三色散凝胶贴膏剂的基质处方优化及其对KOA大鼠滑膜炎症的干预效应观察

Optimization of the Matrix Formulation of Sanse Powder Gel Paste and Its Intervention Effect on Synovitis in KOA Rats

目的优化三色散凝胶贴膏剂的基质处方工艺,研究其对膝骨关节炎(KOA)大鼠滑膜炎症的干预效应。方法以综合感官、初黏力、持黏力、剥离强度为评价指标,以聚丙烯酸钠、甘羟铝、酒石酸、甘油为考察因素,采用正交实验筛选最优基质处方。随后,将大鼠随机分为空白组、模型组、三色散凝胶贴膏剂组、三色散传统制剂组、三色散凝胶贴膏剂(含促渗剂)组。采用改良的Hulth法构建KOA模型,造模成功后,各给药组外敷膏药28 d。末次给药后,提取各组大鼠滑膜组织。HE染色评估滑膜炎症,天狼星红染色评估滑膜胶原沉积情况,Krenn评分评价滑膜炎症情况。Western blot和qPCR检测滑膜IL-1β、TNF-α、IL-6蛋白和mRNA表达。结果经过筛选得到三色散凝胶贴膏剂的最优基质处方为聚丙烯酸钠50 g,甘羟铝2.5 g,酒石酸3 g,甘油500 g,三色散150 g,纯水50 mL,卡波姆120 g,薄荷脑36 g。大鼠给药28 d后,HE及天狼星红染色结果显示,与模型组比较,三色散凝胶贴膏剂组和三色散传统制剂组炎性细胞浸润、胶原沉积减少,纤维化程度减轻,Krenn评分显著降低(P<0.01),炎症因子IL-1β、TNF-α、IL-6蛋白和mRNA表达降低(P<0.01);与三色散传统制剂组比较,三色散凝胶贴膏剂组Krenn评分,IL-1β、TNF-α和IL-6蛋白表达无显著差异,IL-1β、IL-6 mRNA表达显著降低(P<0.05),TNF-αmRNA表达无显著差异。与三色散凝胶贴膏剂组比较,三色散凝胶贴膏剂(含促渗剂)组Krenn评分显著降低(P<0.05),IL-1β蛋白表达显著下降(P<0.01),TNF-α和IL-6蛋白表达无显著差异,IL-1β、TNF-α、IL-6 mRNA表达显著下降(P<0.05,P<0.01)。结论优化的基质处方工艺稳定可行,操作方法简便,制备得到的三色散凝胶贴膏剂外观及黏附性良好,且能有效减轻KOA大鼠滑膜炎症,效果不亚于三色散传统制剂。加入促渗剂能增强三色散凝胶贴膏剂的抗炎效果,为进一步研究提供参考。

OBJECTIVE In order to optimize the matrix formulation of Sanse Powder gel paste and to study its anti-synovitis effect on KOA rats.METHODS The comprehensive sensory,initial adhesive strength,holding adhesive strength and peel strength were used as evaluation indexes.Sodium polyacrylate,aluminum glycolate,tartaric acid and glycerol were used as investigation factors.The optimal matrix formulation was screened by orthogonal test.Subsequently,the rats were randomly divided into blank group,model group,Sanse Powder gel paste group,Sanse Powder traditional preparation group and Sanse Powder gel paste(containing penetration enhancers).Except blank group,the modified Hulth method was used to establish the KOA model.After the model was established successfully,plaster was applied externally in each administration group for 28 days.Synovial tissues were extracted from each group after the final administration.Synovitis was assessed by HE staining and synovial collagen deposition was assessed by sirius red staining.Protein and mRNA expressions of IL-1β,TNF-αand IL-6 in synovium were detected by Western blot and qPCR.RESULTS The optimal matrix formulation of the Sanse Powder gel paste was sodium polyacrylate(50 g),aluminium glycolate(2.5 g),tartaric acid(3 g),glycerol(500 g),Sanse Powder(150 g),pure water(50 mL)and carbomer(120 g).After 28 days of drug administration,HE and sirius red staining of synovial tissues results showed that,compared with model group,the inflammatory cell infiltration,collagen deposition and synovial fibrosis in the Sanse Powder gel paste group and the Sanse Powder traditional preparation group decreased,the protein and mRNA expressions of IL-1β,TNF-αand IL-6 were reduced in the Sanse Powder gel paste group and the Sanse Powder traditional preparation group(P<0.01).Compared with Sanse Powder traditional preparation group,there was no significant difference in the protein expressions of IL-1β,TNF-αand IL-6 in the Sanse Powder gel paste group,the mRNA expressions of IL-1βand IL-6 were significantly lower in the Sanse Powder gel paste group(P<0.05),while there was no significant difference in TNF-αmRNA expression.Compared with the Sanse Powder gel paste group,the protein expression of IL-1βwas reduced in the Sanse Powder gel paste(containing penetration enhancers)(P<0.01),while there was no significant difference in protein expressions of TNF-αand IL-6.However,compared with the Sanse Powder gel paste group,the Sanse Powder gel paste(containing penetration enhancers)significantly inhibited the mRNA expressions of IL-1β,TNF-αand IL-6(P<0.05,P<0.01).CONCLUSION The optimized matrix formulation is stable and feasible,and the operation method is simple.The Sanse Powder gel paste has good appearance and adhesion,and can effectively reduce the synovitis of KOA rats.The addition of osmotic enhancer can enhance the anti-inflammatory effect of Sanse Powder gel paste,which provide reference for further research.