Experimental Study of Zhiqiao Gancao Decoction in Regulating Inflammatory Bodies to Improve Pain Sensitivity of Lumbar Intervertebral Disc Herniation
Objective:To elucidate the activation of Nod-like receptor thermal protein domain associated protein 3(NLRP3)inflammatory corpuscles in nucleus pulposus tissue on the activation of pain sensitive links mediated by acid-sensing ion channel 3(ASIC3),and to explore the intervention mechanism of Zhiqiao Gancao decoction in improving the pain of lum-bar disc herniation(LDH).Methods:60 SD rats were randomly divided into blank group,model group,Caspase-1 inhibitor group,low dose group of Zhiqiao Gancao decoction(crude drug 3 g/kg),medium dose group of Zhiqiao Gancao decoction(crude drug 6 g/kg),and high dose group of Zhiqiao Gancao decoction(crude drug 12 g/kg),with 10 rats in each group.Except for blank group,the LDH model was established in all other groups of rats.5 d after modeling,the rats in each group were given corresponding intervention and intragastric administration for 20 d.The threshold of mechanical and cold stimulation foot shrinkage contraction of rats in each group was measured before modeling and 5,12 and 19 d after model-ing.After extraction,hematoxylin-eosin(HE)staining was used to detect the degree of intervertebral disc degeneration;immunofluorescence was used to detect the expression of Caspase-1p10 in nucleus pulposus tissue;ELISA was used to detect IL-6,TNF-α and IL-1β content;Western Blot was used to detect the expression of NLRP3,Caspase-1p10 in inter-vertebral disc tissue and ASIC3 in dorsal root ganglia(DRG).The real-time polymerase chain reaction(RT-PCR)was used to detect the mRNA expression of ASIC3 in DRG.Results:Mechanical and cold stimulation foot shrinkage threshold results:The LDH model group rats showed typical cold and mechanical pain sensitivity states,and the pain sensitivity state of the rats in the inhibitor group and Zhiqiao Gancao decoction treatment groups were relieved,and the cold stimula-tion and mechanical stimulation foot shrinkage threshold were significantly improved compared with the model group(P＜0.05).HE staining results:In the LDH model group,the fibrous rings were arranged in disorder,and some of them were swollen and broken.Compared with the model group,the arrangement of fibrous rings in the inhibitor group and the Zhiqiao Gancao decoction treatment groups were more orderly,and the fibrous rings were not significantly broken.Immu-nohistochemistry:Compared with the model group,the average optical density of Caspase-1p10 in the treatment group of Zhiqiao Gancao decoction was weaker,with a significant difference(P＜0.05).ELISA results:The inflammatory factors IL-6,IL-1β and TNF-α in the treatment groups decreased significantly compared with model group,with statistical differ-ences(P＜0.05).PCR results:Compared with the model group,the expression of ASIC3 mRNA in the inhibitor group and the Zhiqiao Gancao decoction treatment groups decreased significantly,with statistical difference(P＜0.05).Western Blot results:Compared with the model group,the expression levels of NLRP3,Caspase-1p10 and ASIC3 protein in the in-hibitor group and Zhiqiao Gancao decoction treatment groups were significantly decreased,with statistical difference(P＜0.05).Conclusion:The activation of NLRP3 inflammatory corpuscles can regulate the transcription and function of ASIC3,and thereby construct and maintain the pain sensitivity state of LDH.The mechanismof Zhiqiao Gancao decoction in improving LDH pain is related to its regulation of NLRP3 inflammatory body/ASIC3 signal pathway.
Zhiqiao Gancao decoctionlumbar disc herniationNLRP3 inflammatory bodyacid-sensing ion channels(ASICs)
NETL
NSTL
万方数据
维普
