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  • Hyaluronan 35 kDa enhances epithelial barrier function and protects against the development of murine necrotizing enterocolitis.

Hyaluronan 35 kDa enhances epithelial barrier function and protects against the development of murine necrotizing enterocolitis.

Pediatric research (2019-09-10)
Aarthi Gunasekaran, Jeffrey Eckert, Kathryn Burge, Wei Zheng, Zhongxin Yu, Sean Kessler, Carol de la Motte, Hala Chaaban
ABSTRACT

Disruption of tight junctions (TJs) predisposes to bacterial translocation, intestinal inflammation, and necrotizing enterocolitis (NEC). Previously, studies showed that hyaluronan (HA), a glycosaminoglycan in human milk, maintains intestinal permeability, enhances intestinal immunity, and reduces intestinal infections. In this study, we investigated the effects of HA 35 kDa on a NEC-like murine model. Pups were divided into Sham, NEC, NEC+HA 35, and HA 35. Severity of intestinal injury was compared using a modified macroscopic gut scoring and histologic injury grading. The effect of HA 35 on intestinal permeability was determined by measuring FITC dextran and bacterial translocation. RNA and protein expression of TJ proteins (claudin-2, -3, -4, occludin, and ZO-1) were compared between the groups. Pups in the NEC+HA 35 group had increased survival and lower intestinal injury compared to untreated NEC. In addition, HA 35 reduced intestinal permeability, bacterial translocation, and proinflammatory cytokine release. Ileal expression of claudin-2, -3, -4, occludin, and ZO-1 was upregulated in NEC+HA 35 and HA 35 compared to untreated NEC and shams. These findings suggest that HA 35 protects against NEC partly by upregulating intestinal TJs and enhancing intestinal barrier function.

MATERIALS
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Brand
Product Description

Millipore
Protease Inhibitor Cocktail Set III, Animal-Free, Protease Inhibitor Cocktail Set III, Animal-Free, is a cocktail of six protease inhibitors with broad specificity for the inhibition of aspartic, cysteine & serine proteases & aminopeptidases.
Sigma-Aldrich
Phenylmethanesulfonyl fluoride solution, ~0.1 M in ethanol (T)
Millipore
Phosphatase Inhibitor Cocktail Set II, A cocktail of five phosphatase inhibitors for the inhibition of acid and alkaline phosphatases as well as protein tyrosine phosphatases (PTPs). Suitable for use with cell lysates and tissue extracts.