Objective To explore the effect of San Ren decoction on macrophage polarization in Helicobacter pylori(Hp)-associated gastritis(HAG)with spleen and stomach damp heat syndrome through Tim-3/Galectin-9 signaling pathway.Methods Seventy-two Wistar rats were randomly divided into control group,model group,quadruple therapy group(omeprazole 2 mg/kg+amoxicillin 100 mg/kg+clarithromycin 50 mg/kg+colloidal pectin secret capsule 35 mg/kg),San Ren decoction low dose group(3 g/kg),San Ren decoction medium dose group(7.5 g/kg),San Ren decoction high dose group(15 g/kg),12 animals in each group.The rat model of HAG spleen and stomach damp heat syndrome was constructed by composite factors(fatigue+diet+bitter cold medicine+environment+biological factors(Hp bacterial solution)),and the drugs were continuously administered for 21 days according to the groups.The basic conditions of rats were observed in the experiment.Hp colonization was detected by rapid urease test.And gastric mucosal inflammation was observed using HE staining.A transmission electron microscope was used to observe the ultrastructure of the gastric mucosa.The levels of IFN-γ,TNF-α,IL-4,IL-10 in serum were measured by ELISA.Gastric mucosa expressed CD68 by immunohistochemistry.Subtypes of macrophages F4/80+CD86+and F4/80+CD206+were detected by flow cytometry.RT-qPCR was performed to detect the mRNA expression of MCP-1,CD80,CD86,IL-4,CD206,and CD163 in gastric mucosa.Western blot was used to detect the expression of Tim-3 and Galectin-9 proteins in gastric mucosal tissues.Results The histopathology of gastric mucosa in the model group was significantly damaged compared with that of the control group,and the rate of Hp colonization,serum IFN-γ and TNF-α,protein expression of CD68,Tim-3 and Galectin-9 in gastric mucosa tissues,and mRNA expression of MCP-1,CD80,CD86,and CD163 were significantly higher compared with that of the control group(P<0.01).The high-dose intervention of San Ren decoction significantly improved the gastric mucosal tissue injury in model rats,significantly reduced the rate of Hp colonization,and inhibited serum IFN-γ and TNF-α,gastric mucosal tissue CD68,Tim-3,and Galectin-9 protein expression,gastric mucosal tissue MCP-1,CD80,and CD86,CD163 mRNA expression(P<0.05).Conclusion San Ren decoction can improve the symptoms,inhibit the inflammatory response and mediate the macrophages polarization of gastric mucosal in rats with HAG spleen and stomach damp heat syndrome,and its mechanism of action might be related to the regulation of Tim-3/Galectin-9 pathway.

Objective To explore the effect of San Ren decoction on macrophage polarization in Helicobacter pylori(Hp)-associated gastritis(HAG)with spleen and stomach damp heat syndrome through Tim-3/Galectin-9 signaling pathway.Methods Seventy-two Wistar rats were randomly divided into control group,model group,quadruple therapy group(omeprazole 2 mg/kg+amoxicillin 100 mg/kg+clarithromycin 50 mg/kg+colloidal pectin secret capsule 35 mg/kg),San Ren decoction low dose group(3 g/kg),San Ren decoction medium dose group(7.5 g/kg),San Ren decoction high dose group(15 g/kg),12 animals in each group.The rat model of HAG spleen and stomach damp heat syndrome was constructed by composite factors(fatigue+diet+bitter cold medicine+environment+biological factors(Hp bacterial solution)),and the drugs were continuously administered for 21 days according to the groups.The basic conditions of rats were observed in the experiment.Hp colonization was detected by rapid urease test.And gastric mucosal inflammation was observed using HE staining.A transmission electron microscope was used to observe the ultrastructure of the gastric mucosa.The levels of IFN-γ,TNF-α,IL-4,IL-10 in serum were measured by ELISA.Gastric mucosa expressed CD68 by immunohistochemistry.Subtypes of macrophages F4/80+CD86+and F4/80+CD206+were detected by flow cytometry.RT-qPCR was