Objectives : This study was built to investigate the effect of Trachelospermi Caulis herbal-acupuncture on the Collagen-induced arthritis(CIA) in rats. Methods : Arthritis was induced by intradermal injection of Bovine type II collagen solution into base of tail. Experimental group were divided into 5 groups ; Normal(N) group, Control(C) group, Trachelospermi Caulis high(TH) group, Trachelospermi Caulis low(TL) group, Saline(S) group(n=7 for each group). Normal group was had no management. Control group was injected with Bovine type II collagen solution and taken no treatment. Trachelospermi Caulis high group was injected with Bovine type II collagen solution and taken high-intensity(10mg/kg) herbal-acupuncture treatment on $ST_{36}$. Trachelospermi Caulis low group was injected with Bovine type II collagen solution and taken low-intensity(5mg/kg) herbal-acupuncture treatment on $ST_{36}$. Saline group was injected with Bovine type II collagen solution and taken saline injection on $ST_{36}$. Body weight, paw edema volume and ankle joint thickness were measured during experimental day. On the last experimental day, we analyzed WBC count, TNF-$\alpha$ & IL-$1{\beta}$ concentration, c-fos immunohistochemistry and NADPH-d histochemistry for evaluating the effect of Trachelospermi Caulis herbal-acupuncture. Results : The results were as follows ; 1. In the change of paw edema volume, TH group only has significant difference compared with C group. 2. In the change of ankle joint thickness, TH group only has significant difference compared with C group. 3. In WBC count of serum, TH, TL groups have significant decrease compared with C group. 4. In TNF-$\alpha$ concentration of effusion, TH, TL groups have significant decrease compared with C group. and TH group has significant decrease compared with TL group. 5. In IL-$1{\beta}$ concentration of effusion, TH, TL groups have significant decrease compared with C group. and TH group has significant decrease compared with TL group. 6. In c-fos positive neurons of S1S2(cortex) region, TH, TL, S groups have significant decrease compared with C group. 7. In NADPH-d positive neurons of CPu(caudate putamen) region, TH, TL groups have significant decrease compared with C group. 8. In NADPH-d positive neurons of Tfp(transverse fibers of pons) region, TH, TL, S groups have significant decrease compared with C group. and TH group has significant decrease compared with S group. Conclusions : According to above results, we hope that Trachelospermi Caulis herbal-acupuncture may have the effect that decreases progression and development of CIA. And it can be suggested that Trachelospermi Caulis herbal-acupuncture may reduce the expression of c-fos and NOS.
Background: Current management strategies attempt to diagnose rheumatoid arthritis (RA) at an early stage. Transcription profiling is applied in the search for biomarkers for detecting early-stage disease. Even though gene profiling has been reported using several animal models of RA, most studies were performed after the development of active arthritis, and conducted only on the peripheral blood and joint. Therefore, we investigated gene expression during the initial phase of collagen-induced arthritis (CIA) before the arthritic features developed in the thymus in addition to the peripheral blood and synovium. Methods: For gene expression analysis using cDNA microarray technology, samples of thymus, blood, and synovium were collected from CIA, rats immunized only with type II collagen (Cll), rats immunized only with adjuvant, and unimmunized rats on days 4 and 9 after the first immunization. Arrays were scanned with an Illumina bead array. Results: Of the 21,910 genes in the array, 1,243 genes were differentially expressed at least 2-fold change in various organs of CIA compared to controls. Among the 1,243 genes, 8 encode T-cell receptors (TCRs), including CD3${\zeta}$, CD3${\delta}$, CD3${\varepsilon}$, CD8${\alpha}$, and CD8${\beta}$ genes, which were down-regulated in CIA. The synovium was the organ in which the genes were differentially expressed between CIA and control group, and no difference were found in the thymus and blood. Further, we determined that the differential expression was affected by adjuvant more than Cll. The differential expression of genes as revealed by real-time RT-PCR, was in agreement with the microarray data. Conclusion: This study provides evidence that the genes encoding TCRs including CD3${\zeta}$, CD3${\delta}$, CD3${\varepsilon}$, CD8${\alpha}$, and CD8${\beta}$ genes were down-regulated during the initial phase of CIA in the synovium of CIA. In addition, adjuvant played a greater role in the down-regulation of the CD3 complex compared to CII. Therefore, the down-regulation of TCR gene expression occurred dominantly by adjuvant could be involved in the pathogenesis of the early stage at CIA.
