ABSTRACT
Objective:To investigate the effects of pregnancy and lactation nonylphenol (NP) exposure on the balance of Treg/Th17 cells in the brain of offspring mice and the related mechanisms.Methods:Thirty pregnant C57BL/6 mice were randomly divided into three groups: control group (drinking distilled water), and NP-treated groups (drinking 0.2 μg/ml or 2.0 μg/ml NP water solution). ELISA kit was used to analyze the levels of TNF-α, IFN-γ and IL-17, flow cytometry was used to analyze the frequency of Treg and Th17 cells in spleen, quantitative RT-PCR was used to analyze the RORγt, Foxp3 mRNA, Western blot was used to analyze the protein expression of RORγt, Foxp3 and PI3K/Akt/mTOR signal pathway, and immunofluorescence was used to analyze the expression of Iba1 in the brain tissue of offspring mice.Results:Compared with the control group, NP exposure increased the serum levels of IL-17 and TNF-α in male offspring mice ( P<0.05), and decreased the levels of IFN-γ( P<0.05). Flow cytometry analysis showed that the percentage of Th17 cells in the spleen of male offspring mice exposed to NP (0.2 μg/ml or 2.0 μg/ml) was significantly higher than that of the control group, while the percentage of Tregs cells was lower. Compared with the control group, the expression levels of Foxp3 proteins in the brain tissue of male offspring mice exposed to NP (0.2 μg/ml or 2.0 μg/ml) was significantly lower, accompanied by a dramatic increase in RORγt protein levels ( P<0.05). Similar mRNA expression was also observed in qRT-PCR analysis. The protein expression levels of mTOR (p-mTOR) and its upstream related regulators[PI3K, p-Akt (Ser473), p-Akt (Thr308)] in the brain of male offspring mice increased gradually during the period of exposure to NP( P<0.05). Immunofluorescence analysis showed that compared with the control group, the number of Iba1 positive cells in brain tissue of male offspring mice exposed to NP (0.2 μg/ml or 2.0 μg/ml) increased significantly ( P<0.05). Conclusions:Maternal exposure to NP during pregnancy and lactation may affect the development/function of neurons in offspring through neuroimmune axis, and increase the risk of neurodevelopmental disorders in offspring.
ABSTRACT
Objective To observe the molecular mechanism of adaptive response of the kidney and skeleton and brain issues in the high altitude hypoxia; To discuss the unity of yin and yang oscillation relationship of kidney and brain marrow.Methods SPF KM mice were randomly divided into control group and model group according to random number table method. Mice in the model group were exposed to high altitude hypoxia cabin for successive 21 d. On the 22nd day, mice got out of the cabin and their body weight was measured, and then they were put to death through eyeball blood sampling. The activities of lactic LDH and Na+-K+-ATPase in brain tissue were detected by spectrophotometric colorimetry. The PFK activities of brain and skeletal muscle were detected by ELISA. Meanwhile the contents of EPO and EPOR in the kidney were measured by ELISA. The mRNA expressions of HIF-1α and AQP-4 in brain were assessed by RT-PCR. At the same time, the protein expressions of HIF-1α and AQP-1 in brain and the protein expression of Mb in skeletal muscle were detected by Western blot.Results Compared with the normal group, the LDH and PFK in brain tissue and the content of EPO in kidney tissue were all raised in the model group(P<0.05). Meanwhile the mRNA expressions of HIF-1α and AQP-4 and the protein expressions of HIF-1α and AQP-1 in brain were all increased in the mice from the model group; the activities of PFK and the protein expression of Mb in skeletal muscle were also raised in the model group. But the activity of Na+-K+-ATPase in brain tissue and the content of EPOR in kidney tissue both decreased in the model group (P<0.05,P<0.01).Conclusion Adaptive response and the unity of yin and yang oscillation relationship between kidney, skeleton and brain tissue happen in high altitude hypoxia.