Intra-Articular Injection of Fructus Ligustri Lucidi Extract Attenuates Pain Behavior and Cartilage Degeneration in Mono-Iodoacetate Induced Osteoarthritic Rats

Abstract

Fructus Ligustri Lucidi (FLL) has been widely used as a traditional Chinese medicine (TCM) for treating soreness and weakness of waist and knees. It has potential for treating OA owing to its kidney-tonifying activity with bone-strengthening effects, but there is so far no report of its anti-OA effect. This study established a rat OA model by intra-articular (IA) injection of mono-iodoacetate (1.5 mg) and weekly treated by IA administration of FLL at 100 μg/mL for 4 weeks. Thermal withdrawal latency, mechanical withdrawal threshold, and spontaneous activity were tested for evaluation of pain behavior, and histopathological (HE, SO, and ABH staining) and immunohistochemical (Col2, Col10, and MMP13) analyses were conducted for observation of cartilage degradation. In vitro effect of FLL on chondrocytes was evaluated by MTT assay and qPCR analysis. Moreover, HPLC analysis was performed to determine its chemoprofile. The pain behavioral data showed that FLL attenuated joint pain hypersensitivity by increasing thresholds of mechanical allodynia and thermal hyperalgesia as well as spontaneous activity. The histopathological result showed that FLL reversed OA cartilage degradation by protecting chondrocytes and extracellular matrix in cartilage, and the immunohistochemical analysis revealed its molecular actions on protein expressions of MMP13, Col2, and Col10 in cartilage. The MTT assay showed its proliferative effects on chondrocytes, and qPCR assay clarified its mechanism associated with gene expressions of Mmp13, Col2, Col10, Adamts5, Aggrecan, and Runx2 in TNF-α treated chondrocytes. Our results revealed an anti-OA effect of FLL on pain behavior and cartilage degradation in OA rats and clarified a molecular mechanism in association with the suppression of chondrocyte hypertrophy and catabolism. IA FLL can be regarded as novel and promising option for OA therapy.

Nanofat lysate ameliorates pain and cartilage degradation of osteoarthritis through activation of TGF-β–Smad2/3 signaling of chondrocytes

“…The pain behavior parameters (PWL and TWL) were measured by a plantar test apparatus (Ugo Basile, Varese, Italy) with the von Frey method ( Chaplan et al, 1994 ; Yan et al, 2018 ). Specifically, daily in the morning, each rat was separately placed in a 10 3 cm individual plastic chamber for 30 min acclimatization, followed by a needling stimulation at the plantar surface of the hind paws, thrice at an interval of 5 min.…”

Investigating the protective effect of tanshinone IIA against chondrocyte dedifferentiation: a combined molecular biology and network pharmacology approach

“…As an early marker of chondrogenesis, Col II contributes to 90% of total collagen content and forms the fibrils, which give cartilage its tensile strength. Furthermore, Col II can protect chondrocytes and the ECM in cartilage by reversing cartilage degradation (39). Chondrocyte hypertrophy represents a late and critical stage of cartilage development.…”

Chondroprotective effects of platelet lysate towards monoiodoacetate-induced arthritis by suppression of TNF-α-induced activation of NF-ĸB pathway in chondrocytes

“…The MWT, TWL, and spontaneous activity were measured as previously described [60]. Briefly, each rat was placed in a wire-mesh based cage for 30 min-acclimatization, followed by needling at its plantar surface of hind paws for three times.…”

“…The cell viability of chondrocytes was evaluated by MTT assay, as previously described [60]. Briefly, the chondrocytes were seeded on 96-well plates for 24 h and treated with PLs derived from 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , and 10 8 platelets for another 24 h and 48 h. MTT solution was added to each well and incubated for 4 h. Subsequently, DMSO was added into each well to dissolve the formazan crystals.…”

“…Then, the reverse transcription was conducted to produce cDNA. As previously applied, the final qPCR reaction system was 20 μl, comprising 10 μl SYBR ® Premix Ex Taq II (Tli RnaseH Plus), 0.4 μl PCR Forward Primer, 0.4 μl PCR Reverse Primer, 1 μl template cDNA and 8.2 μl ddH 2 O, and the qPCR reaction conditions were as follows: 95°C for 5 min for initial denaturation, followed by 40 cycles of denaturation at 95°C for 10 sec, annealing and extension at 60°C for 30 sec [60]. The qPCR assay was performed on an ABI QuantStudio TM 7 Flex Real-Time PCR System (Applied Biosystems; Thermo Scientific, USA).…”