Objectives This study was designed to evaluate the healing effect of Cordyceps Militaris (CM) on collagen II-induced arthritis rats. Methods Sprague-Dawley rats were randomly divided into 6 groups (normal, control, positive control, CM with low/medium/high dosage each). Type II collagen mixed with complete Freund's adjuvant (with 1:1 v/v) was injected subcutaneously, and the mixture was injected in a same manner one week after the first injection to boost arthritis. Arthritis index, paw thickness and von Frey test were conducted to observe physical changes. hematoxylin and eosin (H&E) staining was performed to observe knee cartilage. The levels of messenger RNA (mRNA) expressions of interleukin (IL)-1𝛽, IL-6, tumor necrosis factor-alpha (TNF-𝛼) in spleen were assessed by real-time polymerase chain reaction. Results Rheumatoid arthritis is an autoimmune disease that occurs on multiple joints and can lead to temporary shape change of bones or organ failure in severe cases. Here, we aimed to determine the effect of CM extract on rheumatoid arthritis by measuring paw thickness, arthritis index, conducting von Frey test and H&E staining, and evaluating the level of IL-1𝛽, IL-6, TNF-𝛼. As a result, paw thickness, arthritis index significantly decreased in low concentration group, hind leg became less sensitive in all expermental groups. Also, histological analysis showed that the damage of knee cartilage was prevented in all experimental groups. The level of mRNA of IL-1𝛽, IL-6, and TNF-𝛼 in spleen was analyzed to decide the effectiveness of CM extract. IL-1𝛽 did not show significant change, but IL-6 and TNF-𝛼 showed significant decrease in at least one of the experimental groups. Conclusions CM showed protective effect on knee tissue destruction and improved the physical conditions of the leg involving arthritis. Also, it showed that CM has anti-inflammatory effect on specific cytokines inducing rheumatoid arthritis. In conclusion, this study demonstrated that the therapeutic potential of CM for the treatment rheumatoid arthritis, and set the foundation for the further studies.
Jo, Young-Wook;Yoon, Jong-Hwa;Kim, Kyung-Ho;Lee, Seung-Deok;Kim, Kap-Sung
Journal of Acupuncture Research
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제22권2호
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pp.93-101
/
2005
Objective: We have evaluated UDHA into the joint for its effectiveness on immune responses to CII in the rat CIA. In an attempt to gain further insight into the mode of action of UDHA, we also investigated the effects of UDHA on the incidence and development of arthritis in rat CIA with 2 different regimens: (1) started prior to a primary immunization, (2) started on the day of a primary immunization. Methods : Male rats were immunized with an emulsion of $200\;{\mu}g/100g$ of CII and complete Freund's adjuvant (CFA). The rats were then given intraperitoneal(i.p) stimulation of Ulmus davidiana Planch herbal acupuncture(UDHA) or saline during the experiment. Lymph node cells were obtained from rats 14 days after immunization and cultured in vitro with CII. When compared with rats treated with saline as control, UDHA at doses of more than $20\;{\mu}g/100\;g$ rat once a day for 7 days inhibited the ability of inguinal lymph node cells to produce T cell cytokines interleukin-$1{\beta}$, tumor necrosis $factor-{\alpha}$$(TNF-{\alpha})$. When rats were injected intraperitoneally with SRBC, hemaglutination titers in UD-treated and control rats did not differ significantly when low doses of UD was given to rats. However, i.p injection of UD at doses of more than $10\;{\mu}g/100\;g/day$ for 7 days slightly suppressed antibody production. Results : The present results show that treatment with UDHA can inhibit the onset and development of arthritis and the immune responses to collagen. Conclusion: Therapeutic i.p injection with UD affect the clinical course of the disease and the immune responses to CII.
