Optimization of the extraction technology for the raw drugs of Sanse powder gel paste / 中国药房

OBJECTIVE To optimize the extraction technology for the raw drugs of Sanse powder gel paste. METHODS SD rats were divided into blank group， model group， traditional technology group， water extraction group and ethanol extraction group， with 5 rats in each group. Anterior cruciate ligament transection was used to construct knee osteoarthritis model， and the pharmacodynamic effects of different extraction methods on arthritic rats were investigated. Analgesic experiments were conducted using cold and hot pain thresholds and pain mediators calcitonin gene-related peptide （CGRP）， cyclooxygenase-2 （COX-2）， substance P （SP）， and prostaglandin E2 （PGE2） contents as indicators. HE staining was performed on the synovial membrane of rats to observe the degree of synovial cell proliferation， inflammatory infiltration and vascular invasion， and anti-inflammatory experiments were conducted using protein and mRNA expressions of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α） and IL-6 as indicators. The analgesic and anti-inflammatory effects were compared among those groups. In the orthogonal test， ethanol dosage， extraction time and extraction times were used as evaluation factors， and the contents of casticin， strychnine and toxiferine were taken as evaluation indicators； comprehensive score was calculated. The validation experiments were carried out after optimizing the extraction technology of the raw drugs of Sanse powder gel paste. RESULTS Compared with the model group， the cold and heat pain thresholds of drug administration groups （except for the traditional technology group） were all increased significantly （P＜0.05）， while the contents of pain （No.Y2021rc02） mediators CGRP， COX-2， SP and PGE2 were all decreased significantly （P＜0.05）. HE staining showed that inflammatory cell infiltration， fibrosis and collagen deposition were 炎。E-mail：liuzixiu3221@126.com decreased in the administration groups； a small amount of capillary proliferation could be found； the protein and mRNA expressions of inflammatory factors such as IL-1β， IL-6 and TNF-α were decreased significantly in synovial tissue of rats in administration groups （P＜0.05）. Compared with the traditional technology group， most indicators of the ethanol extraction group were significantly reduced （P＜0.05）， and only heat pain threshold and mRNA expression of IL-6 in rats were decreased significantly in the water extraction group （P＜0.05）. The optimal extraction technology of the raw drugs of Sanse powder gel paste included suitable dose of Sanse powder， 8-fold 55% ethanol， heating reflux extraction for 90 minutes， extracting twice. The results of 3 times of verification experiments showed that the average contents of casticin， strychnine and toxiferine were 0.007%， 0.092%， and 0.214%， respectively； RSD were all less than 5%. CONCLUSIONS The optimized extraction technology for the raw drugs of Sanse powder gel paste is stable and feasible， which can improve the efficacy of the preparation.

Discussion on the Mechanism of Synovitis of KOA Relieved by “Sanse Powder ”Volatile Oil Based on the Activation of NLRP 3 Inflammasome / 中国药房

OBJECTIVE：To discuss the mechanism of synovitis of knee osteoarthritis （KOA）relieved by “Sanse powder ” volatile oil based on the activation of Nod-like receptor family 3（NLRP3）inflammasome. METHODS ：“Sanse powder ”volatile oil was extracted by distillation- condensation process and the components of it were analyzed by GC-MS. Fibroblast-like synovial cells（FLS）were extracted from the knee joint of male SD rats. The effects of 10，25，50，100，250，500，1 000 μg/mL“Sanse powder”volatile oil on the viability of FLS were examined by CCK-8 assay. KOA inflammatory cell model was induced by lipopolysaccharide for 12 h. The effects of 10，250 μg/mL“Sanse powder ”volatile oil on the protein and mRNA expression of NLRP3，caspase-1 and apoptosis-associated speck-like protein （ASC）were detected. The levels of IL- 1β and IL-18 in supernatants of FLS were determined . RESULTS ：Totally 0.51-0.61 g volatile oil with yellow clear and unique aromatic odor were extracted from“Sanse powder ”，and the extraction rate was 0.33％-0.41％（n＝9）. A total of 41 chemical components were isolated ，and 30 of them were identified and their peak area accounted for 90.073 6％ of the total peak area. The components with high relative content were gingerol flavonoids （17.573 9％），δ-junionene（15.434 5％），gingerol（11.509 5％），etc. When the concentration of volatile oil was 10-250 μg/mL，there was no significant effect on survival rate of FLS （P＞0.05）. Compared with blank group ，the relative protein and mRNA expression of NLRP 3，caspase-1 and ASC and the levels of IL- 1β and IL-18 in supernatant were significantly increased in model group （P＜0.05）. Compared with model group ，relative expression or levels of above indexes were all decreased significantly in FLS and supernatant of “Sanse powder ”volatile oil 10 and 250 μg/mL groups（P＜0.05）. CONCLUSIONS：“Sanse powder ”volatile oil can inhibit NLRP 3 inflammasome activation in FLS and reduce the downstream inflammatory cascade response ，thus exerting its efficacy in ameliorating synovial inflammation of KOA.