连续性血液净化对肺泡上皮细胞Connexin43的影响  

Effects of continuous blood purification on Connexin43 in human alveolar epithelial cells

作　　者：杨溢[1] 甘华[1] 李正荣[1] 文以君[1] 王喜超[1] 

机构地区：[1]重庆医科大学附属第一医院肾内科,重庆400016

出　　处：《第四军医大学学报》2009年第13期1219-1222,共4页Journal of the Fourth Military Medical University

摘　　要：目的:研究连续性血液净化(CBP)对重症急性胰腺炎(SAP)伴急性肺损伤(ALI)患者血清诱导的人肺泡上皮细胞(AECs)间隙连接蛋白Connexin43(Cx43)表达的影响.方法:采集健康志愿者清晨空腹及SAP伴ALI患者CBP治疗前、治疗6h、治疗20h时的血清,采用免疫荧光法、逆转录聚合酶链反应(RT-PCR)半定量法,检测各组血清培养48h后的AECs,Cx43及其mRNA的表达.双抗夹心酶联免疫吸附法(ELISA)检测各组血清肿瘤坏死因子-α(TNF-α)水平.结果:Cx43免疫荧光阳性染色实验各组明显少于健康对照组,且随着CBP治疗时间的延长,阳性染色逐渐增加.Cx43 mRNA的相对表达量在治疗前组(0.08±0.01)较健康对照组(0.57±0.02)显著减少(P<0.01);随治疗时间的延长,治疗6h组(0.23±0.02),治疗20h组(0.36±0.02)其表达逐渐增加(P<0.01).血清TNF-α水平在治疗前组(59.43±4.50)ng/L较健康对照组(16.06±3.68)ng/L显著升高(P<0.01);随治疗时间的延长,治疗6h组(41.16±3.49)ng/L,治疗20h组(34.65±3.22)ng/L,其水平逐渐降低(P<0.01).Cx43 mRNA的表达与TNF-α水平呈负相关.结论:AECs Cx43的减少参与了SAP继发ALI的过程,CBP治疗可通过清除血清TNF-α从而上调Cx43的表达,对呼吸功能起到一定保护作用.AIM： To investigate the effect of continuous blood purification （CBP） on the expression of Connexin43 （ Cx43 ） in human alveolar epithelial ceils （AECs） induced by the serum of patients with severe acute pancreatitis （SAP） accompanied with acute lung injury（ALI）. METHODS ： Fasting serum of healthy volunteers and serum of patients with SAP and ALl at pre-CBP, 6-hour-CBP and 20-hour-CBP were collected. Immunofluorescence staining and RT-PCR were used to detect the expression of Cx43 and Cx43 mRNA in AECs separately cultured by the serum from different groups for 48 h in vitro. ELISA was used to detect the level of serum TNF-α in each group. RESULTS ： Cx43 positive staining in each patient group was significantly less than that in healthy control group. With the increase of the treatment time, the positive staining gradually increased. The relative expression of Cx43 mRNA in pre-CBP group（0.08 ±0.01 ） was significantly less than that in healthy control group（0.57 -0.02） （P 〈0.01 ）. With the increase of the treatment time, the expression gradually increased in 6-hour-CBP group （0.23 ± 0.02 ） and 20-hour-CBP group（0.36± 0.02 ）（ P 〈 0. 01 ）. The level of serum TNF-α in pre-CBP group （ 59.43±4.50 ） ng/L was significantly higher than that in healthy control group （ 16.06 ± 3.68 ） ng/L （ P 〈0. 01 ）. With the increase of the treatment time, the level gradually decreased in 6-hour-CBP group （41. 16 ± 3. 49） ng/L and 20-hour-CBP group（34.65±3. 22） ng/L（P 〈0.01）. The expression of Cx43 mRNA was negatively correlated with the level of TNF-α. CONCLUSION ： The decrease of Cx43 in AECs plays a part in the process of SAP patients with secondary ALl. CBP up-regulates the expression of Cx43 by removing TNF-α and thus protects the respiratory function.

关 键 词：间隙连接蛋白43 急性肺损伤 血液滤过/方法 重症急性胰腺炎 肺泡上皮细胞 

分 类 号：R459.5[医药卫生—治疗学]