• Herbal Formula Science
• /
• v.26 no.4
• /
• pp.283-294
• /
• 2018

Objectives : Mahaengeuigam-Tang (MHEGT) has been used as a traditional medicine for the treatment of rheumatic aerthritis, rheumatisim, eczema and asthma. The aim of this study was to investigate the molecular mechanisms of MHEGT for cartilage protection in monosodium iodoacetate(MIA)-induced osteoarthritis, particularly focusing on apoptosis. Method : Thirty young male Sprague-Dawley rats were used for the study. Rats were intra-articularly injected with 2 mg MIA in a total volume of 50 ㎕ saline. In MHEGT group, MHEGT extract was orally administered once daily to MIA-induced osteoarthritis rats, and rats of control group were given with saline only. At 4 weeks after MIA injection, all animals were sacrificed, and the histological changes and articular thickness were assessed by hematoxylin and eosin staining. Moreover, the immunohistochemical analyses of BAX and Bcl-2 were carried out. Results : The histomorphological examinations revealed that MHEGT reduced MIA-induced cartilage damage. And, MHEGT ameliorated the severity of cartilage surface damages after MIA injection. Furthermore, MHEGT suppressed the MIA-induced increases of pro-apoptotic BAX protein and increased the protein expression of anti-apoptotic Bcl-2 protein. Conclusion : These findings indicate that MHEGT protects against MIA-induced cartilage damage by inhibition of the apoptotic pathway, demonstrating significant protection of cartilage against osteoarthritis. These results suggest that MHEGT may potentially have clinical applications in the treatment of osteoarthritis.

• Journal of Life Science
• /
• v.33 no.6
• /
• pp.455-462
• /
• 2023

During pregnancy, maternal immune activation (MIA) from infection increases the risk of neurodevelopmental diseases, including schizophrenia and autism spectrum disorders. MIA induced by polyinosinic-polycytidylic acid (poly (I:C)) and lipopolysaccharide (LPS) in animal experiments has led to offspring with abnormal behaviors and brain development. In addition, it has recently been reported that microglia, which reside in the brain and function as immune cells, play an important role in behavioral abnormalities and brain development in MIA-induced offspring. However, the underlying mechanism remains unclear. In this study, we investigated whether microglia-specific inhibition of GPR56, a member of the G protein-coupled receptor (GPCR) family, causes behavioral abnormalities in brain development. First, MIA induction did not affect the microglia population, but when examining the expression of microglial GRP56 in MIA-induced fetuses, GPR56 expression was inhibited between embryonic days 14.5 (E14.5) and E18.5 regardless of sex. Furthermore, microglial GPR56-suppressed mice showed abnormal behaviors in the MIA-induced offspring, including sociability deficits, repetitive behavioral patterns, and increased anxiety levels. Although abnormal cortical development such as that in the MIA-induced offspring were not observed in the microglial GPR56-suppressed mice, their brain activity was observed through c-fos staining. These results suggest that microglia-specific GPR56 deficiency may cause abnormal behaviors and could be used as a biomarker for the diagnosis and/or as a therapeutic target of behavioral deficits in MIA offspring.

• Journal of Korean Medicine Rehabilitation
• /
• v.26 no.3
• /
• pp.1-15
• /
• 2016

