| Milk β-Lg allergy can affect the absorption of high quality milk protein, and it will endanger the health of the body. The recent reports suggested that the intestinal short chain fatty acid(SCFA) metabolism disorder caused by allergy is closely related to the occurrence and develo pment of allergic inflammation. The role of probiotics in regulating the metabolism of short chain fatty acids has been a concern, but it has not been reported in the ease of milk allergy. In this study, we first observed the effect of butyrate and acetic acid on the lymphocyte of allergic mice in vitro and detected the expression changes of cell proliferation and short chain fatty acid specific receptor,then constructed milk allergy mice model to observe anti allergic properties of Lactobacillus acidophilus on intestinal short chain fatty acid metabolism, monitored the expression of fatty acid receptor GPR41 and GPR43 in the spleen and colon tissues of mice and analysed the change of total Ig E, mice serum inflammatory cytokines and allergic symptoms. Our research is to investigate the relationship between the regulation of fatty acid metabolism and activation of intestinal immune signal and the prevention and treatment of inflammation, revealed the regulating metabolism of probiotic to the short fatty acid by relieving allergic from the cell and whole experiment.In this study, we first studyed the effect of SCFA on inflammation in vitro. The effect of SCFA on the proliferation of splenic lymphocytes in mice was detected by MTT assay. Take a final concentration of 1, 2, 5, 10, 50 and 100 m M/L of sodium acetate and sodium butyrate,coincubat ed with spleen lymphocytes of primary mouse spleen lymphocytes 48 h in vitro.The results showed that sodium acetate had no significant effect on the proliferation of lymphoc ytes in spleen, and sodium butyrate can significantly inhibit the spleen lymphocyte abnormal proliferation and transformation(P<0.05) and showed a dose-dependent manner.And then, the effect of SCFA on the secretion of cytokines and antibodies in the sple en lymphocytes of sensitized mice was detected. Take a final concentration of 1, 2, 5, 10, 50 and 100 mM/L of sodium acetate and sodium butyrate,coincubated with spleen lymphocytes of primary mouse spleen lymphocytes 48 h in vitro. Compared to the stimulation of sodium acetate, sodium butyrate on Ig E and inflammatory cytokines(TNF-a, IL-17 and IL-6) has a good inhibitory effect, and significantly promote the Treg type cytokine TGF-β secretion(P<0.05).Finally, the effect of SCFA on the expression of fatty acid G receptor in the spleen cells of sensitized mice was detected. Take a final concentration of 1, 2, 5, 10, 50 and 100 mM/L of sodium acetate and sodium butyrate,coincubated with spleen lymphocytes of primary mouse spleen lymphocytes 48 h in vitro. The Expression of G protein mRNA in spleen cells was detected by RT-PCR. Compared to the stimulation of sodium acetate and different concentrations of sodium butyrate promoted the induced by sensitized lymphocytes GPR41 and GPR43 mRNA expression(P<0.05), and sodium butyrate on receptor GPR43 expression of stimulative effect is better than that of GPR41.Animal experimental part of this study first examined the acidophilus intervention effects on β-Lg allergic mice intestinal SCFA metabolism.BALB/c mice were randomly divided into blank group, induced by sensitized group, thermophilic acidophilic lactobacillus group(group 0.096 g/kg, group 0.96 g/kg, group 9.6 g/kg) and butyrate sodium group(50 mM/L). The metabolic changes of intestinal SCFA in each group were analyzed by gas chromatography. Compared with the control group, the levels of acetic acid in the feces of sensitized mice were significantly increased, the levels of propionate, butyrate, valeric acid were significantly decreased(P<0.05). The Lactobacillus acidophilus can effectively improve the metabolic disorder of intestinal SCFA allergy. Through the analysis of short chain fatty acids in feces of sensitized mice and control mice, the results showed that compared with the blank group, allergy group colo n feces in acetic acid showed a high abundance, but the abundance of other short chain fatty acids(propionate, butyrate, and acid) decreased obviously in the allergic reaction. Among them, especially the decline of butyrate was the most significant. In different concentrations of probiotics, the abundance of acetic acid decreased significantly, and the abundance of other short chain fatty acids increased, butyrate was the most significant(P<0.05).At the same time, the expression of GPR41 and GPR43 in the spleen and colon of mice in each group was detected by RT-PCR. The results showed that the expression of GPR41 and GPR43 in the allergic group was lower than that in the blank group. The expression levels of GPR41 and GPR43 receptors in the spleen and colon of the probiotic group and sodium butyrate group were dose dependent, and the expression of mRNA GPR43 in colon and spleen was higher than that in GPR41.Finally, the relationship between the expression of G receptor and the remission of β-Lg allergy was analyzed in this study.Compared with the normal mice, the total Ig E and the number of inflammatory cells were significantly increased in the serum of the β-Lg sensitized mice. The pathological results showed that there was a significant inflammatory cell infiltration in the colon and lung tissue. The probiotics and sodium butyrate intervention can effectively relieve the β-Lg allergies, high doses of probiotics group(9.6 g/kg) and sodium butyrate group mice alveolar wall without bleeding, colon adenocarcinoma cells arranged in neat rows, the serum total Ig E levels decreased significantly(P< 0.05), TGF-β and IFN-γ levels significantly up-regulated(P< 0.05). |
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