Exosomes derived from mesenchymal stem cell overexpressing periostin enhanced liver regeneration capacity

doi:10.3877/cma.j.issn.2095-1221.2025.02.001

Abstract

Abstract:

Objective

To investigate the promoting effect and mechanism of exosomes derived from mesenchymal stem cells （MSCs） overexpressing periostin （POSTN） on liver regeneration.

Methods

A 70 % partial hepatectomy （PHx） model was established in mice， and mice were administered PBS （PBS group）， MSCs-derived exosomes （MSC-Exo group）， or POSTN- overexpressing MSCs-derived exosomes （MSC-Exo^POSTN group） via tail vein injection after operation.Liver tissues were collected 72 h after operation.Hepatocyte proliferation ability was assessed by immunofluorescence staining for Ki-67 and proliferating cell nuclear antigen（PCNA）was detected by Western blot to evaluate hepatocyte proliferation.mRNA and protein levels of pro- regenerative factors （e.g.， HGF， IL-6） in liver tissues were measured via RT-qPCR and Western blot.A co-culture system of MSC-Exo^POSTN with AML-12 hepatocytes was established， and the expression of PCNA， PI3K， p-PI3K， p65， p-p65 were detected by Western blot and RT-qPCR.The difference between two groups was compared by independent samples t-test and the difference among multi-group was compared by one-way ANOVA.LSD-t test was used for pairwise comparison between different groups.

Results

Compared with PBS group， liver-to-body weight ratio ［（3.63 ±0.06）% vs （2.96 ± 0.08）%］， PCNA protein levels （0.62 ± 0.02 vs 0.32 ± 0.03）， Ki-67-positive cell ratio ［（39.33 ± 2.87）%vs （21.03 ± 1.63）%］， HGF mRNA （1.66 ± 0.06 vs 0.99 ± 0.09） and protein levels （1.98 ± 0.12 vs 1.00 ± 0.19）， and IL-6 mRNA （1.87 ± 0.07 vs 1.09 ± 0.12） and protein levels （1.51 ± 0.04 vs 0.97 ± 0.07） were increased in MSC-Exo^POSTN group （all P ＜ 0.05）.Compared with MSC-Exo group， liver-to-body weight ratio ［（3.63 ± 0.06）% vs （3.28 ± 0.06）%］， PCNA protein levels （0.62 ± 0.02 vs 0.51 ± 0.02）， Ki-67-positive cell ratio ［（39.33 ± 2.87）% vs （30.33 ±1.70）%］， HGF mRNA （1.66 ± 0.06 vs 1.32 ± 0.10） and protein levels （1.98 ± 0.12 vs 1.50 ± 0.09），and IL-6 mRNA （1.87 ± 0.07 vs 0.77 ± 0.08） and protein levels （1.51 ± 0.04 vs 0.79 ± 0.06） were increased in the MSC-Exo^POSTN group （all P ＜ 0.05）.In the MSC-Exo^POSTN and AML-12 hepatocyte co-culture system， compared with PBS group， the expression levels of PCNA（0.81 ± 0.06 vs 0.32 ±0.03） and p-p65 protein （1.15 ± 0.10 vs 0.81 ± 0.14） were increased in MSC-Exo^POSTN group （both P ＜ 0.05）， while p-PI3K level remained unchanged.Compared with MSC-Exo group， the expression levels of PCNA （0.81 ± 0.06 vs 0.49 ± 0.04） and p-p65 protein （1.15 ± 0.10 vs 0.51 ± 0.07） were increased in MSC-Exo^POSTN group （both P ＜ 0.05）， while p-PI3K level remained unchanged.Compared to PBS treatment， the PCNA mRNA level was reduced after treatment with NF-κB inhibitor JSH23 （1.01 ± 0.09 vs 1.59 ± 0.07， P ＜ 0.05）.

Conclusion

MSC-Exo^POSTN enhances liver regeneration post- hepatectomy， potentially via activation of the NF-κB signaling pathway in hepatocytes.

Key words: Partial hepatectomy, Mesenchymal stem cells, Exosomes, Liver regeneration, Hepatocyte growth factor

Jianhao Zhang, Danwen Cai, Chenhao Jiang, Yujun Zhang, Lu Han, Xuegang Zhao, Xin Lv, Jiaqi Xiao, Jiebin Zhang, Xin Sui, Yingcai Zhang. Exosomes derived from mesenchymal stem cell overexpressing periostin enhanced liver regeneration capacity[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2025, 15(02): 65-74.

