體外循環(huán)手術(shù)對紅細(xì)胞CD35、CD59及T淋巴細(xì)胞免疫的影響

1、體外循環(huán)手術(shù)對紅細(xì)胞CD35、CD59及T淋巴細(xì)胞免疫的影響 10-01-29 13:28:00 編輯：studa20 作者：胡金川,田亞平,李佳春,董培青,萬彩紅,谷 峰,駱 藎,高艷紅,江朝光【摘要】 目的 探討體外循環(huán)(extracorporeal circulation,ECC)手術(shù)對紅細(xì)胞CD35、CD59及T淋巴細(xì)胞免疫功能的影響。 方法 選擇ECC手術(shù)病例18例，于轉(zhuǎn)機(jī)前、體溫最低點(diǎn)、轉(zhuǎn)機(jī)末和術(shù)后1 d、3 d、7 d晨采集患者靜脈血，測定紅細(xì)胞CD35、CD59，以及T淋巴細(xì)胞CD3、CD4和CD8。 結(jié)果 ECC前至ECC末，CD35+紅細(xì)胞(CD35+-E)百分率、CD59

2、幾何平均熒光強(qiáng)度比值(CD59-GMFIR)和CD35-GMFIR均逐漸降低，前二者在術(shù)后1 d至7 d逐漸回升，而CD35-GMFIR未見明顯恢復(fù)趨勢。ECC末、術(shù)后1 d時CD35+-E百分率和CD35-GMFIR，以及體溫最低點(diǎn)、ECC末和術(shù)后1 d時CD59-GMFIR均顯著低于ECC前(P0.01或P0.05)。至術(shù)后7 d時，CD35+-E百分率、CD35-GMFIR和CD59-GMFIR仍未恢復(fù)至ECC前水平。從ECC前至術(shù)后7 d，CD3+ T淋巴細(xì)胞(CD3+-T)百分率、CD3+CD4+-T百分率和CD4+-T/CD8+-T比值均呈現(xiàn)先降低后升高的趨勢，體溫最低點(diǎn)、ECC末

3、、術(shù)后1 d和術(shù)后3 d的CD3+-T百分率、CD3+CD4+-T百分率均顯著低于ECC前(P0.01或P0.05)，至術(shù)后7 d恢復(fù)至接近于ECC前水平，而CD4+-T/CD8+-T比值則恢復(fù)至超過ECC前水平。 結(jié)論 ECC手術(shù)使紅細(xì)胞CD35、CD59及T淋巴細(xì)胞免疫功能下降，后者于術(shù)后7 d恢復(fù)，前者恢復(fù)期長于后者。 【關(guān)鍵詞】 體外循環(huán)；紅細(xì)胞免疫；CD35抗原；CD59抗原；T淋巴細(xì)胞CD59 of erythrocytes, and T lymphocytes immune. METHODS Eighteen cases of heart operation under ECC

4、were included in this study. Venous blood samples were collected at pre-ECC, time of minimum body temperature, end of ECC, and 1 day, 3 days, 7 days after operation, then CD35 and CD59 on erythrocytes, and CD3, CD4 and CD8 on T lymphocytes were measured by flow cytometry immediately. RESULTS The CD3

5、5 positive erythrocytes(CD35+-E) percentage, geometric mean fluorescence intensity ratio of CD59(CD59-GMFIR) and CD35-GMFIR at time of minimum body temperature and end of ECC decreased, and the former two turned to recover to the level at pre-ECC, while CD35-GMFIR didnt have the same trend. CD35+-E

6、percentage, CD35-GMFIR at end of ECC and 1 day after operation, and also CD59-GMFIR at time of minimum body temperature, end of ECC and 1 day after operation were lower than those at pre-ECC(P0.01 or P0.05), and all the indexes didnt return to the level at pre-ECC until 7 days after operation. The p

7、ercentages of CD3 positive T lymphocytes(CD3+-T) and CD3+CD4+-T, and the ratio of CD4+-T/CD8+-T showed similar tendency as the indexes of red-cell immune from pre-ECC to 7 days after operation. The percentages of CD3+-T and CD3+CD4+-T at time of minimum body temperature, end of ECC and 1 day, 3 days

8、 after operation were all lower than those at pre-ECC (P0.01 or P0.05), and returned to approach the pre-ECC level at 7 days after operation, while CD4+-T/CD8+-T ratio exceeded the pre-ECC level at that time. CONCLUSION CD35 and CD59 on erythrocytes, and T lymphocytes immune function decrease after

9、ECC operation, the latter can recover at 7 days after operation, and the recovery period of the former is longer than that of the latter. Key words： Extracorporeal circulation; Red-cell immune; CD35 antigen; CD59 antigen; T lymphocytes 體外循環(huán)(extracorporeal circulation，ECC)是心內(nèi)直視手術(shù)治療心臟疾病的一種重要手段。由于其是一非生

10、理性循環(huán)，在ECC過程中，管路和氧合器材料、機(jī)械作用、氧合方式、溫度應(yīng)激等對血液成分的損害而引起炎性反應(yīng)和免疫抑制是導(dǎo)致并發(fā)癥的主要原因。紅細(xì)胞占血液有形成分的99%以上，承擔(dān)著呼吸和免疫兩種功能，其免疫黏附能力占血細(xì)胞的85%以上1，在血液免疫中起重要作用。紅細(xì)胞在ECC中受損最為嚴(yán)重，研究ECC對紅細(xì)胞免疫分子及T淋巴細(xì)胞免疫的影響，比較二者變化趨勢的異同，有助于探討ECC導(dǎo)致免疫抑制的機(jī)制，為有效防治提供依據(jù)。 1 資料與方法 1.1 臨床資料 心血管外科ECC手術(shù)病例18例，男10例，女8例，年齡1971(45.616.2)歲?；颊咝g(shù)前未使用過免疫抑制劑，無感染、腫瘤及自身免疫性疾病等

11、致免疫低下的合并癥。 1.2 手術(shù)方法 采用靜脈復(fù)合氣管內(nèi)全麻，常規(guī)正中開胸插管建立ECC。體內(nèi)肝素化(3 mg/kg)。ECC采用膜式氧合器，淺低溫，經(jīng)升主動脈根部順行灌注高鉀冷血停搏液保護(hù)心肌，心臟停跳下行心內(nèi)直視手術(shù)。ECC時間69114(83.218.6)min，升主動脈阻斷4668(58.28.6)min。停止ECC后常規(guī)魚精蛋白中和肝素。術(shù)畢患者入重癥監(jiān)護(hù)室。 1.3 實(shí)驗(yàn)方法 于ECC前、體溫最低點(diǎn)、ECC末及術(shù)后1 d、3 d、7 d分別采集患者靜脈血于EDTA抗凝管中。 紅細(xì)胞CD35和CD59測定：EDTA抗凝血離心后，取5105個紅細(xì)胞/管，設(shè)同型對照管和測試管，分別加相應(yīng)熒光標(biāo)記抗體7 l，室溫避光30 min，PBS洗滌后重懸，用美國Becton Dickinson公司FACS Calibur流式細(xì)胞儀測試。計(jì)算CD35陽性紅細(xì)胞(CD35+-E)百分率和單個紅細(xì)胞CD35及CD59幾何平均熒光強(qiáng)度比值(geometric mean fluorescence intensity ratio，GMFIR)(測試管GMFI/同型對照管GMFI)。 T淋巴細(xì)胞CD3、CD4和CD8測定：取100 l EDTA抗凝全血，加5 l CD45-FITC/CD4-PE/CD8-ECD/CD3-PC5混合標(biāo)記抗