Resveratrol improves the therapeutic efficacy of bone marrow-derived mesenchymal stem cells in rats with severe acute pancreatitis

https://doi.org/10.1016/j.intimp.2019.106128Get rights and content

Highlights

Abstract

Objective

Bone marrow-derived mesenchymal stem cells (BMSCs) are effective in the treatment of severe acute pancreatitis (SAP), but their therapeutic effects could still be improved. In order to optimize the clinical application of BMSCs, we adopted the strategy of resveratrol (Res) pretreatment of BMSCs (Res-BMSCs) and applied it to a rat model of sodium taurocholate (NaT)-induced acute pancreatitis.

Methods

SAP was induced by injection of 3% NaT into the pancreatic duct and successful induction of SAP occurred after 12 h. Rats were treated with BMSCs, Res or BMSCs primed with Res at 40 mmol/L, Vandetanib (ZD6474) daily oral dosages of 50 mg/kg vandetanib.

Results

Res stimulated BMSCs to secrete vascular endothelial growth factor A (VEGFA), activated the downstream phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway, and inhibited pancreatic cell apoptosis. In addition, conditioned medium (CM) from Res-BMSCs enhanced the proliferation of human umbilical vein endothelial cells (HUVECs) in vitro, increased resistance to apoptosis and promoted the expression of angiogenesis-related proteins CD31, VEGF and VEGFR2 in pancreatic tissue, but Vandetanib partly abolished these effects by blocking the VEGFA- mediated pathway.

Conclusion

Resveratrol-preprocessed BMSCs can activate the PI3K/AKT signaling pathway in pancreatic cells and HUVECs through paracrine release of VEGFA; thus, achieving the therapeutic effect of resisting apoptosis of pancreatic cells and promoting regeneration of damaged blood vessels. Res pretreatment may be a new strategy to improve the therapeutic effect of BMSCs on SAP.

Introduction

Acute pancreatitis (AP) is one of the most common gastrointestinal disorders with morbidity of 13–45/100,00 worldwide [1]. Approximately 10–20% of AP cases will progress to severe acute pancreatitis (SAP), which is associated with rapid progression, multiple complications and high mortality [2]. Despite our increased understanding of the pathogenesis of SAP and significant developments in its treatment, the mortality rate of SAP is as high as 15–25% [3], [4]. Even with improvements in intensive care treatment during the past few decades, the mortality of SAP has not declined significantly [5]. Therefore, patients with SAP urgently need new and better treatment measures.

Apoptosis of pancreatic cells is common in SAP, apoptosis of pancreatic cells can lead to the release of various pro-inflammatory factors such as interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α). These pro-inflammatory factors can activate Toll-like receptors on the surface of macrophages in the pancreas, forming a vicious cycle of “pro-inflammatory factor-inflammatory pathway, pro-inflammatory factor”, and aggravation of pancreatic tissue injury [6]. Therefore, Inhibition of apoptosis should be of significant benefit in the treatment of SAP.

Severe previous studies have demonstrated that vascular endothelial cells were damaged in the early stage of SAP [7]. Injury of vascular endothelial cell further leads to disturbance of pancreatic micro-circulation, tissue edema and aggravation of inflammatory reaction. Therefore, the repair of injured blood vessels may become a new target for SAP therapy.

Resveratrol (Res; 3,5,4′-trihydroxy-trans-stilbene) is a plant-derived polyphenolic phytoalexin that is in high concentrations in grape peels and rhizomes of giant knotweeds[8]. It has been reported that Res shows beneficial anti-inflammatory, antioxidant, anti-apoptosis, and angiogenesis effects [9], [10].

