肿节风总黄酮影响骨髓间充质干细胞及其外泌体促进巨核细胞分化的作用及机制

doi:10.3969/j.issn.1006-5725.2025.11.003

Abstract

Abstract:

Objective To investigate the effects and underlying mechanisms of total flavonoids of sarcandra glabra （TFFSG） on bone marrow mesenchymal stem cells （BMSCs） and their derived exosomes in immune thrombocytopenia （ITP）， with a focus on promoting megakaryocyte differentiation and maturation. Methods BMSCs induced by rabbit anti?rat platelet serum （APS） were divided into five groups： a blank control group， an ITP?BMSCs model group， and three TFFSG intervention groups with low （1.95 μg/mL）， medium （3.90 μg/mL）， and high doses （7.80 μg/mL）. The apoptosis rates and the expression levels of apoptosis?related proteins?B?cell lymphoma 2 （Bcl?2）， Bcl?2?associated X protein （BAX）， and Cysteinyl aspartate?specific proteinase?3 （Caspase?3）?were assessed. Exosomes were isolated from the blank control group （NC?BMSCs?Exos）， the ITP?BMSCs model group （ITP?BMSCs?Exos）， and the medium?dose TFFSG group （TFFSG?BMSCs?Exos）. Each group's exosomes （5 μg/mL） were co?cultured with megakaryocytic lineage Dami cells for 96 hours. Flow cytometry was employed to evaluate the expression of megakaryocytic differentiation markers （CD41a， CD42b， CD61） and the proportion of polyploid cells （≥ 4 N） in each group. Western Blot analysis was conducted to examine the expression of p?MEK1/2， MEK1/2， p?ERK1/2， and ERK1/2 across all groups. Results Compared with the ITP?BMSCs model group， the apoptosis rates in all TFFSG intervention groups were significantly reduced （P < 0.01）. In the medium? and high?dose TFFSG groups， BAX and Caspase?3 expression levels were markedly downregulated， whereas Bcl?2 expression was upregulated （P < 0.05， P < 0.01）. Compared with the ITP?BMSCs?Exos group， the TFFSG?BMSCs?Exos group demonstrated increased expression of CD41a⁺， CD42b⁺， and CD61⁺， a higher proportion of polyploid cells （≥ 4 N）（P < 0.05）， as well as elevated ratios of p?MEK1/2 to MEK1/2 and p?ERK1/2 to ERK1/2（P < 0.05）. Conclusion TFFSG inhibits apoptosis of ITP?state BMSCs in vitro and promotes megakaryocyte differentiation and polyploidization maturation through BMSC?derived exosomes by activating the MEK1/2?ERK1/2 signaling pathway.

Key words: immune thrombocytopenia, bone marrow mesenchymal stem cells, exosomes, total flavonoids of Sarcandra glabra

CLC Number:

285.5

Huizhen LIU,Xiaonan LU,Ge LIU,Guanqing CAI,Pingan LI,Yingjian ZENG,Guangbin SHANG. The effects and mechanism of total flavonoids of Sarcandra glabra in modulating bone marrow mesenchymal stem cells and their exosomes to promote megakaryocyte differentiation[J]. The Journal of Practical Medicine, 2025, 41(11): 1618-1626.

Figures/Tables 12

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References 27

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组别	24 h	48 h（%）	72 h	96 h
control组	100.00 ± 0.00	100.00 ± 0.00	100.00 ± 0.00	100.00 ± 0.00
0.975 μg/mL组	95.79 ± 4.31^?	98.87 ± 3.96	104.4 ± 4.28	95.97 ± 4.07
1.95 μg/mL组	97.29 ± 4.00	101.5 ± 4.20	108.9 ± 4.24^??	103.8 ± 3.77
3.9 μg/mL组	96.89 ± 2.88	98.24 ± 4.15	109.1 ± 4.79^??	102.8 ± 4.03
7.8 μg/mL组	94.05 ± 2.72^??	94.68 ± 3.28^??	108.0 ± 5.66^??	106.2 ± 5.98^?
15.6 μg/mL组	89.44 ± 2.73^???	83.75 ± 3.95^???	91.86 ± 2.89^??	76.23 ± 3.10^???
F值	7.988	19.79	16.54	39.13
P值	0.0001	< 0.0001	< 0.0001	< 0.0001

