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In utero hypoxia attenuated acetylcholine-mediated vasodilatation via CHRM3/p-NOS3 in fetal sheep MCA: role of ROS/ERK1/2

Hypertension Research volume 45, pages 1168–1182 (2022)Cite this article

Abstract

Hypoxia can lead to adult middle cerebral artery (MCA) dysfunction and increase the risk of cerebrovascular diseases. It is largely unknown whether intrauterine hypoxia affects fetal MCA vasodilatation. This study investigated the effects and mechanisms of intrauterine hypoxia on fetal MCA vasodilatation. Near-term fetal sheep were exposed to intrauterine hypoxia. Human umbilical vein endothelial cells (HUVECs) were exposed to hypoxia in cellular experiments. Vascular tone measurement, molecular analysis, and transmission electron microscope (TEM) were utilized to determine vascular functions, tissue anatomy, and molecular pathways in fetal MCA. In fetal MCA, acetylcholine (ACh) induced reliable relaxation, which was markedly attenuated by intrauterine hypoxia. Atropine, P-F-HHSiD, L-NAME, and u0126 blocked most ACh-mediated dilation, while AF-DX 116 and tropicamide partially inhibited the dilation. Indomethacin and SB203580 did not significantly change ACh-mediated dilation. Tempol and PS-341 could restore the attenuated ACh-mediated vasodilatation following intrauterine hypoxia. The mRNA expression levels of CHRM2 and CHRM3 and the protein levels of CHRM3, p-NOS3, SOD2, ERK1/2, p-ERK1/2, MAPK14, and p-MAPK14 were significantly reduced by intrauterine hypoxia. The dihydroethidium assay showed that the production of ROS was increased under intrauterine hypoxia. TEM analysis revealed endothelial cells damaged by intrauterine hypoxia. In HUVECs, hypoxia increased ROS formation and decreased the expression of CHRM3, p-NOS3, SOD1, SOD2, SOD3, ERK1/2, p-ERK1/2, and p-MAPK14, while tempol and PS-341 potentiated p-NOS3 protein expression. In conclusion, in utero hypoxia reduced ACh-mediated vasodilatation in ovine MCA predominantly via decreased CHRM3 and p-NOS3, and the decreased NOS3 bioactivities might be attributed to ROS and ERK1/2.

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Acknowledgements

ZX designed the experiments. JT and NL performed surgery and experiments. PZ performed anesthesia. YH and HS performed vascular measurements and molecular tests. YZ (YZ, YZ, YZ) and YY helped with the molecular measurements and cell culture. YH and JT wrote the manuscript, and ZX revised it.

Funding

This work was supported by grants 2019YFA0802601, NSFC (81771592, 82101761), Natural Science Foundation of Jiangsu Province (BK20200194), Suzhou Natural Science Foundation (SYS2019042, KJXW2019006), Wuxi Taihurencai Project Fund, and Postgraduate Research & Practice Innovation Program of Jiangsu Province (KYCX20-2693).

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  1. These authors contributed equally: Yun He, Hongyu Su

Authors and Affiliations

  1. Institute for Fetology, First Hospital of Soochow University, Suzhou, China

    Yun He, Hongyu Su, Yingying Zhang, Pengjie Zhang, Yumeng Zhang, Jiaqi Tang & Zhice Xu

  2. Perenatology Laboratory, Maternal and Child Health Care Hospital of Wuxi, Wuxi, China

    Na Li, Yingying Zhang, Yang Ye & Zhice Xu

  3. Dushu Lake Hospital Affiliated to Soochow University, Suzhou, China

    Yueming Zhang

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He, Y., Su, H., Li, N. et al. In utero hypoxia attenuated acetylcholine-mediated vasodilatation via CHRM3/p-NOS3 in fetal sheep MCA: role of ROS/ERK1/2. Hypertens Res 45, 1168–1182 (2022). https://doi.org/10.1038/s41440-022-00935-y

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