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. Author manuscript; available in PMC: 2020 Nov 30.
Published in final edited form as: Biochem Pharmacol. 2019 Mar 9;165:79–90. doi: 10.1016/j.bcp.2019.03.014

Fig. 4. The Ghrelin receptor is required for EM to inhibit cartilage matrix loss and synovitis in vivo.

Fig. 4.

(A) Analysis of total glucose consumption, intracellular ATP levels and reactive oxygen species (ROS) levels at day 2 of culture in healthy and OA human articular chondrocytes. (B) Analysis of total glucose consumption, intracellular ATP levels and reactive oxygen species (ROS) levels at day 2 of culture in nHACs after MIA treatment. (C) RT-qPCR analysis of aggrecan (agg), Col-II, Col-X, IL-1β, TNF⍺ and MMP13 on nHACs treated with MIA (0, 0.1 and 1μM) for 2 days. TBP served as a reference gene for normalization. (D) Safranin O staining analysis of WT and GHSR null mouse knee joints. MIA (50μg/joint) was intra-articularly injected to knee joints at day 0, and samples were harvested at day 7. The degree of cartilage matrix loss was examined according to the OARSI scoring system. (E) Safranin O staining analysis of WT and GHSR null mouse knee joints injected with MIA. MIA was intra-articularly injected to knee joints at day 0. EM (PBS as control) was IP-injected at 50μg/g into the mice daily until sample harvest at day 7. The degree of cartilage matrix loss was examined according to the OARSI scoring system. (F) Synovitis analysis on sections from the medial tibial plateau of WT and GHSR null mice knees injected with EM. MIA was intra-articularly injected to knee joints at day 0. EM (PBS as control) was IP-injected into the mice daily until sample harvest at day 7. Arrowheads indicate thickening of the synovial layers and increased cellularity of resident cells. M = Meniscus. T = Tibia. F=Femur. Synovitis was scored according to established synovitis scoring protocols. Scale bar = 200μm. For A, data were reported as mean ± SD and analyzed by an unpaired t-test. For B and C, data were reported as mean ± SD and analyzed by Dunnett’s test. Comparisons were made between MIA treatments over untreated controls for each gene. For D-F, data were reported as a box plot, where the median, as well as the maximum and the minimum data points were indicated. Data from D-F were analyzed by Kruskal-Wallis statistical test followed by Mann-Whitney U test. * p<0.05. n.s: not significant.