Alterations of metagenomics and metaproteomics associate kidney disease in a combination of opisthorchiasis and nonalcoholic fatty liver disease

ABSTRACT Background Non–alcoholic fatty liver disease (NAFLD) is prevalent worldwide and is associated with chronic kidney disease (CKD). Opisthorchis viverrini (Ov) infection and consumption of high- fat and high-fructose (HFF) diets exacerbate NAFLD leading to nonalcoholic steatohepatitis. Here, we aimed to investigate the effects of a combination of HFF diets and O.viverrini infection on kidney pathology via changes in the gut microbiome and host proteome in hamsters. Methodology/Principal findings Twenty animals were divided into four groups; Normal diet feeding and non-infected Ov (Normal); HFF diets feeding (HFF); Ov infection (Ov); and feeding with a combination of HFF diets and infection with Ov (HFFOv). Fecal samples were extracted and used for Illumina Miseq sequencing platform based on the V3–V4 region of the 16S rRNA gene, along with LC/MS-MS analysis. Histopathological studies and biochemical assays were also conducted. The results indicated that the HFFOv group exhibited the most severe kidney injury, as elevated KIM-1 expression and accumulation of fibrosis in kidney tissue. In comparison with the HFF group, the combined group showed higher diversity and composition. An increased number of Ruminococaceae , Lachospiraceae , Desulfovibrionaceae and Akkermansiaceae , and a lower number of Eggerthellaceae were observed. A total of 243 significant host proteome were identified in all groups. Analysis using STITCH predicted that host proteome associated leaky gut such as soluble CD14 and p-cresol may play a role in the development of kidney disease. Among host proteome, TGF-beta, involving in fibrogenesis, was significantly expressed in HFFOv. Conclusions/Significance The combination of HFF diets and O.viverrini infection may promote kidney injury through the alterations in the gut microbiome and host proteome. This knowledge may be an effective strategy to prevent the progression of CKD beyond the early stages. Author summary A diets high in fat and fructose causes nonalcoholic fatty liver disease (NAFD), which is increasing worldwide. Liver fluke ( Opisthorchis viverrini , Ov) infection is endemic in the Mekong subregion including in the northeastern Thailand. The prevalence of opisthorchiasis caused by the infection with O. viverrini is associated with fatty liver and bile duct cancer. We have previously demonstrated that infection with O. viverrini exacerbates NAFD progression to non-alcoholic steatohepatitis (NASH) in animal models. NASH exists kidney disease severity higher than ingestion of high-fat and high-fructose (HFF) diets or infection with O. viverrini . Here, we further investigate whether metagenomics is more likely to change in NASH than in NAFD or opisthorchiasis conditions. The combined group had higher diversity and composition. Elevated levels of Ruminococaceae , Lachospiraceae , Desulfovibrionaceae and Akkermansiaceae and decreased levels of Eggerthellaceae were observed, suggesting that HFF+Ov may cause gut dysbiosis in NASH. Differentially expressed proteins were also associated with these gut microbiomes in NASH condition. In addition, we found that the association of metagenomics and metaproteomics in NASH was related to kidney disease. Analysis using STITCH predicted that host proteome may be involved in leaky gut such as soluble CD14 and p-cresol to promote kidney disease. A significantly expressed TGF-beta involving fibrogenesis was found to be associated with kidney fibrosis. Therefore, alterations of metagenomics and metaproteomics is associated with kidney disease in a combination of opisthorchiasis and nonalcoholic fatty liver disease.