梗阻性黄疸患者血清对人肺微血管内皮细胞肌样分化的影响

Effect of serum of patients with obstructive jaundice on myogenic differentiation of human pulmona- ry microvascular endothelial cells

目的评价梗阻性黄疸患者血清对人肺微血管内皮细胞（PMVECs）肌样分化的影响。 方法分离人PMVECs，并传代培养，将培养的人PMVECs分别采用梗阻性黄疸患者血清和健康志愿者血清孵育，于孵育24、48和72 h（T1～3）时在倒置显微镜下观察原代PMVECs形态，采用Western blot法检测PMVECs肌性蛋白（α-SMA、SM-MHC、capolnin）的表达。 结果健康志愿者血清孵育PMVECs calponin、α-SMA蛋白表达阴性，出现少量SM-MHC表达，梗阻性黄疸患者血清孵育PMVECs calponin、α-SMA和SM-MHC表达阳性。与健康志愿者血清比较，梗阻性患者黄疸血清孵育PMVECs SM-MHC表达上调（P〈0.05）；与T1时比较，T2，3时梗阻性黄疸血清孵育PMVECs calponin、α-SMA和SM-MHC表达上调（P〈0.05）；与T2时比较，T3时梗阻性黄疸患者血清孵育PMVECs calponin、α-SMA和SM-MHC表达上调（P〈0.05）。 结论梗阻性黄疸患者血清促进PMVECs肌样分化，可能是肺微血管扩张机制之一。

Objective To evaluale the effects of the serun] of patients with obslruetive jaundice on myogenic differentiation of human pulmonary mierovaseular endothelial cells （PMVECs）. Methods iluman PMVECs were isolaled and then suheultm＇ed. The cultured PMVI＇]Cs were ineuhaled with the serum uf patienls with obstruelive jaundice or with the serum ofheahhy vnlunteers. AI 24, 48 and 72 h of incubation （T2-3）, Ihe inverted microscope was used to observe the morphology of primary PMVECs. The expression of muscular proteins （alpha-smooth muscle actin [ α-SMA], smooth muscle-mysion heavy chain [ SM- MHC] , eapolnin） in PMVECs was detected using Western blot analysis. Results The expression of eal- ponin and a-SMA was negative, and a few SM-MTIC proteins were expressed when PMVECs were ineuimled with the serum of healthy volunteers; the expression ofcalponin, α-SMA and SM-MHC was posiliw when PMVECs were incubated with the serum of patients with obstructive jaundice. Compared with the serum of heahhy volunteers, the expression of SM-MHC was signifieantly up-regulated when PMVECs were incubated with the serum of patients with obstructive jaundice （P〈0.05）. The expression of ealponin, α-SMA and SM-MHC was significantly up-regulated at T2.3 compared with that at T1 , and at T3 compared with that at T2 when PMVECs were incubated with the serum of patients with obstructive jaundice （P〈0.05）. Conclusion The serum of patients with obstructive jaundice promotes myogenic differentiation of human PMVECs, which is probably one of the mechanisms underlying intrapuhnonary mierovascular dilatation.