Objectives This study was designed to evaluate the anti-inflammatory effects of Seungseup-tang (SST) on the monosodium iodoacetate (MIA)-induced osteoarthritis rats. Methods Osteoarthritis was induced by injection of MIA ($50{\mu}l$ with 80 mg/ml) into knee joint cavity of rats. Rats were divided into 4 groups (normal group, control group, indomethacin treated group, SST treated group, each n=6). Normal group was not injected with MIA and taken normal diet. Control group was injected with MIA and taken with distilled water. Indomethacin treated group was injected with MIA and taken indomethacin 5 mg/kg by oral administration. SST treated group was injected with MIA and taken SST 200 mg/kg by oral administration. We examined the weight-bearing ability of hind paw, biomarkers related to oxidative stress in serum, inflammatory proteins and inflammatory mediators and cytokines. Moreover, histopathological examination of knee joint structure was also performed by Hematoxylin & Eosin (H&E), Safranin-O staining method. Results In the present study, SST treated group showed a similar inhibitory effects alike indomethacin treated group, in most of the studied parameters of inflammation. The increased oxidative stress biomarker such as reactive oxygen species (ROS) and peroxy nitrite ($ONOO^-$) in the serum were reduced with SST. Especially, the level of $ONOO^-$ compared with control group significantly suppressed. Also, the expression of inflammatory mediators and cytokines induced by nuclear factor-kappa B (NF-${\kappa}B$) activation was modulated through inhibition of IkBa phosphorlation. In addition, histological analysis revealed that cartilage damage by MIA repaired markedly in SST treated group. Conclusions According to the results, Seungseup-tang may be effective for preventing the progression of osteoarthritis.

• Journal of Korean Medicine Rehabilitation
• /
• v.25 no.2
• /
• pp.51-64
• /
• 2015

Objectives This study was carried out to investigate the effects of Gyejigabuja-tang extracts on the Monosodium iodoacetate (MIA) induced osteoarthritis in rats. Methods Osteoarthritis was induced by injection of MIA into knee joint cavity of rats. Rats are divided into 4 groups (normal, control, positive comparison group, GBT group, each n=5). Normal group was injected by normal saline into knee joint cavity only. Control group was induced for osteoarthritis by MIA and oral medicated with distilled water. Positive comparison group was injected with MIA and taken Joins tablet 25 mg/kg. GBT group was injected with MIA and taken Gyejigabuja-tang extracts 300 mg/kg. Positive comparison group and GBT group were oral medicated for each substance once a day for 4 weeks. ALT, AST and creatinine were evaluated for hepatotoxicity and nephrotoxicity. Hind paw weight bearing ability was examined and inflammatory cytokines (IL-$1{\beta}$, IL-6, TNF-${\alpha}$), $PGE_2$, $LTB_4$, osteocalcin and deoxypyridinoline (DPD) within serum were analysed. Knee joint structures were observed by Hematoxylin & Eosin (H&E), Safranin-O staining method. Results 1. Function of liver and kidney was not affected. 2. Hind paw weight bearing ability was significantly improved. 3. IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ in experimental group were significantly decreased compared with control group. 4. $PGE_2$, $LTB_4$, Osteocalcin and DPD in experimental group were decreased compared with control group. 5. In histopathologic observation, injury on synovial membrane and cartilage of experimental group was lesser than control group (H&E, Safranin-O staining). Conclusions Based on these results, it can be suggested that Gyejigabuja-tang has anti-inflammation effects on the MIA-induced osteoarthritis in rats.

• Journal of Korean Medicine Rehabilitation
• /
• v.25 no.2
• /
• pp.37-50
• /
• 2015

