Effects of human umbilical cord-derived mesenchymal stem cells on chronic intermittent hypoxia in mice

doi:10.11958/20250965

Abstract

Abstract:

Objective To investigate the therapeutic potential of human umbilical cord-derived mesenchymal stem cells (hUCMSCs) in modulating the cGAS-STING-NF-κB signaling pathway in chronic intermittent hypoxia (CIH) mice. Methods Twenty-four C57BL/6 mice were divided into the control group, the model group, the hUCMSCs group and the hUCMSCs+STING agonist (DMXAA) group, with 6 mice in each group. Except for the control group, the other groups were exposed to hypoxic conditions for 8 hours daily for a total of 8 weeks to establish the CIH mouse model. After 8 weeks, mice were anesthetized for cardiac blood collection followed by euthanasia and lung tissue collection. Serum levels of IL-6, TNF-α, IL-1β and IL-17A were measured by ELISA. Pulmonary inflammatory infiltration and collagen deposition were assessed by HE and Masson staining. E-Cadherin and α-SMA expression levels were evaluated by immunohistochemistry. Expression levels of cGAS, STING and NF-κB mRNA were detected by RT-qPCR, while protein expression levels of E-Cadherin, N-Cadherin, α-SMA, Vimentin, cGAS, STING and NF-κB were analyzed by Western blot assay. Results Compared with the control group, levels of IL-6, IL-1β, TNF-α and IL-17A increased in the model group, inflammation and fibrosis scores increased, mRNA expression levels of cGAS, STING and NF-κB increased, and protein expression levels of N-Cadherin, α-SMA, Vimentin, cGAS, STING and NF-κB increased. In contrast, E-Cadherin protein expression was significantly decreased (P<0.05). Compared with the model group, IL-6, IL-1β, TNF-α and IL-17A decreased in the hUCMSCs group, mRNA expression levels of cGAS, STING and NF-κB were decreased, protein expression levels of N-Cadherin, α-SMA, Vimentin, cGAS, STING and NF-κB were also decreased. Meanwhile, E-Cadherin protein expression was significantly increased (P<0.05). STING activator DMXAA reversed the protective effects of hUCMSCs in CIH mice (P<0.05). Conclusion Intravenous administration of hUCMSCs alleviates pulmonary inflammatory infiltration and epithelial-mesenchymal transition in mouse model of intermittent hypoxia, which may be related to the down-regulation of the cGAS-STING-NF-κBsignaling pathway.

Key words: mesenchymal stem cells, hypoxia, inflammation, epithelial-mesenchymal transition, signal transduction, chronic intermittent hypoxia

CLC Number:

R329.2

Figures/Tables 9

References 21

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基因名称	引物序列（5′→3′）		产物大小/bp
cGAS	上游：	GAGGCGCGGAAAGTCGTAA	98
	下游：	TTGTCCGGTTCCTTCCTGGA	98
STING	上游：	GGTCACCGCTCCAAATATGTAG	135
	下游：	CAGTAGTCCAAGTTCGTGCGA	135
NF-κB	上游：	AGGCTTCTGGGCCTTATGTG	111
	下游：	TGCTTCTCTCGCCAGGAATAC	111
GAPDH	上游：	AGGTCGGTGTGAACGGATTTG	123
	下游：	TGTAGACCATGTAGTTGAGGTCA	123

基因名称	引物序列（5′→3′）		产物大小/bp
cGAS	上游：	GAGGCGCGGAAAGTCGTAA	98
	下游：	TTGTCCGGTTCCTTCCTGGA	98
STING	上游：	GGTCACCGCTCCAAATATGTAG	135
	下游：	CAGTAGTCCAAGTTCGTGCGA	135
NF-κB	上游：	AGGCTTCTGGGCCTTATGTG	111
	下游：	TGCTTCTCTCGCCAGGAATAC	111
GAPDH	上游：	AGGTCGGTGTGAACGGATTTG	123
	下游：	TGTAGACCATGTAGTTGAGGTCA	123

组别	IL-6	IL-1β
对照组	28.702±6.745	6.974±2.048
模型组	119.202±31.029^a	39.970±5.659^a
hUCMSCs组	41.570±11.875^b	20.756±6.226^b
hUCMSCs+DMXAA组	167.679±25.910^c	33.981±4.150^c
F	57.070^＊	56.140^＊
组别	TNF-α	IL-17A
对照组	6.542±1.511	7.042±0.793
模型组	23.165±4.200^a	23.900±8.091^a
hUCMSCs组	12.028±1.495^b	10.428±2.030^b
hUCMSCs+DMXAA组	24.743±3.744^c	27.427±2.267^c
F	51.183^＊	31.695^＊

组别	IL-6	IL-1β
对照组	28.702±6.745	6.974±2.048
模型组	119.202±31.029^a	39.970±5.659^a
hUCMSCs组	41.570±11.875^b	20.756±6.226^b
hUCMSCs+DMXAA组	167.679±25.910^c	33.981±4.150^c
F	57.070^＊	56.140^＊
组别	TNF-α	IL-17A
对照组	6.542±1.511	7.042±0.793
模型组	23.165±4.200^a	23.900±8.091^a
hUCMSCs组	12.028±1.495^b	10.428±2.030^b
hUCMSCs+DMXAA组	24.743±3.744^c	27.427±2.267^c
F	51.183^＊	31.695^＊

组别	炎症评分	纤维化评分
对照组	0.958±0.246	0.833±0.204
模型组	3.188±0.360^a	1.729±0.399^a
hUCMSCs组	1.117±0.229^b	1.167±0.303^b
hUCMSCs+DMXAA组	2.729±0.544^c	1.708±0.332^c
F	44.153^＊	11.373^＊