消瘀散精简方新组方凝胶贴透皮成分的探究及其对骨关节炎大鼠软骨损伤和滑膜炎的治疗作用OA

Objective:To study the transdermal absorption of ingredients,therapeutic effect and biosafety of the new formula gel patch compound of the Xiaoyusan in osteoarthritis(OA)rats.Methods:The skin transdermal absorption process was simulat-ed by Franz diffusion cell method,and the new formula gel patch compound of the Xiaoyusan receiving solution was analyzed by UPLC-Q-TOF/MS.24 rats were divided into sham operation group,OA model group,positive control group(Flurbiprofen Cata-plasms)and experimental group(the new formula gel patch compound of the Xiaoyusan)with 6 rats in each group.All rats under-went anterior cruciate ligment transection(ACLT)except the sham operation group.The OA rat model was established by ACLT.One week after the model was made,the new formula gel patch compound of the Xiaoyusan was applied to the knee joint of the rat as an experimental drug.Orbital blood and knee joint specimens were collected after continuous treatment for 21 days,and hematoxylin-eosin(H&E)staining and safranin O-fast green(Safranin O)staining were used to analyze the severity of knee cartilage injury in each group.H&E staining was used to evaluate the severity of knee synovitis.The expression of collagen type Ⅱalpha 1(Col2a1),matrix metallopeptidase 13(MMP13),and caspase 3 in cartilage,as well as the expression of inducible nitric oxide synthase(iNOS)and tumour necrosis factor alpha(TNF-α)in synovium,was detected using immunohistochemistry in each group;enzyme linked immunosorbent assay(ELISA)was used to analyze the levels of serum inflammatory cytokines TNF-α and interleuin 17(IL-17)in each group.The tissue and organ structure of each group was analyzed by H&E staining to evaluate the biosafety.Results:(1)The new formula gel patch compound of the Xiaoyusan contain 28 prototype ingredients,in-cluding 6-methyl-rhein and rhein in imperial medicine Rhubarb.(2)After 21 days of external application treatment,both the new formula gel patch compound of the Xiaoyusan and Flurbiprofen Cataplasms can save articular cartilage injury and synovial inflam-mation in OA rats,and the therapeutic effect of the new formula gel patch compound of the Xiaoyusan is better.The expression of Col2a1 was significantly up-regulated,and the expressions of MMP13 and Caspase 3 were significantly inhibited(P<0.05).(3)The treatment of flurbiprofen gel paste and the new formula gel patch compound of the Xiaoyusan can alleviate synovial inflam-mation in rats.Immunohistochemical staining further indicated that the expression of TNF-α and iNOS in synovial membrane of rats in the new formula gel patch compound of the Xiaoyusan group was significantly down-regulated(P<0.05).(4)ELISA re-sults showed that the occurrence and development of OA and the treatment of the new formula gel patch compound of the Xiaoyu-san did not cause the up-regulation of systemic inflammatory factors TNF-α and IL-17.Meanwhile,after external treatment of the new formula gel patch compound of the Xiaoyusan,the microstructure of tissues and organs of rats in each group were normal and there was no obvious inflammatory infiltration.Conclusion:The new formula gel patch compound of the Xiaoyusan contains 6-methyl-rhein and rhein,which are found in rhubarb.The new formula gel patch compound of the Xiaoyusan can promote the syn-thesis of cartilage matrix,inhibit the apoptosis of chondrocytes and the expression of synovial inflammatory factors in the treatment of OA,and has good biosafety.It is expected to further promote clinical conversion through safety and efficacy evaluation.