Effects of Huanglong decoction on mRNA and protein expression levels of intestinal mucosal epithelial Claudin-1 and Occludin in sepsis rat / 中国中西医结合急救杂志

ABSTRACT

Objective To investigate the effects of Huanglong decoction on mRNA and protein expression levels of intestinal epithelial tight junction protein Claudin-1 and Occludin in rat with sepsis. Methods Fifty-five Wistar rats were randomly divided into sham operation group (n=10), model group (n=15), western medicine treatment group (15 rats) and combined traditional Chinese and western medicine group (n=15). In sepsis model group, cecal ligation with puncture (CLP) was reproduced. In the sham operation group, the steps of operation performed were similar to those in the model group, but without CLP. Normal saline 2 mL was given to sham operation group and model group by gavage;meropenem 50 mg was given to the western medicine treatment group by intra-peritoneal injection; in the combined traditional Chinese and western medicine treatment group, besides meropenem 50 mg was injected intra-peritoneally, 2 mL Huanglong decoction (containing raw herbal medicine 0.68 g/mL) was administered by gavage [ingredients of the decoction: rhubarb 15 g (put into water the latest), Glauber salt 10 g, fructus aurantii immaturus 15 g, mangnoliae officinalis 10 g, ginseng 10 g, Angelica sinensis 20 g, Scutellaria baicalensis 15 g, Radix Glycyrrhizae 10 g, concentrated to a decoction 150 mL], in the four groups, the respective drugs or normal saline were given twice a day. After 4 days, the rats were sacrificed, and a partial ileal tissue close to the ileocecal region about 10 cm in length was taken; and mRNA expression levels of Claudin-1 and Occludin were detected by reverse transcription-polymerase chain reaction (RT-PCR), and protein expression levels of Claudin-1 and Occludin were detected by Western Blot. Results Compared with the sham operation group, the expression levels of Claudin-1 and Occludin mRNA [integrated absorbance (IA) ratio] and protein (gray value ratio) were significantly lowered in model group, western medicine group and combined therapy group (Claudin-1 mRNA: 0.327±0.029, 0.387±0.035, 0.420±0.053 vs. 0.561±0.035, Occludin mRNA: 0.364±0.058, 0.422±0.033, 0.442±0.048 vs. 0.616±0.036, Claudin-1 protein: 0.307±0.015, 0.337±0.017, 0.355±0.027 vs. 0.426±0.017, Occludin protein: 0.326±0.028, 0.355±0.017, 0.366±0.025 vs. 0.455±0.018, P<0.05 or P<0.01). Compared with the model group, the expression levels of Claudin-1 and Occludin mRNA and protein in western medicine group and combined therapy group were significantly higher (P < 0.05 or P < 0.01); the up-regulation was more marked in combined therapy group (all P < 0.05). Conclusion The protective effect of Huanglong decoction on intestinal mucosal barrier in rats with sepsis is possibly related to the up-regulation of expression levels of Claudin-1 and Occludin mRNA and protein.