ABSTRACT
ObjectiveTo observe the effect of Huanglian Jiedutang on the inflammatory injury in the mouse model of acute gouty arthritis （AGA） and to explore the mechanism of Huanglian Jiedutang in treating AGA. MethodForty male C57BL/6J mice were randomized into blank， model， colchicine （0.83 mg·kg-1）， and Huanglian Jiedutang （5 g·kg-1） groups. The mouse model of AGA was established by injecting monosodium urate （MSU） crystals into the ankle joint. The swelling degree of the right ankle joint of each mouse was measured every day for 7 days， and the pathological changes of the ankle joint were detected by hematoxylin-eosin （HE） staining after 7 days. The other 40 C57BL/6J mice were grouped as above. After 18 hours of modeling， the right ankle joint was collected， and real-time polymerase chain reaction was employed to measure the mRNA levels of interleukin （IL）-1β， tumor necrosis factor （TNF）-α， IL-6， NOD-like receptor protein 3 （NLRP3）， apoptosis-associated speck-like protein containing a CARD （ASC）， and Caspase-1. The expression levels of IL-1β， TNF-α， and IL-6 were measured by the enzyme-linked immunosorbent assay. Western blot was employed to determine the protein levels of NLRP3 inflammasome， Toll-like receptor 4 （TLR4）， and nuclear factor-κB （NF-κB）. ResultCompared with the blank group， the model group showed swelling right ankle joint （P<0.01）， obvious foreign body granuloma in the ankle joint with inflammatory cell infiltration. After the treatment with Huanglian Jiedutang， the ankle joint swelling was relieved （P<0.05， P<0.01）， and the size of foreign body granuloma was reduced. Compared with the blank group， the model group showed up-regulated mRNA levels of IL-1β， TNF-α， and IL-6 in the ankle joint tissue （P<0.01）， up-regulated mRNA levels of NLRP3 and Caspase-1 in the NLRP3 inflammasome （P<0.05， P<0.01）， and up-regulated protein levels of NLRP3， Caspase-1， TLR4， and NF-κB （P<0.05， P<0.01）. Huanglian Jiedutang down-regulated the mRNA levels of IL-1β， TNF-α， IL-6， NLRP3， and Caspase-1 （P<0.05， P<0.01） and the protein levels of IL-1β， TNF-α， IL-6， NLRP3， Caspase-1， TLR4， and NF-κB （P<0.05 or P<0.01）. ConclusionInjecting MSU crystal resulted in local inflammatory injury of the joints in the mouse model of AGA. The treatment with Huanglian Jiedutang may alleviate the inflammatory injury by regulating the NLRP3 inflammasome and TLR4/NF-κB signaling pathway.
ABSTRACT
ObjectiveTo observe the effects of Huanglian Jiedutang on pathological and immune damage in collagen-induced arthritis （CIA） model mice， and to explore the possible mechanism of Huanglian Jiedutang in relieving rheumatoid arthritis. MethodTwenty-four DBA/1 mice were randomly divided into normal group， model group， methotrexate group and Huanglian Jiedutang group， with six mice in each group. The CIA mice model were established using type Ⅱ collagen induction. The administration groups were respectively treated with Huanglian Jiedutang （5 g·kg-1） and methotrexate （0.5 mg·kg-1）. The joint swelling symptoms of the mice were observed， and the arthritis index was scored every 3 days. Flow cytometry was employed to detect granulocytes， monocytes， and T lymphocytes in peripheral blood. The expression of inflammatory cytokines in joint was determined by real-time polymerase chain reaction （Real-time PCR）. The ankle joint was scanned by micro-computed tomography （Micro-CT）， and the histopathological changes were observed through hematoxylin-eosin （HE） staining. ResultCompared with the normal group， the modeling led to joint swelling， elevated the joint index score （P<0.05）， increased the proportion of granulocytes （P<0.05） and decreased that of monocytes and T lymphocytes （P<0.01） in peripheral blood， and raised the neutrophil-to-lymphocyte ratio （NLR） （P<0.01）. Further， it up-regulated the expression of inflammatory cytokines such as tumor necrosis factor-α （TNF-α）， interleukin （IL）-1β， and IL-6 in joint （P<0.01）. Micro-CT showed obvious bone destruction in the ankle joint， and pathological examination revealed the infiltration of a large number of inflammatory cells and the synovial hyperplasia of joint tissue. Compared with the model group， Huanglian Jiedutang alleviated the symptoms of joint swelling， lowered the score of arthritis index （P<0.05）， increased the proportion of T lymphocytes and lowered NLR （P<0.01）. Moreover， it down-regulated the expression levels of TNF-α， IL-1β， and IL-6 in joint （P<0.01） and alleviated the bone destruction and pathological injury of joint tissue. ConclusionType Ⅱ collagen caused systemic and local inflammatory immune damage in CIA mice. Huanglian Jiedutang alleviates such injury， especially for that in local joint， thereby inhibiting joint injury and bone destruction in CIA mice.