Effect of behavioral characteristics in smooth muscle cells deriving from the human great saphenous veins underlying hypoxia

doi:10.3877/cma.j.issn.1674-0793.2025.03.004

Abstract

Abstract:

Objective

To establish a simulated hypoxia model of venous smooth muscle cells（SMCs）， and to investigate the effect of hypoxia on the behaviorial characteristics of SMCs derived from the human normal great saphenous veins （GSVs）.

Methods

Fifteen tissue samples were collected from patients undergoing distal arterial bypass grafting surgery without GSVs injuries in the 80th Group Army Hospital of the People’s Liberation Army from May 2020 to May 2021.SMCs were isolated and cultured， then the cobalt chloride （CoCl₂） group and the control group were assigned.Using specific staining， proliferation，migration， adhesion， senescence and the structure of cytoskeletal filaments in SMCs were observed.mRNA and protein expression of Bax， Bcl-2， caspase-3， matrix metalloproteinases （MMP）-2， MMP-9， tissue inhibitor of metalloproteinases （TIMP）-1， and TIMP-2 were detected by fluorescent quantitative polymerase chain reaction and immunoblotting.

Results

The quantity of adhesion， migration， senescence， proliferation and mRNA and protein expression of Bcl-2， MMP-2， MMP-9， TIMP-1， and TIMP-2 in SMCs in the CoCl₂ group were significantly increased compared with the control group （t=5.994， 5.687， 13.898， 25.979，11.704， 20.705， 16.363， 12.228， 11.473， 9.618， 14.780， 14.691， 64.185， 14.915， all P＜0.05）.mRNA and protein expression of Bax and caspase-3 were significantly decreased （t=27.297， 15.430， 17.851， 17.801， all P＜0.05）.

Conclusion

Under hypoxic conditions， the phenotype and function of SMCs derived from the human NGSVs are dysregulated， suggesting that SMCs switch from the contractile phenotype to the secretory or synthetic phenotype， and more dedifferentiate， and apoptotic decrease through the intrinsic pathway.

Key words: Varicose great saphenous veins, Smooth muscle cells, Behavior, Hypoxia

Jianhua Ju, Shunchang Zhou, Yongbo Xu, Xiaoxuan Hu, Yuxu Zhong, Haibo Chu. Effect of behavioral characteristics in smooth muscle cells deriving from the human great saphenous veins underlying hypoxia[J]. Chinese Archives of General Surgery(Electronic Edition), 2025, 19(03): 169-174.

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Figures/Tables 11

References 25

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基因	引物序列	产物大小（bp）
Bax	上游引物：5-ATCTTTGCTGGAGACAAATTCTGGA-3	139
	下游引物：5-GCTTCAGGTAATAGGCACCCTTGA-3
Bcl-2	上游引物：5-CCTGTGGATGACTGAGTACCTGAAC-3	139
	下游引物：5-CAGAGTCTTCAGAGACAGCCAGGA-3
caspase-3	上游引物：5-GACTCTGGAATATCCCTGGACAACA-3	140
	下游引物：5-AGGTTTGCTGCATCGACATCTG-3
MMP-2	上游引物：5-ATCTTTGCTGGAGACAAATTCTGGA-3	175
	下游引物：5-GCTTCAGGTAATAGGCACCCTTGA-3
MMP-9	上游引物：5-ACGCACGACGTCTTCCAGTA-3	94
	下游引物：5-CCACCTGGTTCAACTCACTCC-3
TIMP-1	上游引物：5-CCTTATACCAGCGTTATGAGATCAA-3	194
	下游引物：5-AGTGATGTGCAAGAGTCCATCC-3
TIMP-2	上游引物：5-CTGGACGTTGGAGGAAAGAAGG-3	148
	下游引物：5-CCATCTGGTACCTGTGGTTCAGG-3
GAPDH	上游引物：5-GCACCGTCAAGGCTGAGAAC-3	138
	下游引物：5-TGGTGAAGACGCCAGTGGA-3

基因	引物序列	产物大小（bp）
Bax	上游引物：5-ATCTTTGCTGGAGACAAATTCTGGA-3	139
	下游引物：5-GCTTCAGGTAATAGGCACCCTTGA-3
Bcl-2	上游引物：5-CCTGTGGATGACTGAGTACCTGAAC-3	139
	下游引物：5-CAGAGTCTTCAGAGACAGCCAGGA-3
caspase-3	上游引物：5-GACTCTGGAATATCCCTGGACAACA-3	140
	下游引物：5-AGGTTTGCTGCATCGACATCTG-3
MMP-2	上游引物：5-ATCTTTGCTGGAGACAAATTCTGGA-3	175
	下游引物：5-GCTTCAGGTAATAGGCACCCTTGA-3
MMP-9	上游引物：5-ACGCACGACGTCTTCCAGTA-3	94
	下游引物：5-CCACCTGGTTCAACTCACTCC-3
TIMP-1	上游引物：5-CCTTATACCAGCGTTATGAGATCAA-3	194
	下游引物：5-AGTGATGTGCAAGAGTCCATCC-3
TIMP-2	上游引物：5-CTGGACGTTGGAGGAAAGAAGG-3	148
	下游引物：5-CCATCTGGTACCTGTGGTTCAGG-3
GAPDH	上游引物：5-GCACCGTCAAGGCTGAGAAC-3	138
	下游引物：5-TGGTGAAGACGCCAGTGGA-3

组别	Bax		Bcl-2		caspase-3
组别	基因	蛋白	基因	蛋白	基因	蛋白
CoCl₂组	0.69±0.045^a	0.95±0.181^a	1.43±0.144^a	1.35±0.119^a	0.81±0.047^a	0.35±0.048^a
对照组	1.00±0.000	2.03±0.147	1.00±0.000	0.59±0.115	1.00±0.000	0.64±0.105
t值	27.297	17.851	11.704	9.618	15.430	17.801
P值	＜0.001	＜0.001	＜0.001	＜0.001	＜0.001	＜0.001

组别	Bax		Bcl-2		caspase-3
组别	基因	蛋白	基因	蛋白	基因	蛋白
CoCl₂组	0.69±0.045^a	0.95±0.181^a	1.43±0.144^a	1.35±0.119^a	0.81±0.047^a	0.35±0.048^a
对照组	1.00±0.000	2.03±0.147	1.00±0.000	0.59±0.115	1.00±0.000	0.64±0.105
t值	27.297	17.851	11.704	9.618	15.430	17.801
P值	＜0.001	＜0.001	＜0.001	＜0.001	＜0.001	＜0.001

组别	MMP-2		MMP-9		TIMP-1		TIMP-2
组别	基因	蛋白	基因	蛋白	基因	蛋白	基因	蛋白
CoCl₂ 组	1.79±0.148^a	0.90±0.082^a	1.64±0.151^a	0.65±0.090^a	1.51±0.162^a	1.45±0.064^a	1.38±0.129^a	0.80±0.062^a
对照组	1.00±0.000	0.44±0.088	1.00±0.000	0.25±0.055	1.00±0.000	0.17±0.042	1.00±0.000	0.48±0.057
t值	20.705	14.780	16.363	14.691	12.228	64.185	11.473	14.915
P值	＜0.001	＜0.001	＜0.001	＜0.001	＜0.001	＜0.001	＜0.001	＜0.001

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