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URL: https://zhxbygxbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-1221.2025.02.001

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Figures/Tables 12

References 26

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基因	序列（ 5' → 3'）	预期扩增产物长度（ bp）
POSTN	上游TGGTATCAAGGTGCTATCTGCG	135
	下游AATGCCCAGCGTGCCATAA
VEGF	上游TTTGCCAATCACACTTCCTGC	102
	下游ACACTGTGGTAATGTTGCTGG
HGF	上游ACTTCTGCCGGTCCTGTTG	222
	下游CCCCTGTTCCTGATACACCT
IL-6	上游CAACGATGATGCACTTGCAGA	201
	下游TCTCTCTGAAGGACTCTGGCT
PCNA	上游TTGCACGTATATGCCGAGACC	183
	下游GGTGAACAGGCTCATTCATCTCT
GAPDH	上游AATGGATTTGGACGCATTGGT	213
	下游TTTGCACTGGTACGTGTTGAT

基因	序列（ 5' → 3'）	预期扩增产物长度（ bp）
POSTN	上游TGGTATCAAGGTGCTATCTGCG	135
	下游AATGCCCAGCGTGCCATAA
VEGF	上游TTTGCCAATCACACTTCCTGC	102
	下游ACACTGTGGTAATGTTGCTGG
HGF	上游ACTTCTGCCGGTCCTGTTG	222
	下游CCCCTGTTCCTGATACACCT
IL-6	上游CAACGATGATGCACTTGCAGA	201
	下游TCTCTCTGAAGGACTCTGGCT
PCNA	上游TTGCACGTATATGCCGAGACC	183
	下游GGTGAACAGGCTCATTCATCTCT
GAPDH	上游AATGGATTTGGACGCATTGGT	213
	下游TTTGCACTGGTACGTGTTGAT

分组	实验次数	HGF mRNA	HGF 蛋白	IL-6 mRNA	IL-6 蛋白	VEGF mRNA	VEGF 蛋白
PBS	3	0.99 ± 0.09	1.00 ± 0.19	1.09 ± 0.12	0.97 ± 0.07	1.00 ± 0.20	1.00 ± 0.20
MSC-Exo	3	1.32 ± 0.10^a	1.50 ± 0.09^a	0.77 ± 0.08^a	0.79 ± 0.06^a	1.19 ± 0.05	1.03 ± 0.17
MSC-Exo^POSTN	3	1.66 ± 0.06^ab	1.98 ± 0.12^ab	1.87 ± 0.07^ab	1.51 ± 0.04^ab	1.21 ± 0.07	1.08 ± 0.14
F值		46.720	23.880	75.980	82.340	2.249	0.100
P值		＜ 0.001	＜ 0.001	＜ 0.001	＜ 0.001	0.187	0.907

分组	实验次数	HGF mRNA	HGF 蛋白	IL-6 mRNA	IL-6 蛋白	VEGF mRNA	VEGF 蛋白
PBS	3	0.99 ± 0.09	1.00 ± 0.19	1.09 ± 0.12	0.97 ± 0.07	1.00 ± 0.20	1.00 ± 0.20
MSC-Exo	3	1.32 ± 0.10^a	1.50 ± 0.09^a	0.77 ± 0.08^a	0.79 ± 0.06^a	1.19 ± 0.05	1.03 ± 0.17
MSC-Exo^POSTN	3	1.66 ± 0.06^ab	1.98 ± 0.12^ab	1.87 ± 0.07^ab	1.51 ± 0.04^ab	1.21 ± 0.07	1.08 ± 0.14
F值		46.720	23.880	75.980	82.340	2.249	0.100
P值		＜ 0.001	＜ 0.001	＜ 0.001	＜ 0.001	0.187	0.907

分组	实验次数	PCNA/α-tubulin	p-p65/p65	p-PI3K/PI3K
PBS	3	0.32 ± 0.03	0.81 ± 0.14	1.32 ± 0.06
MSC-Exo	3	0.49 ± 0.04^a	0.51 ± 0.07^a	1.41 ± 0.05
MSC-Exo^POSTN	3	0.81 ± 0.06^ab	1.15 ± 0.10^ab	1.42 ± 0.04
F值		57.970	26.360	2.693
P值		＜ 0.001	＜ 0.001	0.146

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