通过自体间充质干细胞实现肝硬化肝脏再生: 分子和病理生理机制
抽象的
Liver cirrhosis represents the terminal stage of chronic liver injury, 以不可逆纤维化为特征, 肝细胞损失, and vascular remodeling that culminate in hepatic failure. Conventional therapies can slow disease progression but rarely reverse parenchymal destruction. Recent advances in regenerative medicine have explored mesenchymal stem cells (间充质干细胞) as a promising therapeutic option to restore hepatic structure and function. This review summarizes the molecular and pathophysiological mechanisms underlying liver regeneration induced by autologous MSCs, focusing on their signaling interactions, 分化潜力, and paracrine effects. It also outlines the general laboratory processes for MSC isolation and ex vivo expansion commonly reported in clinical research settings. Collectively, these findings support the potential of MSC-based therapy as a disease-modifying approach for cirrhosis.
1. 介绍
Liver cirrhosis remains one of the leading causes of global mortality, affecting more than 120 million people worldwide. The hallmark of cirrhosis is the replacement of normal hepatic lobular architecture with fibrous septa and regenerative nodules. Although the liver possesses remarkable regenerative capacity, chronic insults—such as viral hepatitis, 酗酒, and nonalcoholic steatohepatitis—eventually exhaust endogenous repair mechanisms. 间充质干细胞 (间充质干细胞) have emerged as a viable approach to promote regeneration through paracrine signaling, anti-inflammatory effects, and differentiation into hepatocyte-like cells (HLCs).
MSCs can be obtained from various autologous sources—bone marrow, 脂肪组织, or peripheral blood—circumventing ethical and immunologic barriers associated with allogeneic or embryonic cells. The scientific basis for their therapeutic use lies in their immunomodulatory capacity and ability to home to injured tissues through chemokine signaling pathways such as SDF-1/CXCR4.
2. Pathophysiology of Liver Cirrhosis
Cirrhosis represents the end point of progressive hepatic fibrosis. Persistent hepatocyte injury activates Kupffer cells and hepatic stellate cells (造血干细胞), which transform into myofibroblast-like cells secreting extracellular matrix (细胞外基质) 蛋白质, primarily type I and III collagen. This ECM accumulation disrupts hepatic sinusoidal architecture and blood flow.
Inflammatory cytokines including tumor necrosis factor-α (肿瘤坏死因子-α), interleukin-1β, and transforming growth factor-β1 (TGF-β1) perpetuate stellate cell activation and suppress hepatocyte proliferation. Sinusoidal capillarization, 氧化应激, and mitochondrial dysfunction further contribute to hepatocellular loss. 最终, the liver loses its capacity for self-regeneration, resulting in portal hypertension and hepatic insufficiency.
3. Mechanisms of MSC-Mediated Liver Regeneration
3.1 Differentiation into Hepatocyte-Like Cells
MSCs exhibit multipotent differentiation potential. Under specific conditions—such as exposure to hepatogenic growth factors (肝细胞生长因子 [肝细胞生长因子], oncostatin M, fibroblast growth factor-4, and dexamethasone)—they can acquire morphological and functional characteristics of hepatocytes, including albumin secretion and cytochrome P450 enzyme activity. Studies have shown that transplanted MSCs can integrate into hepatic parenchyma and express hepatocyte markers (ALB, CK18, HNF4α), contributing directly to tissue restoration.
3.2 Paracrine and Anti-Fibrotic Effects
Beyond differentiation, the regenerative effects of MSCs largely depend on paracrine signaling. MSCs secrete a wide array of bioactive molecules—HGF, 血管内皮生长因子 (血管内皮生长因子), insulin-like growth factor-1 (胰岛素样生长因子1), and interleukin-10 (白细胞介素10)—that promote hepatocyte proliferation and suppress inflammation.
HGF in particular plays a pivotal role: it binds to the c-Met receptor on hepatocytes, activating the PI3K/Akt and MAPK/ERK pathways, which drive cell survival and proliferation. Meanwhile, IL-10 and prostaglandin E2 derived from MSCs inhibit TNF-α and TGF-β1 expression in activated stellate cells, limiting fibrogenesis. 因此, MSCs help shift the hepatic microenvironment from a pro-fibrotic to a regenerative state.
3.3 免疫调节
MSCs exert broad immunosuppressive effects that counteract chronic inflammation in cirrhotic tissue. They inhibit T-cell proliferation, induce regulatory T-cells (调节性T细胞), and suppress macrophage M1 polarization while promoting an anti-inflammatory M2 phenotype. Through secretion of indoleamine-2,3-dioxygenase (IDO) and HLA-G, MSCs attenuate immune-mediated injury, providing a more permissive niche for regeneration.
3.4 Anti-Apoptotic and Anti-Oxidative Actions
Oxidative stress is central to hepatocellular apoptosis in cirrhosis. MSC-derived exosomes and microvesicles contain antioxidant enzymes (superoxide dismutase, 过氧化氢酶) 和微小RNA (miR-122, miR-21) that reduce ROS generation and activate survival pathways (例如。, Nrf2-Keap1). These vesicles represent a cell-free therapeutic extension of MSC therapy.
4. Molecular Signaling Pathways
4.1 HGF/c-Met Axis
HGF secreted by MSCs binds to the c-Met receptor, stimulating hepatocyte proliferation and enhancing ECM degradation through up-regulation of matrix metalloproteinases (MMP-2, MMP-9). This pathway is critical for both anti-fibrotic remodeling and hepatocyte regeneration.