In recent years, with the rise of stem cell therapy research, many investigators have confirmed that mesenchymal stem cells (MSCs) have a protective effect against a number of diseases, including acute liver failure [11], acute kidney injury [12], and acute lung injury [13]. MSCs can be easily separated with slight suction of bone marrow and adipose tissue and have a variety of functions, including self-renewal, multi-differentiation, immune regulation, paracrine effects and directional migration, all with little to no immunological rejection after transplantation [14]. Therefore, MSCs may be the ideal cells to repair tissue injury caused by a variety of reasons. Previous research have also revealed that MSCs can repair pancreatic injury and exert anti-apoptosis and promote angiogenesis in SAP [15]. Although MSCs can alleviate SAP, when we used MSCs alone, they failed to achieve the full therapeutic effect because severe inflammation in vivo affects their activity. The efficacy of MSC therapy can thus be further improved, and many researchers have suggested the use of small molecule drugs to regulate the function of stem cells and improve their efficacy [16], [17]. Meanwhile Resveratrol can also protect bone marrow MSCs against inflammatory and oxidative damage [18]. Preconditioning of MSCs with Res has been found to be cytoprotective [18]. However, the specific mechanism has been a controversy so far. In our previous work, we have demonstrated that VEGF played an important role in the process of BMSCs therapy for SAP [15], [19].

Recent studies have also shown that VEGF is an important protective factor when SAP occurs [20]. The most functional form of VEGF subtypes is VEGFA [21]. VEGFA can inhibit apoptosis of pancreatic cells and promote the repair of pancreatic damaged blood vessels [20], [22].

In the current study, we investigated the effect of bone marrow-derived mesenchymal stem cells (BMSCs) preconditioned with Res (Res-BMSCs) for the treatment of rats with SAP. Our findings suggest that BMSCs pretreated with Res can promote the regeneration of pancreatic vessels and inhibit the apoptosis of pancreatic cells by promoting paracrine release of vascular endothelial growth factor A (VEGFA) and activating the phosphatidylinositol-4,5bisphosphate 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway.

Section snippets

Cell culture

BMSCs were isolated, cultured, and identified as described in our previous study [23]. Human umbilical vein endothelial cells (HUVECS) were purchased from Shanghai Cell Bank of the Chinese Academy of Sciences and cultured in DMEM-LG complete medium. The cells were detached with 0.05% trypsin/25 mmol/L EDTA solution and passaged upon reaching 80% confluence. BMSCs obtained between the third and fifth passages were used [23].

Preparation of Res-BMSCs

Res (Sigma, St. Louis, MO, USA) was dissolved in dimethyl sulfoxide

Res-BMSCs were more effective than BMSCs in alleviating SAP

SAP rats were injected with BMSCs, Res or Res-BMSCs intravenously after NaT induction. Rats in the SAP group and Res-BMSCs group daily oral dosages of 50 mg/kg vandetanib. Serum amylase and lipase activities were increased after successful induction of SAP, and transplanted Res-BMSCs significantly reduced serum amylase and lipase levels compared with BMSCs, but Vandetanib, an inhibitor of VEGFR2, counteracted some protective effects of Res-BMSCs (Fig. 1A and B). Pancreatic tissue damage was

Discussion

The incidence of AP is gradually increasing, but the underlying pathogenesis is still unclear [32]. Apoptosis in necrotic pancreatic tissue and vascular injury are important factors leading to aggravation of pancreatic injury [15], [33]. These factors may cause About 20% of AP patients will progress to SAP [34]. SAP is often accompanied by multiple organ failure, including the lungs and kidneys, with high mortality and is more intractable to treatment [3]. At present, the treatment strategy for

Conclusion

In this study, we have demonstrated that Res enhances the therapeutic efficacy of BMSCs in rats with SAP. BMSCs primed with Res have a better capacity to exert anti-apoptotic and angiogenic effects during treatment for SAP by secreting VEGFA to activate PI3K/AKT pathway, this study provides a new therapeutic strategy to improve pancreas function for patients with SAP.

CRediT authorship contribution statement

Dalu Liu: Conceptualization, Methodology, Software, Writing - original draft. Guodong Song: Software, Formal analysis, Data curation, Investigation. Zhilong Ma: Software, Formal analysis, Data curation, Investigation. Xiang Geng: Resources, Validation. Yuxiang Dai: Data curation, Validation. Tingsong Yang: Resources. Hongbo Meng: Resources. Jian Gong: Resources. Bo Zhou: Resources. Zhenshun Song: Conceptualization, Software, Writing - review & editing.

Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (No. 81670582)

Declaration of competing interest

All authors declare that they have no conflict of interests.

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