Inhibitory effect of angiotensin II on the brown fat differentiation of rat bone marrow mesenchymal stem cells

doi:10.12307/2025.083

Abstract

Abstract: BACKGROUND: Bone marrow mesenchymal stem cells are one of the sources of adipocytes and express all renin-angiotensin system components, but the effect of angiotensin II on bone marrow mesenchymal stem cell differentiation into brown adipose tissue is not clear.
OBJECTIVE: To observe the effect of angiotensin II on bone marrow mesenchymal stem cell differentiation into brown adipose tissue and investigate the role of angiotensin 1a receptor knockout in effect of angiotensin II on bone marrow mesenchymal stem cell differentiation into brown adipocytes and its potential mechanisms.
METHODS: After isolation and culture of bone marrow mesenchymal stem cells in wild-type and angiotensin 1a receptor knockout SD rats, the cells were cultured to the third generation and randomly divided into four groups: wild type group, knockout group, wild type + angiotensin II group, and knockout + angiotensin II group. The differentiation was induced in the brown fat induced differentiation medium for 14 days. Angiotensin II (100 nmol/L) was added for intervention when the differentiation medium was changed each time in the latter two groups. Western blot assay, qRT-PCR, immunofluorescence, and other methods were used to detect the expression of induced differentiation, lipolysis, β oxidation, and mitochondrial biogenesis in brown fat.

RESULTS AND CONCLUSION: Angiotensin II could inhibit the browning of rat bone marrow mesenchymal stem cells. Knockout of angiotensin 1a receptor could improve the inhibitory effect of angiotensin II on brown lipid formation of rat bone marrow mesenchymal stem cells by promoting lipolysis, enhancing fatty acid β oxidation, promoting mitochondrial biogenesis, and enhancing mitochondrial function. These findings provide new research directions and potential therapeutic targets for obesity treatment, revealing the important role of renin angiotensin systems in fat metabolism and its potential as a therapeutic target.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: angiotensin II, bone marrow mesenchymal stem cell, brown fat differentiation, mitochondria, β-oxidation, angiotensin II type 1a receptor

CLC Number:

Liu Chenyang, Wang Jin, Zhang Wenting, Wang Liqing, Yin Xiaoxiao, Zhao Junnan, Jiao Xiangying. Inhibitory effect of angiotensin II on the brown fat differentiation of rat bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(23): 4859-4867.

Figures/Tables 5

2.4 AT1aR敲除改善了AngⅡ对大鼠骨髓间充质干细胞棕色脂变过程中产热标记物表达的抑制作用产热在改善代谢率及葡萄糖和脂质代谢中发挥着重要作用[11]。除了上述棕色脂肪特异性标记蛋白UCP1和PPARγ外，PGC-1α是一种重要的转录共激活因子，被认为在能量代谢调控中发挥着关键作用[10]。PGC-1α也是棕色脂肪细胞中产热基因表达、线粒体生物发生和氧化代谢的主要转录调控因子[12]。FGF21作为一种转录因子和激素，是维持局部和全身脂质、葡萄糖和能量代谢稳态的主要调节因子，并且被认为是一种褐变剂[13]。DIO2是一种碘代谢酶，在棕色脂肪中促进甲状腺素的转化，从而增强脂肪细胞的产热能力[14]。CIDEA最初因与凋亡DNA分解因子序列的相似性而被识别出来，且其在棕色脂肪组织中表达水平很高[15]。如图4所示，与WT组相比，WT+Ang Ⅱ组大鼠骨髓间充质干细胞棕色脂肪变过程中产热标记物PGC-1α、CEBPα、DIO2、CIDEA的mRNA表达水平显著降低(P < 0.05)，AT1aR敲除组PGC-1α、CEBPα、DIO2、CIDEA、FGF21的mRNA表达水平均显著升高(P < 0.05)。与WT+Ang Ⅱ组相比，KO+Ang Ⅱ组的这些指标有明显升高(P < 0.05)。同时，PGC-1α的蛋白表达水平与qRT-PCR结果一致。以上结果表明，AT1aR敲除通过增加产热标记物的表达，进而改善Ang Ⅱ对大鼠骨髓间充质干细胞棕色脂变产热功能的不良影响。 2.5 AT1aR敲除改善了AngⅡ对大鼠骨髓间充质干细胞向棕色脂肪分化过程中脂解的抑制作用棕色脂肪组织增多，导致机体的产热增加，因而脂肪的分解和利用增多。如图5所示，与WT组相比，WT+AngⅡ组脂肪分解限速酶ATGL和HSL的mRNA表达水平显著降低(P < 0.05)，而AT1aR敲除组ATGL和HSL的mRNA表达水平显著升高(P < 0.05)。与WT+Ang Ⅱ组相比，KO+Ang Ⅱ组的这些指标有明显升高(P < 0.01)。与WT组相比，WT+Ang Ⅱ组ATGL的蛋白表达水平显著下降(P < 0.05)，而AT1aR敲除组ATGL的蛋白表达水平显著升高(P < 0.05)。与WT+Ang Ⅱ组相比，KO+Ang Ⅱ组ATGL的蛋白表达水平显著升高(P < 0.01)。 2.6 AT1aR敲除改善了Ang Ⅱ对大鼠骨髓间充质干细胞脂肪酸β氧化的抑制作用脂肪酸氧化相关酶包括ACADM、ACADL以及负责上游调节的PPAR家族[16]。有文献报道，促进游离脂肪酸代谢相关的脂肪酸β氧化速率是通过提高PPARα蛋白及其下游脂肪酸转位酶和肉碱棕榈酰基转移酶1蛋白的活性来发挥作用的[17]。在脂肪细胞中，CD36的主要作用是促进脂肪酸的摄取，它能够结合游离脂肪酸，使其进入细胞内，进而参与脂肪酸代谢[18]。如图6所示，与WT组相比，WT+Ang Ⅱ组PPARα、CPT1B、CD36、ACADM、ACADL的mRNA水平明显降低(P < 0.05)，AT1aR敲除组这些指标显著升高(P < 0.05)。同时，与WT+Ang Ⅱ组相比，KO+Ang Ⅱ组的这些指标明显升高(P < 0.01)。"

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