| Objective:Spinal cord injury(SCI)is a serious motor disability disease in the field of spinal surgery and neuroscience,and there is no effective treatment.Traditional Chinese medicine has the advantages of multi-target treatment.It has a definite therapeutic effect on SCI,and has its special advantages.Warming kidney and unblocking Du Channel prescription-" Jisuikang",is based on the,"Kidney-Du deficiency and blood stasis in Du,and failure of pivotal control"theory in pathogenesis of SCI,and screens out from the clinical experience prescription with the effect of"removing blood stasis and removing governor,warming kidney and dampness,and harmonizing Qi and blood".Previous studies have shown that " Jisuikang " can promote the repair of SCI.With the deepening of research,We found it is necessary to explore the regulation mechanism of"Jisuikang " in the treatment of SCI,so as to fully clarify the scientific nature of TCM theory in the treatment of SCI,and explore the therapeutic strategies of TCM in promoting the recovery of nervous function.METHODS:The composition of Chinese medicine compound"Jisuikang"is complex and its clinical use is Chinese medicine water decoction.In order to analyze the potential relationship and interaction between the components of Chinese medicine,and to simplify the prescription scientifically,optimize the water extraction method,and improve the repeatability of the follow-up experiments,We established and quantitatively analyzed the chemical profiles of the water extract of this prescription by high performance liquid chromatography.At the same time,the target screening platform of double luciferase reporter gene system was used to screen the effective components of"Jisuikang"for IRE1/XBP1 pathway,which laid a foundation for the follow-up study of anti-SCI mechanism of"Jisuikang".In order to further clarify the protective mechanism of "Jisuikang" and its effective components against endoplasmic reticulum stress,flow cytometry.RESULTS:The quality control components of the Chinese medicine compound "Jisuikang"were preliminarily established by HPLC.Sixteen components were detected,including ferulic acid,gallic acid,icariin,emodin,chrysophanol,aloe-emodin,plantain,salvianolic acid A,protocatechuic aldehyde,echinacetin,magnoline,quercetin,rhein,salvianolic acid B,hesperidin and paeoniflorin.The target screening platform of double luciferase reporter gene system was used to screen the effective components of"Jisuikang"for IRE1/XBP1 pathway.The results showed that salvianolic acid B,echinarin,plantagin,hesperidin,icariin,paeoniflorin and aloe emodin could effectively promote the splicing activation of XBP1 and specifically deal with endoplasmic reticulum stress through IRE1/XBP1 pathway.The results of CCK8,SRB and flow cytometry showed that thapsigargin could stimulate endoplasmic reticulum stress to induce apoptosis of neuron-like PC 12 cells induced by NGF,and the apoptotic rate was higher.The effective decomposition group,Jisuikang group and icariin group could alleviate endoplasmic reticulum stress induced by carotene and reduce apoptosis of neuron-like PC 12 cells.Western Bloting and q-PCR results showed that the thapsigargin,as an ESR agonist,could indeed up-regulate the expression of XBP1 and IRE 1 in endoplasmic reticulum chaperone molecule Grp78 and ESR signaling pathway,and down-regulate the expression of Grp78,and XBP1,IRE1 protein and mRNA were increased after intervention by Jisuikang,effective decomposition and icariin.Electron transmission microscopy showed that the endoplasmic reticulum of neuron-like PC 12 cells was chaotic and apoptotic after carotene-induced stimulation.The apoptosis of neuron-like PC 12 cells induced by endoplasmic reticulum stress could be ameliorated by effective decomposition group,Jisuikang group and icariin group.By observing the effect of microglia on the phagocytosis of neuronal debris,it was found that the effective decomposition group and Jisuikang group also promoted the phagocytosis of microglia in varying degrees.In terms of the percentage of phagocytosis,there were significant differences between the effective decomposition group and the Jisuikang group compared with the blank group,and there was no significant difference between the LPS group and the effective decomposition group.In terms of the phagocytosis index,there were significant differences between the icariin group,the effective decomposition group,the Jisuikang group and the blank group.CONCLUSION:The effective mechanism of Warming kidney and unblocking Du Channel prescription-" Jisuikang"in the treatment of SCI may be to protect damaged neurons by specifically promoting the splicing activation of XBP1 in ERS.In addition,"Jisuikang"and effective decomposition components can promote microglia to play a scavenger-like role,to swallow neuronal axon fragments,thereby promoting nerve repair function. |
