通络蠲痹颗粒调控ROS-NLRP3-Caspase-1轴对骨关节炎软骨细胞焦亡的影响OA

Objective To explore the effects of Tongluo Juanbi Granules(通络蠲痹颗粒,TLJB)medicated serum on pyroptosis and the reactive oxygen species(ROS)-NOD-like receptor protein 3(NLRP3)-Caspase-1 signaling axis in rabbit chondrocytes at the cellu-lar level,and to study its mechanism of action in preventing and treating osteoarthritis(OA).Methods TLJB medicated serum and blank serum were extracted.Rabbit chondrocytes were cultured in vitro,and divided into normal control group(group A),model group(group B),blank serum group(group C),TLJB medicated serum group(group D),NLRP3 activator group(group E),and TLJB medi-cated serum+NLRP3 activator group(group F).Chondrocytes were identified using Toluidine Blue staining and Type II collagen immu-nofluorescence.MTT assay was used to select the optimal intervention concentration of TLJB medicated serum.Lactate dehydrogenase(LDH)levels were detected by a coupled enzyme assay.Levels of IL-1β and IL-18 were detected by enzyme-linked immunosorbent assay(ELISA).ROS levels were detected by a chemiluminescence method.Real-time quantitative PCR(RT-qPCR)was used to detect the mRNA expression of NLRP3,Caspase-1,GSDMD,IL-1β and IL-18 in chondrocytes.Western blot(WB)was used to detect the protein expression of NLRP3,pro-Caspase-1 and GSDMD.Results MTT assay showed that 15%TLJB medicated serum intervention for 48 hours was optimal for rabbit chondrocyte proliferation.Compared with group A,LDH,IL-1β,IL-18,ROS levels,and mRNA and protein expression of NLRP3,Caspase-1,and GSDMD in group B were significantly increased(P<0.01).Compared with group B,group E showed significant increases in LDH,IL-1β,IL-18,ROS levels,and mRNA and protein expression of NLRP3,Caspase-1,and GSDMD(P<0.01),while group D showed significant decreases in these indicators(P<0.01).Compared with group E,LDH,IL-1β,IL-18,ROS levels,and mRNA and protein expression of NLRP3,Caspase-1,and GSDMD in group F were significantly decreased(P<0.01).Conclusion TLJB medicated serum can inhibit pyroptosis in rabbit articular chondrocytes,and its mechanism may be related to the suppression of the ROS-NLRP3-Caspase1 signaling axis activation.