Objectives: The study was designed to evaluate anti-arthritic activity of Ajmodadi Churna (AC) and its effect on Complete freund's adjuvant (CFA)-induced arthritis in Wistar rats. Methods: Arthritis was induced by injecting 0.2 mL CFA into sub plantar surface of left hind paw. Test sample AC-1 and AC-2, 200 and 400 mg/kg, respectively was given to the animals for 21 consecutive days. The increase in swelling was observed after induction of arthritis. The paw edema was measured on 0, 3, 7, 14 and 21 day using Vernier caliper after the induction of arthritis. The collected blood samples further used for the estimation of red blood cells (RBC), white blood cells (WBC), erythrocytes sedimentation rate (ESR), and hemoglobin (Hb), using hematology analyzer. Serum concentration of IL-6 and TNF-α were also measured using rat ELISA kits. Results: Results showed that a significant reduction in paw edema was observed in AC-2 treated rats. The paw edema was restored on day 21 was 4.48 mm for AC-2, which is near to the control group. The arthritis score in treated rats was found to be considerably lower than in the control group i.e. 0.83 for AC-2 and 1.50 for AC-1. A decrease in levels of RBC and hemoglobin were observed in arthritic rats. Inflammation was significantly reduced and serum levels of IL-6 and TNF-α were lowered after treatment with the test drug. Conclusion: It can be concluded from the study that AC possess significant anti-arthritic activity. Furthermore, this condition was linked to a reduction in abnormal humoral immune responses.
Objectives : To investigate the analgesic effect and its cholinergic mechanism of electroacupuncture(EA) in the rat model of collagen-induced arthritis(CIA). Methods : Immunization of male Sprague-Dawley rats with bovine typeII (CII) collagen emulsified in Freund's incomplete adjuvant, followed by a booster injection 14 days later, leads to development of arthritis in more than 70% of rats by 21 days postinjection. After three weeks of first immunization, EA stimulation(2 Hz, 0.07 mA, 0.3 ms) was delivered into Jogsamni($ST_{36}$) for 30 minutes. Analgesic effect was evaluated by tail flick latency(TFL). We compared the analgesic effect of EA with TFLs between pretreatment of normal saline and pretreatment of Atropine (1 mg/kg, intraperitoneal) and Neostigmine ($100{\mu}g/kg$, intraperitoneal) in CIA. Results : 1. TFLs were gradually decreased in CIA as increasing severity of arthritis. 2. Jogsamni($ST_{36}$) EA stimulation in CIA increased TFLs and the effect lasted for 60 minutes. 3. Increased TFLs with Jogsamni($ST_{36}$) EA stimulation were inhibited with pretreatment of atropine in CIA 4. Increased TFLs with Jogsamni($ST_{36}$) EA stimulation did not show an obvious synergistic effect with pretreatment of neostigmine in CIA. Conclusions: Jogsamni($ST_{36}$) EA showed analgesic effects in CIA. The analgesic effects of Jogsamni($ST_{36}$) EA were inhibited by atropine pretreatment and combined application of Jogsamni(ST36) EA and neostigmine did not show an synergistic effect. These observations suggest that intrinsic muscarinic cholinergic pathways represent an important modulating system in pain perception of inflammatory pain in CIA It is suggested that, the active mechanism of analgesic effect in EA may involve the release of acetylcholine in the spinal cord.
