了解1型及2型組胺受體表達程度.將目的基因交點與βactin基因交點比值作為受體的表達程度并以均數(shù)±標準偏差(±SD)來表示.用SPSS12.0統(tǒng)計軟件對數(shù)據(jù)進行統(tǒng)計學(xué)處理,進行MannWhiteny’s U檢驗.
2 結(jié)果
RTPCR結(jié)果表明,可見1型及2型組胺受體mRNA表達,未見3型及4型組胺受體表達(圖2).實時擴增結(jié)果見,鼻息肉組織成纖維細胞內(nèi)1型及2型組胺受體mRNA的表達較下鼻甲鼻黏膜成纖維細胞低,但相比較均無統(tǒng)計學(xué)意義(P=0.19,P=0.14,圖3).流式細胞分析實驗結(jié)果示,無刺激時1型組胺受體蛋白的表達量在鼻息肉組織及下鼻甲黏膜組織中均極低,無顯著性差異(圖4),2型組胺受體蛋白的表達在下鼻甲黏膜組織較多(圖5).
3 討論
組胺生成于嗜中性粒細胞及肥大細胞,參與各種過敏反應(yīng).氣道上皮微環(huán)境中的肥大細胞可合成組胺,增加氣道的滲透性[15].Jordana等[16]認為,組胺可使體外培養(yǎng)的成纖維細胞增殖,其作用是通過2型組胺受體完成的;thymikine可使體外培養(yǎng)的人皮膚成纖維細胞增殖,而組胺可通過1型組胺受體促進蛋白激酶C及佛波酯酵素活性,抑制thymikine對成纖維細胞的增殖作用[17].目前,對在鼻息肉的形成過程中成纖維細胞及成纖維細胞中的組胺受體所起的作用雖然尚未完全闡明,但可以肯定的是鼻息肉的形成僅與1型及2型組胺受體有關(guān),而與3型及4型組胺受體則無關(guān).本研究結(jié)果亦表明,鼻息肉成纖維細胞中只可見1型及2型組胺受體的表達,而未見3型及4型的表達.目前雖然對鼻息肉的形成過程尚無統(tǒng)一的認識,但與變應(yīng)性鼻炎有關(guān)系的假設(shè)已被受重視.Okayama等[18]認為,人鼻黏膜組織中分布的1型組胺受體主要分布于小動脈、靜脈及海棉靜脈竇等血管內(nèi)皮細胞內(nèi),而血管平滑肌及鼻黏膜下腺體等組織中則未見分布.Boucher等[19]報道,鼻黏膜切片組織中存在2型組胺受體的表達,且在變應(yīng)性鼻炎鼻黏膜組織中的表達強度明顯高于非變應(yīng)性鼻炎,且原位雜交分析亦支持上述結(jié)果.本研究采用流式細胞分析方法觀察了組胺受體蛋白的表達,發(fā)現(xiàn)其表達量較少,但組胺受體蛋白可以細胞內(nèi)及外的形態(tài)表達,而流式細胞分析方法卻不能準確地測定細胞內(nèi)的蛋白量.蛋白表達較少的原因考慮可能與如下幾點相關(guān);1)可以肯定鼻息肉與下鼻甲黏膜成纖維細胞中有大量的組胺受體mRNA表達;2)組胺受體蛋白合成后受到某些其他代謝產(chǎn)物的影響,迅速溶解;3)大部分組胺受體蛋白僅在細胞內(nèi)停留,而未到達細胞膜表面;4)可能在某些因素的影響下蛋白質(zhì)翻譯受阻礙;5)需要特定的條件,如各種細胞因子刺激或特殊酶參與等.
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