performed to detect the mRNA expression of MCP-1,CD80,CD86,IL-4,CD206,and CD163 in gastric mucosa.Western blot was used to detect the expression of Tim-3 and Galectin-9 proteins in gastric mucosal tissues.Results The histopathology of gastric mucosa in the model group was significantly damaged compared with that of the control group,and the rate of Hp colonization,serum IFN-γ and TNF-α,protein expression of CD68,Tim-3 and Galectin-9 in gastric mucosa tissues,and mRNA expression of MCP-1,CD80,CD86,and CD163 were significantly higher compared with that of the control group(P<0.01).The high-dose intervention of San Ren decoction significantly improved the gastric mucosal tissue injury in model rats,significantly reduced the rate of Hp colonization,and inhibited serum IFN-γ and TNF-α,gastric mucosal tissue CD68,Tim-3,and Galectin-9 protein expression,gastric mucosal tissue MCP-1,CD80,and CD86,CD163 mRNA expression(P<0.05).Conclusion San Ren decoction can improve the symptoms,inhibit the inflammatory response and mediate the macrophages polarization of gastric mucosal in rats with HAG spleen and stomach damp heat syndrome,and its mechanism of action might be related to the regulation of Tim-3/Galectin-9 pathway.

OBJECTIVE To investigate the effects of Sanren decoction on digestion and absorption function and Akt/NF-κB pathway in rats with Helicobacter pylori(Hp)-associated gastritis(HAG) and spleen and stomach damp heat syndrome. METHODS Seventy-two Wistar rats were randomly divided into blank group(n=12) and modeling group(n=60), and a rat model of HAG with combination of factors[fatigue+diet+bitter cold medicine+environment+biological factors(Hp bacterial solution)] was constructed. After successful modeling, the rats were randomly divided into model group, quadruple therapy group(omeprazole 2 mg·kg−1+amoxicillin 100 mg·kg−1+clarithromycin 50 mg·kg−1+colloidal pectin secret capsule 35 mg·kg−1), Sanren decoction low, medium, high dose groups(3, 7.5, 15 g·kg−1), each group was continuously administered for 21 d. During the experiment, the general conditions of rats were observed; the rapid urease test was performed to detect Hp colonization rate; HE staining was performed to observe the inflammation of gastric mucosa; TUNEL was performed to detect apoptosis in gastric mucosa; the content of Ghrelin, IFN-γ, MTL, GAS, IL-4, IL-10 in serum were detected by ELISA; Western blotting was used to detect the protein expression of caspase-3, Bax, Bcl-2, cAMP, Akt, p-Akt, NF-κB p65, p-NF-κB p65 in gastric mucosa; Real-time PCR was performed to detect Akt, NF-κB p65 mRNA expression in gastric mucosal tissues. RESULTS Compared with the blank group, the mucosal tissue of rats in the model group was significantly damaged, and the Hp colonization rate, apoptosis, protein expression of caspase-3, Bax, p-Akt, p-NF-κB p65 in gastric mucosal tissue, mRNA expression of Akt, NF-κB p65 in gastric mucosal tissue and serum IFN-γ content were significantly increased, while protein expression of Bcl-2 in gastric mucosal tissue and serum Ghrelin, MTL, GAS, IL-4 and IL-10 levels were significantly decreased(P<0.01). Compared with the model group, the mucosal tissue injury of rats in Sanren decoction high dose group was significantly improved, the Hp colonization rate, apoptosis, protein expression of caspase-3, Bax, p-Akt, p-NF-κB p65 in gastric mucosal tissues, mRNA expression of Akt and NF-κB p65 in gastric mucosal tissues and serum IFN-γ content were significantly reduced, while the protein expression of Bcl-2 in gastric mucosal tissues and serum levels of Ghrelin, MTL, GAS, IL-4 and IL-10 were significantly increased(P<0.05). CONCLUSION Sanren decoction can improve the symptoms, inhibit the inflammatory response and functional dyspepsia in rats with HAG spleen and stomach damp heat syndrome, and the mechanism of action maybe related to the regulation of Akt/NF-κB pathway.