Effect of Puerarin, Baicalin and Berberine Hydrochloride on the Regulation of IPEC-J2 Cells Infected with Enterotoxigenic Escherichia coli.

Xiaoxi Liu,Jianqin Xu,Yunfei Ma,Fenghua Liu,Huanrong Li,Xianghong Ju

doi:10.1155/2019/7438593

Abstract

Puerarin, baicalin and berberine hydrochloride are the main components of Gegen Qinlian Decoction, which has been used to treat diarrhoea in China for hundreds of years, yet the biological function and molecular mechanism of these components are not clear. To investigate the effects of puerarin, baicalin, and berberine hydrochloride on the regulation of porcine intestinal epithelial cells (IPEC-J2 cells) infected with enterotoxigenic Escherichia coli (ETEC). IPEC-J2 cells were pretreated with puerarin (200 μg/mL), baicalin (1 μg/mL), and berberine hydrochloride (100 μg/mL) at 37°C for 3 h and then coincubated with the F4ac ETEC bacterial strain 200 at 37°C for 3 h. ETEC infection damaged the structure of IPEC-J2 cells, upregulated mucin 4 (P < 0.01) and mucin 13 mRNA (P < 0.05) expression, increased the apoptosis rate (P < 0.05), and promoted inflammatory responses (IL-6 and CXCL-2 mRNA expression) in IPEC-J2 cells by activating the nuclear factor-κB (NF-κB) signaling pathway. Pretreatment with puerarin, baicalin, and berberine hydrochloride improved the structure and morphology of IPEC-J2 cells and inhibited ETEC adhesion by downregulating specific adhesion molecules. Pretreatment with baicalin decreased the inflammatory response; pretreatment with baicalin and berberine hydrochloride decreased the inflammatory response mediated by the NF-κB signaling pathway. Pretreatment with puerarin, baicalin, and berberine hydrochloride protected IPEC-J2 cells from ETEC infection by inhibiting bacterial adhesion and inflammatory responses.

Highlights

As an important etiological factor of postweaning diarrhoea, enterotoxigenic Escherichia coli (ETEC) induces severe diarrhea in postweaned piglets and causes great economic loss to the swine industry [1, 2]
The pharmacokinetics of Gegen Qinlian Decoction after oral administration to rats showed the concentration of puerarin is the highest in the bioactive isoflavonoids of Puerariae Radix, as well as baicalin is Scutellariae Radix and berberine in Coptidis Rhizoma [19]
Because of their high concentrations and potential efficacy in Gegen Qinlian Decoction, puerarin, baicalin, and berberine hydrochloride were selected to coculture with IPEC-J2 cells in vitro

Summary

Introduction

As an important etiological factor of postweaning diarrhoea, enterotoxigenic Escherichia coli (ETEC) induces severe diarrhea in postweaned piglets and causes great economic loss to the swine industry [1, 2]. ETEC adheres to intestinal epithelial cells after binding to specific receptors [4], producing heat-stable enterotoxins (ST, including STa and STb) and heat-labile enterotoxin, which disrupt host cell functions, stimulate fluid and electrolyte secretion, and eventually cause diarrhea [2, 5, 6]. As a striking candidate gene of the F4ac (F4 fimbriae antigenic variant) ETEC receptor, mucin 4 (Muc4) contributes to the immune system as a major component of the glycocalyx layer of epithelial cells [10, 11]. To counter ETEC invasion, the intestinal epithelium activates multiple innate defense mechanisms [13]; microarray clustered terms of differentially expressed genes in porcine intestinal epithelial cells (IPEC-J2)

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