Expression of functional thyroid stimulating hormone receptor in human microvascular endothelial cells

doi:10.3877/cma.j.issn.2095-655X.2018.01.015

Abstract

Abstract:

Objective

To investigate the expression and activity of thyroid stimulating hormone receptor (TSHR) in human microvascular endothelial cells(HMEC-1).

Methods

Culture HMEC-1 cells in vitro and detected TSHR mRNA by reverse transcription-polymerase chain reaction(RT-PCR). The cells were seeded on the six-well plate and divided into blank control group, low dose group, middle dose group and high dose group.The blank control group was cultured with basal medium without thyroid stimulating hormone(TSH). The low dose group, middle dose group and high dose group were given 0.10 U/L, 1.00 U/L, 10.00 U/L TSH intervention, respectively, and after 24 h, the intracellular cyclic adenosine monophosphate(cAMP) content was measured by enzyme-linked immunosorbent assay (ELISA) to study whether TSHR on HMEC-1 was active.Significant test was analysed by one-way anova, and LSD-t was used on amparion among groups.

Results

RT-PCR results showed that HMEC-1 expressed TSHR mRNA.ELISA results showed that the expression of cAMP increased in HMEC-1 stimulated by TSH, there was significant difference between low dose group and blank control group [(5.23±0.86)mmol/L, (1.77±0.25)mmol/L, t=3.47, P<0.01], and this effect was dose-dependent (F=92.36, P<0.01).

Conclusion

TSHR is present in HMEC-1, and the receptor may interact specifically with TSH.

Key words: Receptors, thyrotropin, Microvessels, Endothelial cells

Qiang Meng, Minmin Shi, Jing Chen, Pengfei Yi, Na Wang, Lin Sun. Expression of functional thyroid stimulating hormone receptor in human microvascular endothelial cells[J]. Chinese Journal of Diagnostics(Electronic Edition), 2018, 06(01): 69-72.

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References 15

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引物名称	引物序列(5′－3′)	扩增产物大小(bp)
TSHR F	TGCCTTCGTCATCGTCTG	113
TSHR R	TCAACACAGCCATCCTCTTG	?
β-actin F	CCTGGCACCCAGCACAAT	144
β-actin R	GGGCCGGACTCGTCATAC	?

引物名称	引物序列(5′－3′)	扩增产物大小(bp)
TSHR F	TGCCTTCGTCATCGTCTG	113
TSHR R	TCAACACAGCCATCCTCTTG	?
β-actin F	CCTGGCACCCAGCACAAT	144
β-actin R	GGGCCGGACTCGTCATAC	?

组别	cAMP
空白对照组(0 U/L)	1.77±0.25
低剂量组(0.10 U/L)	5.23±0.86
中剂量组(1.00 U/L)	10.87±1.20
高剂量组(10.00 U/L)	16.10±1.71
F值	92.36
P值	<0.01

组别	cAMP
空白对照组(0 U/L)	1.77±0.25
低剂量组(0.10 U/L)	5.23±0.86
中剂量组(1.00 U/L)	10.87±1.20
高剂量组(10.00 U/L)	16.10±1.71
F值	92.36
P值	<0.01

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