Objectives The purpose of this study was to know the effects of Danggwisayeok-tang (Dangguisinitang) extract (DGSYT) on monosodium iodoacetate (MIA)-induced rat osteoarthritis. Methods For this purpose, rats were divided into 5 groups. Normal group was not injected with MIA and orally administered any medication. Control group was injected with MIA and not orally administered any medication. DGSYT100 group was injected with MIA and orally administered 100 mg/kg of DGSYT. DGSYT300 group was injected with MIA and orally administered 300 mg/kg of DGSYT. JoinsT group was injected with MIA and orally administered 20 mg/kg of Joins tablet. DGSYT100 and DGSYT300 groups were orally administered DGSYT during a week before and 3 weeks after based on the day MIA injected. The changes of hepatotoxicity, nephrotoxicity, relative hind paw weight distribution, cytokine in serum, cytokine messenger ribonucleic acid (mRNA) in joint tissue and histopathological observation (Hematoxylin & Eosin and Safranin-O staining) were measured. Results Alanine aminotransferase (ALT) levels of DGSYT100, DGSYT300 and JoinsT groups were increased significantly, but these results were within normal range. Aspartate aminotransferase (AST) and creatinine levels of all groups were not changed significantly. In the change of relative hind paw weight distribution, DGSYT300 and JoinsT groups were decreased significantly 14 and 21 days after MIA injected. Interleukin-$1{\beta}$ (IL-$1{\beta}$) and Interleukin-6 (IL-6), Leukotriene $B_4$ and Osteocalcin levels of DGSYT300 and JoinsT groups were decreased significantly. In measurement of IL-$1{\beta}$ and nitric oxide synthase-II mRNA relative quantitative of control, DGSYT100, DGSYT300 and JoinsT groups were decreased significantly. In measurement of TNF-${\alpha}$, IL-6 and Cyclooxygenase-2 mRNA relative quantitative of control, DGSYT300 and JoinsT groups was decreased significantly. In histopathological observation of knee, synovial tissue, cartilage and proteoglycan of DGSYT100, DGSYT300 and JoinsT were well preserved compared with control group. Conclusions According to the results, DGSYT has anti-inflammation and pain relief effects. So it should be suppressed progression of arthritis in MIA-induced osteoarthritis rat.

• The Journal of Korean Medicine
• /
• v.34 no.3
• /
• pp.69-85
• /
• 2013

Objectives: This study investigated the anti-osteoarthritic effects of Kyejiinsam-tang (hereinafter referred to KIT) on the monosodium iodoacetate (MIA)-induced osteoarthritis rats. Methods: Anti-oxidative effects of KIT were measured by scavenging activities of DPPH, reactive oxygen species (ROS) and nitric oxide (NO). Scavenging activities of anti-oxidation in lipopolysaccharide (LPS)-treated RAW 264.7 cells were also measured for inhibitory effects against the production of inflammatory mediators (tumor necrosis factor-${\alpha}$, interleukin-$1{\beta}$, interleukin-6). Osteoarthritis was induced in rats by injecting MIA in the knee joint. Rats were divided into a total of 4 groups (n=6). The normal group were not treated at all without inducing osteoarthritis whereas the control group were induced for osteoarthritis by MIA and oral medicated physiological saline per day. The positive comparison group was injected with MIA and after 7 days, 2 mg/kg of Indomethacin. The experimental group was injected with MIA and after 7 days was medicated with 34 mg/kg of KIT. Indomethacin and KIT were orally-medicated for each substance a total of 4 weeks, once per day. Weight-bearing on hind legs was measured every week after MIA injection. At the end of the experiment (5 weeks after MIA injection), micro CT (computed tomography)-arthrography and histopathological examinations on the articular structures of knee joint were performed. The effect on inflammatory cytokines and immunological cells in synovial fluid was measured. Volume of cartilage was measured by micro CT-arthrography. Injury to synovial tissue was measured by H & E (hematoxylin and eosin), Safranin-O immunofluorescence. Results: 1. Cytotoxicity against hFCs was insignificant. 2. KIT showed the potent full term for DPPH. 1. NO was significantly reduced by KIT (at 100, $200{\mu}g/m{\ell}$) and ROS was also reduced, but not significantly, by KIT (at $200{\mu}g/m{\ell}$). 2. IL-6 and IL-$1{\beta}$ were significantly reduced by KIT (at 100, $200{\mu}g/m{\ell}$) and TNF-${\alpha}$ was also reduced, but not significantly, by KIT (at $200{\mu}g/m{\ell}$). 1. In hind legs weight-bearing measurement, level of weight increased. 2. Functions of liver and kidney were not affected. 3. IL-$1{\beta}$ was significantly reduced and TNF-${\alpha}$, IL-6 were also reduced but not significantly. 4. PGE2 (prostaglandin E2), LTB4 (leukotriene B4) were significantly reduced in the KIT group. 5. MMP-9 (matrix metalloproteinase-9), TIMP-1 (tissue inhibitor of metalloproteinases-1) and Osteocalcin were significantly reduced in the KIT group. 6. Destruction of cartilage on micro CT arthrography was reduced but had no significant differences. 7. Histopathologically, injury to synovial membrane of the KIT group was decreased and proteoglycan content of KIT group was increased. Conclusions: According to this study, Kyejiinsam-tang has inhibiting effect on the progression of arthritis in MIA-induced osteoarthritis rat. Kyejiinsam-tang has anti-oxidants and anti-inflammation effects, and is related to inhibiting the activity of inflammatory cytokine and injury of volume in cartilage.

