Application of salvianic acidA in preparation of medicine for relieving occurrence and development of lumbar facet joint osteoarthritis
A technology of danshensu and joint bone, applied in the field of biomedicine, can solve the problems of lumbar facet joint osteoarthritis and unclear mechanism
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Embodiment 1
[0025] Effects of Danshensu Sodium (SAAS) on Cartilage and Synovial Tissue of Lumbar Facet Joint Osteoarthritis (LFJOA) Model in Vivo
[0026] (1) In vivo LFJOA model grouping and modeling
[0027] Grouping: 30 marked SD rats were randomly divided into 3 groups, 10 in each group, respectively: control group (Sham group), treatment group (CLG group), experimental group (CLG+SAAS group)
[0028] Modeling: LFJOA rat model was established by induction of type II collagenase. The rats anesthetized by intraperitoneal injection of chloral hydrate were fixed on the operating table and routinely disinfected with povidone iodine. During the operation, a 3cm incision was made in the center of the back, and the right paravertebral muscles were peeled off to expose the three small joints on the right side L3-L4, L4-L5, and L5-L6. 6U type II collagenase was injected into the above three joint cavities of the treatment group and the experimental group respectively with a micro-syringe with...
Embodiment 2
[0041] Regulatory effect of danshensu sodium (SAAS) on the TNFR1 pathway of chondrocyte apoptosis and the inflammatory NF-κB pathway of fibroblast-like synoviocytes
[0042] (1) In vitro cell model establishment
[0043] ①Apoptotic model of chondroma cell SW1353
[0044] Human chondrosarcoma cells SW1353 were cultured in 10% FBS serum + DMEM at 37°C, 5% CO 2 Cultured in an incubator, stimulated with 50ng / ml TNF-α for 12 hours, then detected the expression of apoptosis-related key proteins by flow cytometry, TUNEL staining, Caspase3 / 8 kinase activity experiment and WB, so as to observe the apoptosis of SW1353 cells.
[0045] ②Fibroblast-like synoviocytes (FLSs) inflammation model
[0046] 1) Isolation and culture of fibroblast-like synoviocytes:
[0047] Rats were sacrificed under anesthesia and exposed to the synovial tissue at the lumbar facet joints in an ultra-clean table. The excess fat and connective tissue were removed and placed in a centrifuge tube, and the synovial...
Embodiment 3
[0060] E3 ligase and target screening by in vitro cell experiments to clarify the regulatory effect of danshensu sodium (SAAS) on TRIP and the molecular mechanism of SAAS synergistic effect
[0061] (1) E3 ligase and target screening
[0062] In SW1353 cells and primary FLS cell culture medium, after adding 10 μM SAAS, the mRNA expressions of osteoarthritis and apoptosis-related E3 ligases such as Cbl-b, TRIM22, TRIM25, TRIM38, TRIM41, TRIM56 and TRIP were detected by qPCR Changes. At the same time, in SW1353 cells and FLS cell culture medium, after adding gradient doses of SAAS (2.5, 5, 10 μM), the protein expression of E3 ligase was detected by WB. The E3 ligase specifically regulated and expressed by SAAS was screened out, namely TRIP.
[0063] ① Co-immunoprecipitation (co-IP)
[0064] In the SW1353 cell apoptosis model and the FLSs cell inflammation model, the combination of TRIP and the target molecule (such as RIP1) was detected respectively. The constructed TRIP mol...
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