目的探讨大黄附子汤含药血清对重症急性胰腺炎小鼠腹腔巨噬细胞功能的影响及其机制。方法制备小鼠重症急性胰腺炎模型,取其腹腔巨噬细胞分成模型组、大黄附子汤含药血清(2.5%、5.0%、10.0%)组、酪氨酸磷酸化抑制剂AG490(10gmol/L)阳性对照组,细胞继续培养2h后,各给药组给予相应含药血清、对照组给予空白血清0.5mL,培养24h,收集上清液。ELISA法检测细胞上清液中肿瘤坏死因子(TNF-α)、白细胞介素-6(IL-6)水平,Westernblotting法检测pJAK2、pSTAT3基因和蛋白表达水平。结果大黄附子汤能显著减少大鼠腹腔巨噬细胞分泌的IL-6、TNF-α,抑制pJAK2、pSTAT3基因及蛋白的表达。结论大黄附子汤含药血清可能通过降低pJAK2、pSTAT3基因和蛋白的表达,阻断JAK2/STAT3信号转导通路,抑制腹腔巨噬细胞分泌IL-6、TNF-α,从而减轻机体过度的炎症反应和重症急性胰腺炎的病情。
Objective To study the effects of serum containing Dahuang Fuzi (Rhubarb and Aconite) Decoction (DFD) on JAK2/STAT3 signal pathway in mice with severe acute pancreatitis (SAP). Methods SAP model in mice was constructed, and then the peritoneal macrophages were vaccinated into the culture plate to set model group, serum containing DFD groups (2.5%, 5%, and 10%), AG490 (10 μmol/L ) positive group, and peritoneal macrophages of normal mice acted as normal control group. After an incubation of 2 h, cells were added with serum containing DFD at different concentration or AG490 0.5 mL, 24 h later, the concentration of TNF-α and IL-6 in supernatant were determined by quantitative sandwich enzyme-linked immunosorbent assay (ELISA) kits, mRNA and protein expression levels of JAK2 and STAT3 in cells were evaluated by Western blotting. Results DFD could significantly decrease the levels of cytokines TNF-α and IL-6 in the supernatants, inhibit the mRNA and protein expression levels of JAK2 and STAT3 in peritoneal macrophages of SAP mice. Conclusion DFD could inhibit the JAK2/STAT3 signal pathway and inflammatory responses in peritoneal macrophages of SAP mice.