摘要
目的建立小鼠妊娠期大气细颗粒物(fine particulate matter,AD≤2.5um,PM_(2.5))暴露模型,探讨妊娠期PM_(2.5)暴露对新生小鼠肝脏发育的影响。方法孕鼠随机分为对照组、PM_(2.5)低剂量组、PM_(2.5)中剂量组,利用气管滴注方法分别向PM_(2.5)低、中剂量组注入浓度为0.2592μg/μl、1.56695μg/μl的30μl PM_(2.5)混悬液,对照组注入相同体积磷酸盐缓冲液。孕期间断滴注7次。HE染色观察新生小鼠肝脏的组织结构变化,免疫组织化学技术和Westem blot技术观察新生小鼠肝脏PCNA和caspase-3蛋白的表达情况。结果 PM_(2.5)中剂量组新生小鼠肝脏中可见明显病理学改变,肝细胞水肿,明显脂肪变性。随着滴注剂量的增加,免疫组织化学技术显示肝脏PCNA阳性细胞数逐渐减少,easpase-3阳性细胞数逐渐增多;Western blot技术显示PM_(2.5)暴露后新生小鼠肝脏PCNA蛋白表达量有不同程度降低,caspase-3蛋白表达量升高。结论妊娠期PM_(2.5)暴露可通过抑制肝细胞的增殖和促进其凋亡,从而影响新生小鼠肝脏的发育,并产生病理损害。
Objective To establish mice model of exposure to atmospheric fine particles matter(AD ≤2.5um,PM_(2.5)) during gestation to investigate the effects of PM_(2.5) on the development of liver in newborns.Methods Pregnant mice were randomly divided into three groups:the control group,PM_(2.5) low dose group,and PM_(2.5) medium dose group.The tracheal instillation method was used to inject the experimental groups of mice with 30μl PM_(2.5) suspension of different doses(low dose:0.2592 μg/μl,medium dose:1.56695 μg/μl) respectively,and the control group with phosphate buffer saline of the same volume.The instillation was intermittently performed for a total of seven times during pregnancy.The liver structural change was observed with HE staining in newborn mouse,and the expressions of PCNA and caspase-3 proteins were detected by immunohistochemistry and Western blot.Results Compared with the control group,obvious pathological changes were observed in the livers of newborn mice in the medium dose group of PM_(2.5) exposure,including liver cell edema and significant fatty degeneration.Immunochemistry results showed that the number of PCNA positive liver cells decreased with the increase of instilled PM_(2.5) dose,while that of caspase-3 positive cells increased.Western blot showed that the expression of PCNA protein decreased while caspase-3 increased to varying degrees in the newborn mouse liver after gestational PM_(2.5) exposure.Conclusion PM_(2.5) exposure during gestation can affect newborn mouse liver development and cause pathological lesion through the inhibition of liver cell proliferation and promotion of its apoptosis.
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2016年第5期406-410,共5页
Chinese Journal of Histochemistry and Cytochemistry
基金
山东省自然科学基金项目(ZR2014JL016)