• Journal of the Korean Society of Physical Medicine
• /
• v.9 no.2
• /
• pp.151-159
• /
• 2014

PURPOSE: The purpose of this study was to evaluate whether light-emitting diodes (LED) irradiation could be effective in a noninvasive, therapeutic device for the treatment of osteoarthritis(OA). METHODS: Twenty-four male Sprague-Dawley rats were divided into four groups: Vehicle control (saline); monosodium iodoacetate-injection (MIA); LED irradiation after MIA injection (MIA-LED); indomethacin-treatment after MIA injection (MIA-IMT). OA was induced by intra-articular injection of 3 mg MIA through the patellar ligament of the right knee. Vehicle control rats were injected with an equivalent volume of saline. The LED was irradiated for 15 min/day for a week after 7 days of MIA treatment. To compare with the effect of LED irradiation, the indomethacin was administrated 20 mg/kg twice a week orally after 7 days of MIA treatment. Knee joints were removed and fixed overnight in 10% neutral buffered formalin and decalcified by EDTA for 2 week before being embedded in paraffin. The assessment of OA induction were monitored by knee movement and radiographic finding. Histologic analysis were performed following staining with hematoxylin and eosin, safranin O-fast green, or toluidine blue, picrosirius red, and histologic changes were scored according to a modified Mankin system. Apoptotic cell in tissue sections was detected using TUNEL method. RESULTS: Radiographic examination could not show the differences between the MIA-treated and the MIA-LED-treated rats. In the histologic analysis, however, LED irradiation prevented cartilage damage and subchondral bone destruction, and significantly reduced mononuclear inflammatory cell infiltration and pannus formation. LED irradiation also reduced apoptosis of cartilage cells, but it prevented apoptosis of infiltrated inflammatory cells in synovium. In addition, LED irradiation showed an increase of collagen production in the meniscus. CONCLUSION: These results suggest that the 840 nm LED irradiation would be a suitable non-thermal phototherapy for the treatment of OA, as a cartilage protection and anti-inflammatory modality.

• Journal of Korean Medicine Rehabilitation
• /
• v.24 no.1
• /
• pp.31-45
• /
• 2014

Objectives This study intends to clarify how Leejung-tang (here in after reffered to LJT) affect Wistar Rat whose osteoarthritis was induced by MIA. Methods Osteoarthritis was induced into rat by injecting MIA in its knee joint. Rats are divided into a total of 4 groups (n=6). Normal group are not treated at all without inducing osteoarthritis whereas control group were induced for osteoarthritis by MIA and oral medicated with 2 ml of physiological saline per day. Positive comparison group (Indomethacin) was injected with MIA and after 7 days, 2 mg/kg of Indomethacin was medicated. Experimental group (LJT) was injected with MIA and after 7 days that was medicated with 23 mg/kg of LJT. Indomethacin and LJT were oral medicated for each substance a total of 4 weeks with one time per day. After experiments (from 1 week after injection of MIA to 4 weeks elapsed), Hind paw weight bearing ability, Functions of liver and kidney, Serum prostaglandin $E_2$, TNF-${\alpha}$, IL-1${\beta}$, IL-6, Osteocalcin, TIMP-1, MMP-9, LTB4 and amount of cartilage were measured and histopathological variations for knee joint structures were observed. Results 1) Hind paw weight bearing ability of LJT administration group was increased but there was no statistical significance. 2) Functions of liver and kidney were not affected. 3) Serum prostaglandin $E_2$, IL-1${\beta}$, Osteocalcin, MMP-9 were significantly decreased and TNF-$\alpha$, IL-6, TIMP-1, LTB4 were also decreased but there were no statistical significance. 4) In H&E staining and Safranin-O staining, there were small histopathological changes in LJT administration group than control group. 5) In micro CT (computed tomography)-arthrography, cartilage destruction was more suppressed in LJT administration group than control group. Conclusions Based on all results mentioned above, Leejung-tang (LJT) is believed to be meaningful for suppressing the progress of osteoarthritis and its treatments because of its anti-inflammatory effects and alleviation of pain with histopathological effective efficacy.

