人脐带间充质干细胞培养上清对米非司酮处理的人子宫内膜基质细胞存活、凋亡和子宫内膜容受性的影响

doi:10.13481/j.1671-587X.20240110

Abstract

Abstract:

Objective To discuss the effect of human umbilical cord mesenchymal stem cells culture supernatant （hUCMSCs-Sup） on the proliferation， apoptosis， and endometrium receptivity of the human endometrial stromal cells （hEndoSCs） treated with mifepristone （Ms）， and to clarify the possible mechanism. Methods The hEndoSCs were cultured in vitro and divided into control group and 40， 60， 80， and 100 μmol·L^－1 Ms groups. The survival rates of the cells in various groups were detected by MTT assay. The hEndoSCs were divided into control group， 40 μmol·L^－1 Ms group， and 60 μmol·L^－1 Ms group.The apoptotic rates of the cells in various groups were detected by flow cytometry； the expression levels of apoptosis-related protein B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax） proteins in the cells in various groups were detected by Western blotting method， and the ratio of Bcl-2/Bax was calculated. After treated with hUCMSCs-Sup， the hEndoSCs were divided into control group， Ms group， Ms+hUCMSCs-Sup group， and Ms+hUCMSCs-Sup+3-methyladenine （3-MA） group.The survival rates of the cells in various groups were detected by MTT assay；the apoptotic rates of the cells in various groups were detected by flow cytometry；the expression levels of microtubule-associated protein 1 light chain 3B-Ⅱ （LC3B-Ⅱ） and microtubule-associated protein 1 light chain 3B-I （LC3B-Ⅰ） proteins in the cells in various groups were detected by Western blotting method， and the ratio of LC3B-Ⅱ/LC3B-Ⅰ was calculated； the expression levels of endometrium receptivity marker molecules mRNA in the cells in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） method. Results Compared with control group， the survival rates of the cells in 40， 60， 80， and 100 μmol·L^－1 Ms groups were significantly decreased （P<0.05）in a time-dependent and dose-dependent manner. Compared with control group， the apoptotic rates of the cells in 40 and 60 μmol·L^－1 Ms groups were significantly increased （P<0.05）， and the ratios of Bcl-2/Bax were significantly decreased （P<0.05）. After treated with hUCMSCs-Sup， compared with control group， the survival rate of the cells and ratio of LC3B-Ⅱ/LC3B-Ⅰ in the cells in Ms group were significantly decreased （P<0.05）， the apoptotic rate was significantly increased （P<0.05）， and the expression levels of homeobox A10 （HOXA10）， leukemia inhibitory factor （LIF）， and integrin subunit beta 3 （ITGB3） mRNA in the cells were significantly decreased （P<0.05）；compared with Ms group， the survival rate of the cells and ratio of LC3B-Ⅱ/LC3B-Ⅰ in the cells in Ms+hUCMSCs-Sup group were significantly increased （P<0.05），the apoptotic rate was significantly decreased （P<0.05）， and the expression levels of HOXA10， LIF， and ITGB3 mRNA in the cells were significantly increased （P<0.05）； compared with Ms+hUCMSCs-Sup group， the survival rate of the cells and ratio of LC3B-Ⅱ/LC3B-Ⅰ in the cells in Ms+hUCMSCs-Sup+3-MA group were significantly decreased （P<0.05）. Conclusion hUCMSCs-Sup can increase the survival rate and decrease the apoptotic rate of the hEndoSCs after treated with Ms，and increase the endometrium receptivity，and its mechanism may be associated with the activation of autophagy of the hEndoSCs by hUCMSCs-Sup.

Key words: Umbilical cord mesenchymal stem cell, Human endometrial stromal cell, Apoptosis, Autophagy, Endometrium receptivity

CLC Number:

R711

Mengxue WU,Shiling CHEN,Yan LIU,Xuguang MI,Xiuying LIN,Jianhua FU,Yanqiu FANG. Effect of culture supernatant of human umbilical cord mesenchymal stem cells on survival，apoptosis and endometrium receptivity of human endometrial stromal cells after treated with mifepristone[J].Journal of Jilin University(Medicine Edition), 2024, 50(1): 79-87.

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References 31

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Group	Survival rate of cells
Group	(t/h) 0	24	48
Control	100.00±1.36	100.00±1.59	100.00±5.23
Ms(μmol·L^－1)
40	100.00±3.36	94.20±2.18^*△	69.30±1.42^*△#
60	100.00±4.27	69.73±2.09^*△	33.90±1.88^*△#
80	100.00±3.58	55.97±1.13^*△	22.11±2.43^*△#
100	100.00±6.20	54.22±1.02^*△	16.70±1.22^*△#

Group	Apoptotic rate（η/%）	Ratio of Bcl-2/Bax
Control	7.44±0.38	3.02±5.02
Ms(μmol·L^－1)
40	14.61±1.11^*	2.27±0.04^*
60	20.40±1.82^*	1.21±0.14^*

Group	Survival rate	Apoptotic rate
Control	100.00±3.41	6.60±1.25
Ms	68.83±1.47^*	20.43±0.71^*
Ms+hUCMSCs-Sup	85.00±1.41^*△	11.30±0.61^*△

Group	Survival rate (η/%)	Ratio of LC3B-Ⅱ/LC3B-Ⅰ
Control	100.00±1.29	0.39±0.10
Ms	67.12±1.85^*	0.19±0.02^*
Ms+hUCMSCs-Sup	84.67±2.32^△	1.32±0.57^△
Ms+hUCMSCs-Sup+3-MA	35.12±1.67^#	1.03±0.25^#

Group	HOXA10	LIF	ITGB3
Control	1.00±0.33	1.00±0.27	1.00±0.72
Ms	0.58±0.12^*	0.44±0.70^*	0.63±0.02^*
Ms+hUCMSCs-Sup	2.40±0.14^*△	1.93±0.07^*△	2.16±0.16^*△

Effect of culture supernatant of human umbilical cord mesenchymal stem cells on survival，apoptosis and endometrium receptivity of human endometrial stromal cells after treated with mifepristone

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