Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Apr;25(7):3437-3448.
doi: 10.1111/jcmm.16423. Epub 2021 Mar 6.

Hydrogen sulphide reduces hyperhomocysteinaemia-induced endothelial ER stress by sulfhydrating protein disulphide isomerase to attenuate atherosclerosis

Shan Jiang  1 , Wenjing Xu  2 , Zhenzhen Chen  3 , Changting Cui  3 , Xiaofang Fan  1 , Jun Cai  3 , Yongsheng Gong  1 , Bin Geng  1   3
Affiliations

Hydrogen sulphide reduces hyperhomocysteinaemia-induced endothelial ER stress by sulfhydrating protein disulphide isomerase to attenuate atherosclerosis

Shan Jiang et al. J Cell Mol Med. 2021 Apr.

Abstract

Hyperhomocysteinaemia (HHcy)-impaired endothelial dysfunction including endoplasmic reticulum (ER) stress plays a crucial role in atherogenesis. Hydrogen sulphide (H2 S), a metabolic production of Hcy and gasotransmitter, exhibits preventing cardiovascular damages induced by HHcy by reducing ER stress, but the underlying mechanism is unclear. Here, we made an atherosclerosis with HHcy mice model by ApoE knockout mice and feeding Pagien diet and drinking L-methionine water. H2 S donors NaHS and GYY4137 treatment lowered plaque area and ER stress in this model. Protein disulphide isomerase (PDI), a modulation protein folding key enzyme, was up-regulated in plaque and reduced by H2 S treatment. In cultured human aortic endothelial cells, Hcy dose and time dependently elevated PDI expression, but inhibited its activity, and which were rescued by H2 S. H2 S and its endogenous generation key enzyme-cystathionine γ lyase induced a new post-translational modification-sulfhydration of PDI. Sulfhydrated PDI enhanced its activity, and two cysteine-terminal CXXC domain of PDI was identified by site mutation. HHcy lowered PDI sulfhydration association ER stress, and H2 S rescued it but this effect was blocked by cysteine site mutation. Conclusively, we demonstrated that H2 S sulfhydrated PDI and enhanced its activity, reducing HHcy-induced endothelial ER stress to attenuate atherosclerosis development.

Keywords: atherosclerosis; endoplasmic reticulum stress; homocysteine; hydrogen sulphide; protein disulphide isomerase; sulfhydration.

PubMed Disclaimer

Conflict of interest statement

The authors confirm that there are no conflicts of interest.

Figures

FIGURE 1
FIGURE 1
H2S donor treatment lowered plaque area and ER stress in atherosclerosis with HHcy mice. A, Aorta plaque area was measured by en‐face Oil‐red O staining. B, Aortic root plaque size was detected by H&E staining (bar = 500 μm). C, Serum total cholesterol, LDL‐cholesterol (LDL‐c), HDL‐cholesterol (HDL‐c) changes after H2S donor treatment. D, ER stress marker protein‐GRP78 immunohistochemical staining was assayed in aortic root plaque (bar = 20 μm). Small images in the left corner are overall perspective pictures
FIGURE 2
FIGURE 2
H2S donor reduced PDI protein expression in atherosclerotic plaques. A, Immunofluorescence staining showed the PDI (green) expression in CD31 (red)‐positive cells of plaque. Bar = 100 μm. B, PDI expression in α‐SMA (red)‐positive cells of plaque. Bar = 100 μm
FIGURE 3
FIGURE 3
H2S donor lowered Hcy‐induced ER stress in human aortic endothelial cell (HAEC). A, ER stress markers including early markers PDI and GRP78, late markers spliced XBP1 (XBP1s) and cleaved‐caspase 12 (c‐caspase 12) were assayed by Western blot after different concentration of DL‐Hcy (from 25 to 400 μM) treatment for 24 h. B, The ER stress markers change while DL‐Hcy (200 μM) treated HAEC for different times (from 2 to 24 h). C, H2S donor‐GYY4137 reduced ER stress markers induced by Hcy (200 μM for 24 h)
FIGURE 4
FIGURE 4
H2S rescued HHcy‐induced PDI activity inhibition. A, PDI activity assessment with oxidative‐PDI/reduction‐PDI by native‐polyacrylamide gel electrophoresis, after different concentration of DL‐Hcy treatment for 24 h. B, PDI activity changes after treating HAEC with DL‐Hcy for different times. C, GYY4137 action on HHcy‐induced PDI activity inhibition
FIGURE 5
FIGURE 5
Endogenous CSE/H2S system in PDI sulfhydration and PDI activity. A, In vivo and In vitro PDI sulfhydration were identified by biotin switch assay. B, PDI sulfhydration associated with PDI activity. C, PDI sulfhydration changes while overexpressed cystathionine γ lyase (CSE) or knockdown CSE by adenovirus. D, Overexpression or knockdown CSE‐associated PDI activity changes. E, HHcy and H2S donor (GYY4137) treatment‐induced PDI sulfhydration changes in HAECs
FIGURE 6
FIGURE 6
PDI sulfhydration sites and their activity. A, Two conserved cysteine‐terminal CXXC in humans, rat and mouse. B, H2S induced PDI sulfhydration changes by transfection wild‐type human PDI (pPDI) plasmid, mutation C53/57 (M1) plasmid, mutation 397/400 plasmid (M2) and mutation four cysteine site (M3) plasmid into HEK‐293 cells. C, H2S‐promoted PDI activity changes while mutation sulfhydration sites of PDI. D, H2S reduced HHcy‐induced ER stress while mutation sulfhydration sites of PDI. E, The relative protein expression of GRP78, XBP1s and cleaved‐caspase‐12. **P < 0.01, *P < 0.05. F, Schematic diagram of the present findings
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. McCully KS. Vascular pathology of homocysteinemia: implications for the pathogenesis of arteriosclerosis. Am J Pathol. 1969;56:111‐128. - PMC - PubMed
    1. Lai WK, Kan MY. Homocysteine‐induced endothelial dysfunction. Ann Nutr Metab. 2015;67:1‐12. - PubMed
    1. Zhang C, Cai Y, Adachi MT, et al. Homocysteine induces programmed cell death in human vascular endothelial cells through activation of the unfolded protein response. J Biol Chem. 2001;276:35867‐35874. - PubMed
    1. Kabil O, Banerjee R. Enzymology of H2S biogenesis, decay and signaling. Antioxid Redox Signal. 2014;20:770‐782. - PMC - PubMed
    1. Kanagy NL, Szabo C, Papapetropoulos A. Vascular biology of hydrogen sulfide. Am J Physiol Cell Physiol. 2017;312:C537‐C549. - PMC - PubMed

Publication types

-