Effects of chrysotile on expression of Wnt5a, p16 and p21 in endothelial cells

doi:10.11958/20240060

Abstract

Abstract:

Objective To investigate the influence of chrysotile on human umbilical vein endothelial cells (HUVECs). Methods HUVECs in the experimental group were treated with 50, 100 and 200 mg/L chrysotile for 24, 48 and 72 h. The control group was cultured with RPMI 1640 medium only. Morphological changes of HUVECs were observed. Cell viability was detected by MTT assay. β-Galactosidase staining was used to analyze the cell senescence. The cell survival rate was determined by tetramethylazolium blue assay. Wnt5a, p16 and p21 expression were investigated by qRT-PCR. Results Cells of the experimental group were oval and irregular, and some of them were round or irregular, with naked nuclei and vacuolar, and dead cells were present. With the increase of chrysotile concentration and exposure time, the level of cell viability decreased and the number of aged cells increased gradually (P<0.05). The cell growth of HUVECs was more active when 100 mg/L chrysotile treated for 24 h. Compared with the control group, the mRNA expression levels of Wnt5a, p16 and p21 were increased in the experimental group (P<0.05). Conclusion Chrysotile can promote HUVECs senescence, and Wnt5a, p16 and p21 are involved in this mechanism.

Key words: asbestos, serpentine, human umbilical vein endothelial cells, pulmonary fibrosis, Wnt-5a protein, genes, p16, genes, p21

CLC Number:

R563.9

Figures/Tables 6

References 20

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基因名称	引物序列（5’→3’）	产物大小/bp
Wnt5a	上游：CTAACTTAGCTGTGTGGGACATG	235
	下游：AAATGCAGAAAGCAAGCTAGCAG	235
p16	上游：CGGGGACATCAAGACATCGT	203
	下游：GCCGGATTTAGCTCTGCTCT	203
p21	上游：ACATCTCAGGGCCGAAAACG	173
	下游：AAGACACACAGAGTGAGGGC	173
β-actin	上游：CCACTGGCATCGTGATGGAC	171
	下游：GCGGATGTCCACGTCACACT	171

基因名称	引物序列（5’→3’）	产物大小/bp
Wnt5a	上游：CTAACTTAGCTGTGTGGGACATG	235
	下游：AAATGCAGAAAGCAAGCTAGCAG	235
p16	上游：CGGGGACATCAAGACATCGT	203
	下游：GCCGGATTTAGCTCTGCTCT	203
p21	上游：ACATCTCAGGGCCGAAAACG	173
	下游：AAGACACACAGAGTGAGGGC	173
β-actin	上游：CCACTGGCATCGTGATGGAC	171
	下游：GCGGATGTCCACGTCACACT	171

组别	24 h	48 h	72 h
对照组	4.20±0.84	9.20±1.48	11.80±1.92
50 mg/L实验组	8.20±1.92	13.20±2.39	17.40±3.05
100 mg/L实验组	11.40±1.52	17.20±2.86	28.60±3.91
200 mg/L实验组	22.00±4.36	29.60±3.58	37.60±6.42

组别	24 h	48 h	72 h
对照组	4.20±0.84	9.20±1.48	11.80±1.92
50 mg/L实验组	8.20±1.92	13.20±2.39	17.40±3.05
100 mg/L实验组	11.40±1.52	17.20±2.86	28.60±3.91
200 mg/L实验组	22.00±4.36	29.60±3.58	37.60±6.42

组别	Wnt5a	p16	p21
对照组	5.44±3.74	6.56±1.74	2.23±0.91
实验组	34.18±6.08	20.63±0.86	7.80±0.73
t	5.167^＊	9.881^＊	6.091^＊