• Journal of Acupuncture Research
• /
• v.33 no.2
• /
• pp.35-49
• /
• 2016

Objectives : This study was designed to investigate the anti-osteoarthritic effects of clematidis radix pharmacopuncture on the MIA-induced osteoarthritis rats. Methods : The subjects were divided into 4 groups ; Normal rat group(N, n=5), MIA-induced osteoarthritis control rat group(C, n=5), MIA-induced osteoarthritis rat group injected with normal saline at ST36(NS, n=5), and MIA-induced osteoarthritis rat group injected with clematidis radix pharmacopuncture at ST36(CR, n=5). The experiment was conducted over a period of 21 days after injecting MIA. We analyzed body weight, hind paw weight distribution, liver and renal function, immunocytes, cykokines, inflammatory mediators, inflammatory proteins and mRNA expressions, as well as histopathological changes. Results : The CR group showed a significant increase in the hind paw weight distribution, and significant decreases in IL-$1{\beta}$, IL-6, $PGE_2$, $LTB_4$, osteocalcin, deoxypyridinoline level, the protein expression of COX2, arachidonate 5 lipoxygenase, and the mRNA expression of COX2, TNF-${\alpha}$, IL-$1{\beta}$, and NOS-II. In terms of the joint damages induced by osteoarthritis, the CR group showed a greater protective effect than group C in histopathologic observation (H&E, Safranin-O staining). Conclusion : These results demonstrated that clematidis radix pharmacopuncture had anti-inflammatory and analgesic effects. In addition, these results showed that it inhibited the progression of osteoarthritis.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.27 no.4
• /
• pp.158-176
• /
• 2014

Purpose : The purpose of this study is to investigate the effect of ARTEMISIAE CAPILLARIS HERBA and COPTIDIS RHIZOMA Extracts on cell death in pancreatic cancer cells. Method : Human-derived pancreatic cancer cell line, MIA PaCa-2 cells were treated by Prescription A with various concentrations and the cytotoxicity was determined by MTT assay. To investigate the effects of Prescription A on pancreatic cancer cells, the staining of Annexin V/PI, cell cycle arrest, nuclear chromatin condensation and the production of reactive oxygen species (ROS) were examined. The effect of Prescription A's effective components, ARTEMISIAE CAPILLARIS HERBA and COPTIDIS RHIZOMA Extracts on cell death were also observed. Results : The viability of MIA PaCa-2 cells treated with Prescription A were decreased in a dose dependent manner. Prescription A induced cell death in MIA PaCa-2 cells as shown by result of Annexin V/PI double staining, chromatin condensation and cell cycle arrest. In addition, production of ROS was increased by Prescription A treatment, suggesting that ROS induced by Prescription A mediated cell death. Furthermore, Prescription A's effective components, ARTEMISIAE CAPILLARIS HERBA and COPTIDIS RHIZOMA Extracts were also induced apoptosis of MIA PaCa-2 cells through ROS production. Conclusion : These results suggest that Prescription A's effective components, ARTEMISIAE CAPILLARIS HERBA and COPTIDIS RHIZOMA Extracts induced death of MIA PaCa-2 through ROS production.