4.2 TGF-β/Smad Pathway
In cirrhosis, TGF-β1 is a major driver of fibrogenesis via activation of Smad2/3 signaling in HSCs. MSCs can suppress this pathway by releasing decorin and HGF, which neutralize TGF-β1 activity. 此外, MSC-derived miR-29 family members target collagen type I and III mRNAs, attenuating ECM deposition.
4.3 Wnt/β-Catenin Pathway
Wnt signaling promotes hepatocyte proliferation and regeneration after injury. MSCs secrete Wnt ligands that activate β-catenin translocation in hepatic progenitor cells, enhancing their differentiation. This pathway works synergistically with HGF/c-Met to restore parenchymal mass.
4.4 IL-6/STAT3 Signaling
The IL-6/STAT3 axis mediates acute-phase hepatoprotection and regeneration. MSCs release IL-6 and leukemia inhibitory factor (LIF), which activate STAT3 in hepatocytes, inducing expression of survival genes (Bcl-xL, cyclin D1). STAT3 activation also supports angiogenesis through VEGF up-regulation.
5. Homing and Engraftment of MSCs in the Liver
Once administered systemically (typically via peripheral intravenous infusion), MSCs migrate preferentially to injured tissues through chemotactic gradients. Stromal cell-derived factor-1 (SDF-1), secreted by damaged hepatocytes and endothelial cells, binds to the CXCR4 receptor on MSCs, guiding their homing. Additional adhesion molecules—VCAM-1, ICAM-1, and integrins—facilitate extravasation into hepatic sinusoids.
Within the injured liver, the inflamed microenvironment provides signals such as HGF, 血小板衍生生长因子, and IL-8 that enhance MSC retention and survival. 尤其, even when engraftment efficiency is low (<5%), the paracrine actions of MSCs exert systemic benefits by modulating immune and fibrotic responses.
6. MSC Isolation and Ex Vivo Expansion (Research Context)
In preclinical and clinical studies, autologous MSCs are commonly isolated from bone marrow aspirates 或者 peripheral blood after mobilization. Peripheral blood MSCs can be enriched after administration of granulocyte colony-stimulating factor (G-CSF), which increases circulating progenitor cells.
隔离 typically involves density-gradient centrifugation (例如。, Ficoll-Paque) to separate mononuclear cells, followed by adherence culture in flasks using low-glucose Dulbecco’s Modified Eagle Medium (DMEM) supplemented with fetal bovine serum (胎牛血清). Non-adherent cells are removed after 24–48 hours.
Cultivation and expansion proceed over several passages, generally lasting 7–10 days per cycle, until sufficient cell numbers are reached (often tens of millions of MSCs per infusion in reported trials). Cells are characterized by spindle-shaped morphology and specific surface markers—positive for CD73, CD90, CD105, and negative for CD34, CD45, CD14.
Before infusion, cells are typically washed, resuspended in physiological medium, and subjected to quality control tests for viability and sterility. These procedures, as reported in clinical studies conducted in Europe and Asia, aim to ensure autologous cell purity and minimize immunologic risks.
7. Clinical Outcomes and Challenges
Several clinical trials have assessed autologous MSC infusion in patients with liver cirrhosis of various etiologies (viral, alcoholic, or cryptogenic). Reported benefits include:
- Improvement in serum albumin levels and prothrombin time,
- Reduction in serum bilirubin and transaminases,
- Decrease in MELD (Model for End-Stage Liver Disease) score,
- Enhanced patient quality of life.
Histological studies reveal partial reversal of fibrosis and increased hepatocyte proliferation. 然而, responses vary depending on disease stage, MSC source, and dose.
挑战 remain in ensuring long-term engraftment and functional integration. Repeated infusions may be necessary due to limited persistence of transplanted cells. There is also ongoing debate about the relative contribution of differentiation versus paracrine effects. 而且, the microenvironment of advanced cirrhosis (缺氧, 氧化应激, and matrix stiffness) may restrict MSC survival.
Emerging strategies include preconditioning MSCs with hypoxia or cytokines to enhance their regenerative potency, 和 genetic modification to overexpress HGF or anti-fibrotic factors. 使用 MSC-derived exosomes is another promising cell-free alternative that may replicate therapeutic effects with improved safety and scalability.
8. 未来的方向
Advances in biomaterials and tissue engineering could further improve MSC therapy outcomes. Three-dimensional (3D) culture systems and bioreactors allow large-scale expansion of MSCs while preserving their stemness. Combining MSCs with bioengineered scaffolds or hydrogels may enhance hepatic engraftment and paracrine signaling.
而且, the integration of omics technologies—transcriptomics, proteomics, and metabolomics—will refine our understanding of MSC-liver interactions and identify biomarkers predicting therapeutic response. Regulatory standardization remains essential to translate these therapies into routine clinical practice safely.
9. 结论
Mesenchymal stem cell therapy represents a paradigm shift in the management of liver cirrhosis. Through a multifaceted mechanism involving differentiation, 旁分泌信号传导, 免疫调节, and antifibrotic actions, autologous MSCs can partially restore hepatic structure and function. Molecularly, pathways such as HGF/c-Met, Wnt/β-catenin, and IL-6/STAT3 orchestrate hepatocyte proliferation and tissue remodeling, while suppression of TGF-β/Smad signaling mitigates fibrosis.
参考 (selected)
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- Kharaziha, 磷. 等人. (2009). Improvement of liver function in liver cirrhosis patients after autologous mesenchymal stem cell injection: a clinical study. European Journal of Gastroenterology & 肝病学, 21(10), 1199–1205.
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- Wang, Y., Lian, F., Li, J. 等人. (2012). Adipose-derived mesenchymal stem cells promote liver regeneration and suppress liver fibrosis in a rat model. Journal of Cellular and Molecular Medicine, 16(4), 918–929.*
