Labisia pumila prevented osteoarthritis cartilage degeneration by attenuating joint inflammation and collagen breakdown in postmenopausal rat model

Abstract

The tropical herb Labisia pumila is traditionally used in facilitating childbirth and post-partum care. The effects of L. pumila leaf extract (LP) in explant cartilage culture and on postmenopausal osteoarthritis (OA) rat model were assessed. The LP (10, 25 and 50 µg/ml) or diclofenac (10 µg/ml) was added to the cartilage explants containing bovine IL-1β (20 ng/ml), to evaluate their direct effects on cartilage degradation. In the preclinical study, rats were grouped (n = 8) into: non-treated OA; OA + diclofenac (5 mg/kg); OA + LP extract (150 and 300 mg/kg); and healthy control. To induce OA, monosodium iodoacetate was injected into the ovariectomised female rats' intra-articular knee joints and evaluated for OA severity after 8 weeks via physical (radiological, macroscopic and histological observations), biochemical, ELISA and mRNA expression analysis (for inflammation and cartilage degradation biomarkers). The LP reduced the nitric oxide and proteoglycan release from the cartilage explants under IL-1β induction. The radiological, macroscopic, microscopic and histological images showed the OA rats treated with LP and diclofenac had significantly reduced osteophytes and cartilage erosions compared to non-treated OA rats. The extract significantly up-regulated the anti-inflammatory interleukin-10, collagen type II and down-regulated pro-inflammatory PTGS2 (prostaglandin-endoperoxide synthase 2) mRNA expressions compared to non-treated control. The LP treatment significantly reduced serum collagenases (MMP-1 and MMP-3) and collagen type II degradation biomarker (CTX-II) levels in OA rats. The LP containing myricetin and gallic acid suppressed inflammation, collagenases and cartilage degradation, and helped cartilage matrix synthesis, to prevent OA at the dose equivalent to 30-60 mg/kg daily for humans.

Changes of type II collagenase biomarkers on IL‑1β‑induced rat articular chondrocytes

“…MMP-13 is a collagenase that regulates matrix degradation and binds to type II collagen in the ECM (7,38). In addition, proteolytic enzymes and MMPs could be stimulated by IL-1β, which are important factors for cartilage destruction (13). The results of a previous study on type II collagen degradation in chondrocytes were consistent with those of the present study (4).…”

“…When Col II breaks down and produces fragments, such as collagenase, type II collagen cleavage neoepitope (C2C) and collagen type-II C-telopeptide fragments (CTX-II) are generated (11,12). C2C is a useful biomarker that reflects the cleavage of Col II in OA and the levels of CTX-II have been demonstrated to be associated with the total damage of osteophytes (13).…”

Epicatechin and scopoletin richMorinda citrifolia(Noni) leaf extract supplementation, mitigated Osteoarthritis via anti-inflammatory, anti-oxidative, and anti-protease pathways

“…The ex‐vivo explant culture method was adapted to demonstrate if the extract, scopoletin, Diclofenac, or epicatechin could directly inhibit cartilage degradation in the presence of the inflammatory IL‐1β (Madzuki, Lau, Che Ahmad Tantowi, Mohd Ishak, & Mohamed, ). The fresh bovine cartilage disks ( n = 6 disc cartilages per well, triplicate wells per group) were grouped into (a) normal control (N), (b) non‐treated inflammation control (NT), (c) diclofenac drug 20 μg/well (DRUG), (d) M. citrifolia at 1, and 2.5 μg/well (MC1; MC2.5), (e) scopoletin at 1 and 5 μg/well (SC2; SC5) (f) epicatechin at 1 and 5 μg/well (EP1; EP5); and further incubated for 12 hr.…”

Vernonia amygdalina inhibited osteoarthritis development by anti‐inflammatory and anticollagenase pathways in cartilage explant and osteoarthritis‐induced rat model

“…After 96 hr, the explants were washed in PBS (3 × 3 min) and replaced with fresh DMEM/F12/4% penicillin/streptomycin mixture, and interleukin 1β (IL‐1β, 20 ng/ml) was added to aggravate stress/inflammation. The explants were treated with diclofenac (10 μg/ml; Mepharm Sdn Bhd, Subang Jaya, Malaysia) or VA leaf extract (0, 10, 25, or 50 μg/ml) and cultured for 120 hr under 5% CO 2 at 37°C; then the media were collected and stored at −20°C for proteoglycan and nitric oxide release analyses (Madzuki, Lau, Che Ahmad Tantowi, Mohd Ishak, & Mohamed, ). The proteoglycan release was determined using 1‐9‐dimethyl‐methylene blue proteoglycan detection kit from Astarte Biologics (Washington, USA).…”

“…Thermal cycling and fluorescence detection were performed using a CFX96 Touch qPCR System (Bio‐Rad, USA). The relative mRNA levels were determined using 2 −ΔCt method (the difference between cycle threshold value for gene of interest and a control housekeeping gene that do not change much with the treatments, calculated in logarithm base 2), which were normalized to the housekeeping gene glyceraldehyde‐3‐phosphate dehydrogenase (NM_017008; Madzuki et al, ).…”