银耳多糖对小鼠淋巴细胞免疫调节作用及PI3K/Akt、NF-κB信号通路的影响

Effect of Tremella fuciformis polysaccharides on lymphocyte immunomodulatory effect and PI3K/Akt and NF-κB signaling pathway

试验旨在探讨银耳多糖(TFP)对小鼠脾淋巴细胞免疫调节作用及磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(Akt)、核因子κB(NF-κB)信号通路的影响。试验从体外分离小鼠脾淋巴细胞,分别设CK组、LMS组、TFP 20 mg/L组、TFP 40 mg/L组、TFP 80 mg/L组和TFP 160 mg/L组。采用MTT法检测脾淋巴细胞的增殖,ELISA法检测脾淋巴细胞细胞因子白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、白细胞介素-12(IL-12)和干扰素-γ(IFN-γ)的分泌,QRT-PCR法检测脾淋巴细胞细胞因子IL-6、IL-10、IL-12和IFN-γmRNA的相对表达量,Western blot法检测PI3K/Akt和NF-κB信号通路相关蛋白表达的影响。结果显示,银耳多糖能够促进脾淋巴细胞的增殖、IL-6、IL-10、IL-12和IFN-γ的分泌及mRNA的表达(P<0.01),降低磷酸化磷脂酰肌醇-3-激酶(P-PI3 K)、磷酸化蛋白激酶B(P-Akt)、κB抑制因子激酶α(IKK-α)、κB抑制因子激酶β(IKK-β)和NF-κB蛋白表达(P<0.01),提高核因子κB抑制因子α(IκB-α)蛋白的表达(P<0.01)。研究表明,银耳多糖对小鼠脾淋巴细胞具有免疫调节作用,其作用机制可通过激活小鼠脾淋巴细胞PI3K/Akt和NF-κB信号通路,发挥免疫调节作用,TFP为80 mg/L时效果较好。

The experiment was to investigate the effect of Tremella fuciformis polysaccharide on immunomodulatory effect of splenic lymphocytes and the signaling pathways of PI3K/Akt and NF-κB in mice.Mouse splenic lymphocytes were isolated in vitro,and divided into CK group,LMS group,TFP 20 mg/L group,TFP 40 mg/L group,TFP 80 mg/L group and TFP 160 mg/L group.The proliferation of splenic lymphocytes was detected by MTT method,the secretion of splenic lymphocyte cytokines IL-6,IL-10,IL-12 and IFN-γwas detected by ELISA,the relative expression of splenic lymphocyte cytokines IL-6,IL-10,IL-12 and IFN-γmRNA was detected by QRT-PCR,and the effect of PI3K/Akt and NF-κB signaling pathway-related proteins was detected by Western blot.The results showed that Tremella polysaccharides could promote the proliferation of spleen lymphocytes,the secretion of cytokines IL-6,IL-10,IL-12,IFN-γand mRNA expression(P<0.01),and reduce the expression of P-PI3K,p-Akt,IKK-α,IKK-βand NF-κB protein(P<0.01),increase the expression of IκB-αprotein(P<0.01).The experiment indicates that Tremella fuciformis polysaccharide has an immunomodulatory effect on mouse splenic lymphocytes,and its mechanism of action can exert an immunomodulatory effect by activating the PI3K/Akt and NF-κB signaling pathways of mouse splenic lymphocytes.