组别	剂量/（μg/mL）	早期凋亡率/%	晚期凋亡率/%	总凋亡率/%
正常对照组	-	5.71 ± 0.52	4.70 ± 0.16	10.55 ± 0.71
ITP-BMSCs模型组	-	31.88 ± 4.99^**	16.13 ± 1.60^**	45.23 ± 0.70^**
TFFSG低剂量组	1.95	13.43 ± 3.01^##	17.33 ± 1.97	30.77 ± 2.32^##
TFFSG中剂量组	3.90	14.93 ± 0.47^##	8.20 ± 0.92^##	23.14 ± 0.65^##
TFFSG高剂量组	7.80	8.61 ± 0.78^##	8.44 ± 0.18^##	17.05 ± 0.61^##
F值		43.74	61.39	375.0
P值		< 0.0001	< 0.0001	< 0.0001

组别	BAX	Bcl-2	Caspase-3
正常对照组	1 ± 0	1 ± 0	1 ± 0
ITP-BMSCs模型组	2.34 ± 0.53	0.36 ± 0.14^?	1.38 ± 0.04^?
TFFSG低剂量组	1.74 ± 0.35	0.52 ± 0.06	1.03 ± 0.08
TFFSG中剂量组	1.43 ± 0.22^#	0.70 ± 0.13	0.80 ± 0.20^#
TFFSG高剂量组	1.25 ± 0.31^#	1.06 ± 0.30^##	0.70 ± 0.23^#
F值	4.837	6.926	6.580
P值	0.057 1	0.028 5	0.031 6

组别	CD41a	CD42b	CD61
NC-BMSCs-Exos组	6.85 ± 0.40	20.47 ± 2.05	22.20 ± 1.71
APS-BMSCs-Exos组	4.00 ± 0.70	9.29 ± 0.07^?	2.51 ± 0.22^?
TFFSG-BMSCs-Exos组	11.47 ± 2.20^#?	26.93 ± 0.81^#?	27.07 ± 0.65^#?
TFFSG-BMSCs-Exos+抑制剂组	0.58 ± 0.14^&	7.62 ± 0.48^&	0.27 ± 0.21^&
F值	46.44	199.6	645.6
P值	< 0.000 1	< 0.000 1	< 0.000 1

组别	2N	≥4N
NC-BMSCs-Exos组	54.57 ± 1.00	39.63 ± 0.51
APS-BMSCs-Exos组	63.10 ± 4.02^?	28.13 ± 1.10^?
TFFSG-BMSCs-Exos组	46.13 ± 0.12^#?	50.40 ± 1.14^#?
TFFSG-BMSCs-Exos+抑制剂组	57.33 ± 2.21^&	20.87 ± 1.69^&
F值	27.13	360.3
P值	0.000 2	< 0.000 1

The effects and mechanism of total flavonoids of Sarcandra glabra in modulating bone marrow mesenchymal stem cells and their exosomes to promote megakaryocyte differentiation

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组别	p-MEK1/2/MEK1/2	p-ERK1/2/ERK1/2
NC-BMSCs-Exos组	1 ± 0	1 ± 0
APS-BMSCs-Exos组	0.63 ± 0.18^?	0.57 ± 0.03^?
TFFSG-BMSCs-Exos组	1.47 ± 0.12^#	1.05 ± 0.19^#*
TFFSG-BMSCs-Exos+抑制剂组	0.16 ± 0.15^&	0.17 ± 0.01^&
F值	53.71	54.59
P值	< 0.000 1	< 0.000 1