Indole-3-carboxamide mitigating myocyte atrophy by the transforming growth factor-β1/activin receptor ⅡB pathway

施恒 吴成林 刘亚茹 刘知非 周兆丽   陆秀宏

doi:10.16098/j.issn.0529-1356.2026.02.007

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Acta Anatomica Sinica ›› 2026, Vol. 57 ›› Issue (2) : 194-201. DOI: 10.16098/j.issn.0529-1356.2026.02.007

Cell and Molecules Biology

Indole-3-carboxamide mitigating myocyte atrophy by the transforming growth factor-β1/activin receptor ⅡB pathway

SHI Heng¹, WU Cheng-lin², LIU Ya-ru¹, LIU Zhi-fei², ZHOU Zhao-li^1,3*, LU Xiu-hong^1,2*

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Abstract

Objective To evaluate the alleviating effect of indole-3-carboxamide(WCL-3) on myocyte atrophy in vitro. Methods An activin receptor ⅡB (ActRⅡB) kinase inhibition assay in vitro was established to assess the inhibitory activity of WCL-3. Based on the enzymatic inhibition result, WCL-3 anti-muscle atrophy and anti-adipocyte degradation effects were further investigated. Myotube diameter measurement and morphological observation were used to evaluate the protective effect of WCL-3 on myotube atrophy. Oil red O staining and quantitative analysis were conducted to assess the WCL-3 inhibitory effect on adipocyte degradation. Western blotting analysis was performed to detect the expression levels of p-Smad2/3, Smad2/3, p-Akt, Akt, Atrogin-1, and myogenic differentiation 1(MyoD1) in muscle cells. Results Kinase inhibition assays demonstrated that WCL-3 exhibited potential inhibition of ActRⅡB kinase, half-maxinal inhibitory concentration (IC50)=4.1 nmol/L. In C2C12 myotube atrophy models induced by CT26 tumor-conditioned medium and transforming growth factor-β1（TGF-β1）, WCL-3 alleviated myotube atrophy in a dose-dependent manner. At the molecular level, WCL-3 downregulated the phosphorylation of Smad2/3 and inhibited the expression of Atrogin-1, while upregulating p-Akt and promoting MyoD1 synthesis, thereby attenuating muscle atrophy. Additionally, WCL-3 ameliorated CT26-conditioned medium-induced lipid degradation in mature 3T3-L1 adipocytes. Conclusion WCL-3 significantly alleviates myocyte atrophy in vitro primarily by inhibiting the TGF-β/ActRⅡB signaling pathway. Therefore, WCL-3 represents a promising lead compound for the development of therapeutic agents targeting muscle wasting and fat loss.

Key words

Myocyte atrophy

/ Adipocyte degradation / Transforming growth factor-β1 / Activin receptor type ⅡB / Indole-3-carboxamide / Western blotting

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SHI Heng, WU Cheng-lin, LIU Ya-ru, LIU Zhi-fei, ZHOU Zhao-li, LU Xiu-hong. Indole-3-carboxamide mitigating myocyte atrophy by the transforming growth factor-β1/activin receptor ⅡB pathway[J]. Acta Anatomica Sinica. 2026, 57(2): 194-201 https://doi.org/10.16098/j.issn.0529-1356.2026.02.007

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