Tetramethylpyrazine improves the structure and function of mitochondrial-associated endoplasmic reticulum membrane and liver fibrosis

Introduction: Morphological and functional abnormalities of mitochondrial-associated endoplasmic reticulum (ER) membrane (MAM) have emerged as a key mediator of organelle dysfunction during liver fibrosis. Tetramethylpyrazine (TMP) was investigated as a potential therapy for liver fibrosis with an unclear mechanism.

Objectives: Considering the changes of MAM quantity and gap distance during liver fibrosis, we aimed to investigate the underlying mechanisms and their potential as therapeutic targets for TMP in inhibiting liver fibrosis.

Methods: Through different Sequencing techniques and a series of Molecular Biology experiments, we explored the effects and mechanisms of TMP in CCl₄-induced fibrosis models both in vivo and in vitro and examined key signaling in patients with fibrosis.

Results: An aberrant increase in the numbers of MAM and drastic alterations in the morphology of ER and mitochondria were accompanied by a substantial influx of CA²⁺ from the ER into mitochondria under fibrotic conditions. These changes were largely restored by TMP. Further isolation of distinct cellular fractions revealed that CCl₄ caused mis-localization and local concentration of MAM proteins, primarily by suppressing mitofusin 2 (MFN2). TMP directly bound to and stimulated MFN2 expression by activating transcription and inhibiting K79 ubiquitination-mediated degradation, which promoted the interaction and function of MFN2-sarco/endoplasmic reticulum CA²⁺ ATPase (SERCA2) complex for reversing CA²⁺ overload in mitochondria. Notably, findings in fibrosis patients and hepatic MFN2 knockdown mice further underscored the crucial role of MFN2-mediated normalization of MAM in improving liver fibrosis and the therapeutic effects of TMP.

Conclusion: Here, we highlight the therapeutic potential of TMP in liver fibrosis by elucidating its role in repairing hepatic MAM.

Liver fibrosis; MAM; MFN2; SERCA2.