Immune Netw.  2014 Feb;14(1):45-53. 10.4110/in.2014.14.1.45.

Ursodeoxycholic Acid Ameliorates Pain Severity and Cartilage Degeneration in Monosodium Iodoacetate-Induced Osteoarthritis in Rats

• Moon SJ
• Jeong JH
• Jhun JY
• Yang EJ
• Min JK
• Choi JY
• Cho ML

• KMID: 1508828
• DOI: http://doi.org/10.4110/in.2014.14.1.45

Abstract

Osteoarthritis (OA) is a degenerative joint disease characterized by a progressive loss of cartilage. And, increased oxidative stress plays a relevant role in the pathogenesis of OA. Ursodeoxycholic acid (UDCA) is a used drug for liver diseases known for its free radical-scavenging property. The objectives of this study were to investigate the in vivo effects of UDCA on pain severity and cartilage degeneration using an experimental OA model and to explore its mode of actions. OA was induced in rats by intra-articular injection of monosodium iodoacetate (MIA) to the knee. Oral administration UDCA was initiated on the day of MIA injection. Limb nociception was assessed by measuring the paw withdrawal latency and threshold. Samples were analyzed macroscopically and histologically. Immunohistochemistry was used to investigate the expression of interleukin-1beta (IL-1beta), IL-6, nitrotyrosine and inducible nitric oxide synthase (iNOS) in knee joints. UDCA showed an antinociceptive property and attenuated cartilage degeneration. OA rats given oral UDCA significantly exhibited a decreased number of osteoclasts in subchondral bone legion compared with the vehicle-treated OA group. UDCA reduced the expression of IL-1beta, IL-6, nitrotyrosine and iNOS in articular cartilage. UDCA treatment significantly attenuated the mRNA expression of matrix metalloproteinase-3 (MMP-3), -13, and ADAMTS5 in IL-1beta-stimulated human OA chondrocytes. These results show the inhibitory effects of UDCA on pain production and cartilage degeneration in experimentally induced OA. The chondroprotective properties of UDCA were achieved by suppressing oxidative damage and inhibiting catabolic factors that are implicated in the pathogenesis of cartilage damage in OA.

Keyword

Ursodeoxycholic acid (UDCA); Monosodium iodoacetate (MIA); Osteoarthritis; Oxidative stress

MeSH Terms

Administration, Oral
Animals
Cartilage*
Cartilage, Articular
Chondrocytes
Extremities
Humans
Immunohistochemistry
Injections, Intra-Articular
Interleukin-1beta
Interleukin-6
Joint Diseases
Knee
Knee Joint
Liver Diseases
Nitric Oxide Synthase Type II
Nociception
Osteoarthritis*
Osteoclasts
Oxidative Stress
Rats*
RNA, Messenger
Ursodeoxycholic Acid*
Interleukin-1beta
Interleukin-6
Nitric Oxide Synthase Type II
RNA, Messenger
Ursodeoxycholic Acid

Figure

• 

  Figure 1 Effects of UDCA on mechanical hyperalgesia in a model of MIA-induced OA rats. Rats were injected with 3 mg of MIA in the right knee. UDCA was administered orally every day for 14 days after MIA injection. Behavioral tests of mechanical hyperalgesia in MIA-injected rats treated with vehicle or UDCA were evaluated using a dynamic plantar esthesiometer on day 14 (n=6 on each day for each group). The data are expressed as mean and error bars per group. PWL and PWT were conducted right before the administration of UDCA. *p<0.05, **p<0.01 and ***p<0.001 compared with the vehicle-treated OA group.

• 

  Figure 2 Histological evaluation of joints after treatment with UDCA in MIA-induced OA rats. Rats were injected with 3 mg of MIA in the right knee. UDCA was administered orally every day for 14 days after MIA injection. (A) The knee joints from the OA rats treated with UDCA or vehicle were stained with HE, Safranin O, and toluidine blue. The joint lesions were graded on a scale of 0~13 using the modified Mankin scoring system, giving a combined score for cartilage structure, cellular abnormalities, and matrix staining, and an overall Mankin score. The data are expressed as mean and error bars for 6 animals per group. (B) The number of osteoclasts was measured in the subchondral bone legion. *p<0.05, **p<0.01 and ***p<0.001 compared with the vehicle-treated OA group.

• 

  Figure 3 Effects of UDCA on the expression of IL-1β and IL-6 in OA joints. Rats were injected with 3 mg of MIA in the right knee. UDCA was administered orally every day for 14 days after MIA injection. Immunohistochemical staining was used to identify the expression of IL-1β and IL-6 in the articular cartilage. The data are expressed as mean and error bars for 6 animals per group (Original magnification 200×). **p<0.01 and ***p<0.001 compared with the vehicle-treated OA group.

• 

  Figure 4 Attenuated nitrotyrosine and iNOS expression in UDCA-treated OA rats. Rats were injected with 3 mg of MIA in the right knee. UDCA was administered orally every day for 14 days after MIA injection. The knees were resected on day 14 after MIA injection. Immunohistochemical staining was used to identify the expression of nitrotyrosine and induced nitric oxide synthase (iNOS). The data are expressed as mean and error bar for 6 animals per group. (Original magnification 200×). ***p<0.001 compared with the vehicle-treated OA group.

• 

  Figure 5 The decreased catabolic activity in human OA chondrocytes by UDCA treatment. Human articular chondrocytes from OA patients were cultured with or without IL-1β in the absence or presence of UDCA for 48 h following 24 h of starvation. The mRNA expression of catabolic factors, MMP-3, MMP-13 and ADAMTS5, was measured by quantitative real-time PCR. β-actin was used as the internal control (A). The cell viability was determined by MTT assay (B). There was no cytotoxic effect in the doses of UDCA (ranging from 0.1 to 50µM) that were studied in this experiment. The data are expressed as dot plots with mean (bar) for three independent experiments per group. *p<0.05, **p<0.01 and ***p<0.001 compared with the untreated IL-1β-stimulated chondrocytes.

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