Effect Of Baicalin On Ulcerative Colitis PI3K/Akt/NF-κB Signaling Pathways And Its Underlying Mechanism

[Experiment in vivo]Objective:to establish the ulcerative colitis model of rats induced by TNBS, to ensure the success of the model, given different doses of baicalin after treatment, respectively, with willow nitrogen sulfanilamide pyridine (SASP) as contrast, baicalin on ulcerative colitis model rats group AKT/NF-kB signaling pathways;Methods:Elisa and Western blot technique, observation on the UC rats colon tissue TNF alpha, IL-6 (interleukin 6), IL-8 (interleukin 8), IL-1 (interleukin 1) content;And PI3K protein, AKT protein, the influence of NF-Bκ(p65), and analyze the PI3K/AKT with other observation index (nf-kappa B, TNF alpha, IL-6) the relationship between, observe the anti-inflammatory mechanisms and the correlation of PI3K/AKT pathway.Using Western blot technique, observation of baicalin on UC model rats colon tissue FasL, the influence of Caspase9, and analyze the PI3K/AKT/FasL nf-kappa B and white, the relationship between Caspase9, observe its antiapoptotic mechanisms and PI3K/AKT/the nf-kappa B pathway.Results:compared with model group, baicalin, willow nitrogen sulfanilamide pyridine group rats colon tissue TNF alpha, IL-6 (interleukin 6), IL-8 (interleukin 8), IL-1 (interleukin 1) production is to reduce (P< 0.05), and two medicine combined application of TNF alpha, IL-6, IL-8, the secretion of IL-1 the weakening effect of greater (P< 0.01).Compared with model group, baicalin, willow nitrogen sulfanilamide pyridine group of PI3K protein phosphorylation level decreased, phosphorylated Akt level decreased (P< 0.05).Compared with model group, baicalin, willow nitrogen sulfanilamide pyridine group of Cox-2 (ring oxidase), beta-catenin (beta serial protein), Caspase-9 (9) aspartic acid proteolytic enzyme, Fasl (human factors associated with apoptosis ligand) protein expression reduced (P< 0.05).Conclusion:baicalin can reduce the inflammation of ulcerative colitis in rats, inhibition of PI3K phosphorylation, cut Akt activation, thus inhibiting the TNF alpha, IL-6, such as the secretion of inflammatory cytokines, play to its anti-inflammatory effect, baicalin may through to inhibit the activation of Akt, inhibit the nf-kappa B play a role of the anti-inflammatory nuclear transfer.[Experiment in vitro]Objective:To investigate the effect of baicalin on colon epithelial cell line (HT-29) cell inflammation model of PI3K/Akt/NF-κB signaling pathways. Methods:Using tumor necrosis factor alpha (tumor necrosis factor alpha, TNF-a) and lipopolysaccharide (lipopolysaccharides, LPS) induced HT-29 cell inflammation model. Experiment is divided into seven groups:blank control group contained complete medium, without any drug intervention during experiments; model group incubated with hTNF(20μg·L-1) for 12 h, then incubated with LPS(1 mg·L-1) for 15 h; Positive medicine group on the basis of the model group, incubated with SASP (500μmol·L-1) for 24 h; Baicalin on the basis of the model group, incubated with different doses (1、10、100μg·L-1) incubation for 24 h. MTT was used to detect the effect of baicalin (1、10、100 μg·L-1) on cell growth; Westernblot was used to measure the protein expression of PI3K, Akt and NF-κB; ELISA was used to measure TNF-a, IL-6 and other content in cell supernatant. Results:Compared with model group, the secretion of TNF-a, IL-6, IL-8, and IL-1 in the cell supernatant reduced in baicalin groups(1、10、100μg·L-1)and SASP group (500μmol·L-1) (P<0.05), and the application of combine with two medicine reduces more secretion of TNF-a, IL-6, IL-8, and IL-1 in the cell supernatant (P<0.01). Compared with model group, the PI3K protein phosphorylation level, phosphorylated Akt level and the activation of the NF-κB level of baicalin (1、10、100μg·L-1), and SASP group (500μmol·L-1) were decreased(P< 0.05). Compared with model group, the Cox-2, β-catenin, Caspase-9, Fasl protein expression decreased in baicalin (1、10、100μg·L-1) and SASP group (500μmol·L-1) (P< 0.05). Conclusion:Baicalin can reduce HT-29 cell inflammation, and inhibit the expression of PI3K/Akt/NF-κB signaling pathways, thus inhibited the secretion of TNF-a, IL-6, IL-8, and IL-1, baicalin may through to inhibit the activation of PI3K/Akt/NF-KB signaling pathways to show the role of its Anti-inflammation.