Human umbilical cord-derived mesenchymal stem cells ameliorate insulin resistance by suppressing NLRP3 inflammasome-mediated inflammation in type 2 diabetes rats.
BACKGROUND
Insulin resistance is one of the most common and important pathological features of type 2 diabetes (T2D). Recently, insulin resistance is increasingly considered to be associated with systemic chronic inflammation. Elevated levels of tumor necrosis factor (TNF)-α and interleukin (IL)-1β in blood are predictive indicators of the development of T2D. Mesenchymal stem cell (MSC)-based therapies have been proven to have potential immunomodulation and anti-inflammatory properties through their paracrine effects; however, the mechanism for the anti-inflammatory effect of MSCs in enhancing insulin sensitivity is still uncertain.


METHODS
In the present experiment, we used HepG2 cells, a human hepatoma cell line, and a MSC-HepG2 transwell culturing system to investigate the anti-inflammatory mechanism of human umbilical cord-derived MSCs (UC-MSCs) under palmitic acid (PA) and lipopolysaccharide (LPS)-induced insulin resistance in vitro. Insulin resistance was confirmed by glycogen assay kit and glucose assay kit. Inflammatory factor release was detected by ELISA, gene expression was tested by quantitative real-time PCR, and insulin signaling activation was determined by western blotting analysis. The changes of inflammatory factors and insulin signaling protein were also tested in T2D rats injected with UC-MSCs.


RESULTS
Treating HepG2 cells with PA-LPS caused NLRP3 inflammation activation, including overexpression of NLRP3 and caspase-1, and overproduction of IL-1β and IL-18 as well as TNF-α from HepG2 cells. The elevated levels of these inflammatory cytokines impaired insulin receptor action and thereby prevented downstream signaling pathways, exacerbating insulin resistance in HepG2 cells. Importantly, UC-MSCs cocultured with HepG2 could effectively alleviate PA and LPS-induced insulin resistance by blocking the NLRP3 inflammasome activation and inflammatory agents. Furthermore, knockdown of NLRP3 or IL-1β partially improved PA and LPS-induced insulin signaling impairments in the presence of UC-MSCs. Similarly, UC-MSC infusion significantly ameliorated hyperglycemia in T2D rats and decreased inflammatory activity, which resulted in improved insulin sensitivity in insulin target tissues.


CONCLUSIONS
Our results indicated that UC-MSCs could attenuate insulin resistance and this regulation was correlated with their anti-inflammatory activity. Thus, MSCs might become a novel therapeutic strategy for insulin resistance and T2D in the near future.
['Sun|Xiaoya|X|', 'Hao|Haojie|H|', 'Han|Qingwang|Q|', 'Song|Xiaoyan|X|', 'Liu|Jiejie|J|', 'Dong|Liang|L|', 'Han|Weidong|W|http://orcid.org/0000-0002-4415-0874', 'Mu|Yiming|Y|']
D000818:Animals D053148:Caspase 3 D018920:Coculture Techniques D003921:Diabetes Mellitus, Experimental D005260:Female D005312:Fetal Blood D005786:Gene Expression Regulation D056945:Hep G2 Cells D006801:Humans D056747:Immunomodulation D058847:Inflammasomes D007249:Inflammation D007333:Insulin Resistance D020382:Interleukin-18 D053583:Interleukin-1beta D008070:Lipopolysaccharides D008297:Male D045164:Mesenchymal Stem Cell Transplantation D059630:Mesenchymal Stem Cells D000071199:NLR Family, Pyrin Domain-Containing 3 Protein D019308:Palmitic Acid D051381:Rats D017207:Rats, Sprague-Dawley D013311:Streptozocin D014409:Tumor Necrosis Factor-alpha
Mesenchymal stem cells Insulin resistance Inflammation Type 2 diabetes Palmitic acid Lipopolysaccharide NLRP3 inflammasome