Effect of Modified Simiaosan on miR-223-3p and NLRP3/IL-1β Signaling Pathway in Rats with Acute Gouty Arthritis / 中国实验方剂学杂志

ABSTRACT

ObjectiveTo investigate the effect of modified Simiaosan on miR-223-3p and NOD-like receptor thermal protein domain associated protein 3 （NLRP3）/interleukin-1β （IL-1β） signaling pathway in rat model with acute gouty arthritis （AGA） and explore the anti-inflammatory mechanism of modified Simiaosan on AGA. MethodA total of 72 8-week-old male SD rats were selected. They were divided into blank group， model group， colchicine group （0.3 mg·kg-1）， high-dose modified Simiaosan group （31.75 g·kg-1）， medium-dose modified Simiaosan group （15.75 g·kg-1）， and low-dose modified Simiaosan group （7.875 g·kg-1） according to random number table method， with 12 rats in each group. Except for the blank group， monosodium urate （MSU） crystal suspension was injected into the right ankle joint of rats by the Coderre method in other groups to replicate the rat model with AGA. The drug administration groups were given the corresponding drug solution by gavage， and the model group and the blank group were given an equal volume of sterile sodium chloride solution by gavage for one week. The circumference of the rats' ankle joint was measured， and the swelling degree of the ankle joint was calculated. Hematoxylin-eosin （HE） staining was used to detect the pathological morphological changes in the synovial tissue of the ankle joint. The levels of IL-1β， tumor necrosis factor-α （TNF-α）， and interleukin-6 （IL-6） in the serum of rats in each group were detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the protein expressions of NLRP3， Caspase-1， and apoptosis-related spot-like protein （ASC） in synovial tissue of rats in each group， and real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expressions of NLRP3， Caspase-1， and ASC and the expression of miR-223-3p in synovial tissue of rats. ResultCompared with that in the normal group， the swelling degree of the ankle joint in the model group was higher （P<0.01）， and the synovial tissue structure was disordered. Synovial cells proliferated obviously， and a large number of inflammatory cells were infiltrated. The levels of IL-1β， IL-6， and TNF-α in the serum of the model group increased significantly （P<0.01）， and the protein and mRNA expressions of NLRP3， Caspase-1， and ASC increased， while expression of miR-223-3 decreased. Compared with the model group， the swelling degree of ankle joint in the colchicine group and high-dose and medium-dose modified Simiaosan groups was lower （P<0.05）. Synovial cell proliferation and inflammatory cell infiltration of the colchicine group and high-dose， medium-dose， and low-dose modified Simiaosan groups were reduced to varying degrees， among which the colchicine group and high-dose modified Simiaosan group improved most obviously. The levels of IL-1β， IL-6， and TNF-α in the serum of rats in different dose groups of modified Simiaosan and colchicine group decreased significantly （P<0.01）， while the protein and mRNA expressions of NLRP3， Caspase-1， and ASC increased （P<0.01）. The expression of miR-223-3p in synovial tissue of the medium-dose and high-dose modified Simiaosan groups and colchicine group increased significantly （P<0.01）. Compared with those in the colchicine group， the levels of IL-1β， IL-6， and TNF-α in the low-dose modified Simiaosan group increased greatly （P<0.01）. In the medium-dose modified Simiaosan group， the protein expressions of NLRP3， Caspase-1， and ASC increased， and the expression of miR-223-3p decreased （P<0.05）. In the low-dose modified Simiaosan group， the levels of IL-1β， IL-6， and TNF-α increased greatly （P<0.01）， as well as the protein and mRNA expressions of NLRP3， Caspase-1， and ASC， while the expression of miR-223-3p was decreased （P<0.01）. ConclusionModified Simiaosan may play an anti-inflammatory role by intervening in the NLRP3/IL-1β signaling pathway via regulating miR-223-3p.