Objectives This study was carried out to find the effects of Gami-cheongyulsaseub-tang (hereinafter referred to GCST) on the inhibition of zymosan-induced pain in rats and collagen II-induced arthritis (CIA) in DBA/1J mouse. Methods As an acute inflammatory pain model, peripheral inflammation was induced by intraplantar injection of zymosan into the right hind paw in rats and then the hyperalgesia and pain regulating factors in spinal cord were analyzed. As a chronic inflammation model, the mixture of collagen II and complete Freund's adjuvant was treated into mice to establish rheumatoid arthritis and then body weight, thickness of hind paw, pathological change of spleen, immunological rheumatoid factor (IgG1, IgG2a, IgG2b, IgM and anti-collagen II), pro-inflammatory cytokines, and bone injury were analyzed. Results In the acute inflammatory pain model, GCST significantly inhibited the thermal and mechanical hyperalgesia and the pain regulating factors, including Fos, CD11b, PKA and PKC, in the spinal cord with a dose-dependent manner. In the chronic rheumatoid arthritis model, GCST administration decreased arthritic index and paw edema as compared with CIA control group. In particular, GCST reduced significantly the serum levels of total IgG2a, IgG2b, IgM, and specific anti-collagen II, but not total IgG1. GCST also resulted in the attenuation of bone injury and spleen enlargement/adhesion in CIA mice. Moreover, the secretion of pro-inflammatory cytokines TNF-${\alpha}$ and IL-$1{\beta}$ in CIA mice was significantly reduced by GCST in a dose-dependent manner. Conclusions Comparison of the results in this study showed that GCST had anti-nociceptive and immunomodulatory effects. These data imply that GCST can be used as an effective drug for not only rheumatoid arthritic pain but also other auto-immune diseases.
Collagen 유발(誘發) 관절염(關節炎) 동물모델을 이용하여 족삼리(足三里) (ST36) 봉독약월(蜂毒藥鉞)의 진통효과(鎭痛效果)와 그 기전(機轉) 을 관찰하기 위하여 TFL을 지표로 하여 통증 역치의 변화와 naloxone 및 yohimbine으로 전처(前處) 치한(置) 후 (後) 봉독약침(蜂毒藥鍼)의 진통효과(鎭痛效果)를 관찰한 결과 다음과 같은 결론을 얻었다. 1. Collagen 유발(誘發) 관절염(關節炎)의 rat에서 TFL 통증 역치는 1, 2, 3 및 4주로 갈수록 지속적 감소를 나타내었다. 2. 1회와 2주간 봉독약철(蜂毒藥鐵)은 collagen 유발(誘發) 관절염(關節炎) rat의 통증역치를 각각 증가시켰다. 3. CIA rat에 opioid receptor antagonist인 naloxone을 투여한 후 봉독약침(蜂毒藥鍼)을 처치한 결과 진통효과(鎭痛效果)가 유지되는 것으로 보아 봉독약광(蜂毒藥鑛)의 진통기전(鎭痛機轉)은 아편양 기전(機轉)과 관련이 없는 것으로 사료된다. 4. CIA rat에 ${\alpha}2$-adrenoceptor antagonist인 ohimbine을 투여한 후 봉독약침(蜂毒藥鍼)을 처치한 결과 진통효과(鎭痛效果)가 길항(拮抗)되는 것으로 보아, 봉독약침(蜂毒藥鍼)의 기전(機轉)은 ${\alpha}2$-adrenoceptor에 의하여 매개는 것으로 사료된다.
The purpose of this study was to observe the effects of Achyranthes Radix(AR) and electroacupuncture(EA) in rats with rheumatoid arthritis induced by type II collagen for 28 days. Control group was daily administered 0.9% NaCl 0.5 $m{\ell}$, Group I was daily administered 0.9% NaCl 0.5 $m{\ell}$ to arthritic rats, Group II was orally administered with Achyranthes Radix 500 mg/kg 0.5 $m{\ell}$ to arthritic rats. Group III was given 2 Hz EA of chok samni acupoint(ST36) in the test group for 30 min/days to arthritic rats. Group IV was daily orally administered with Achyranthes Radix 500 mg/kg 0.5 $m{\ell}$ and 2 Hz EA of chok samni acupoint(ST36) in the test group for 30 min/days to arthritic rats. This studies have been designed to evaluate the hind paw edema, assessment of arthritis indices, analgetic effects by analysis of blood chemistry(WBC, CRP, ALP, AST). In each group, histologic observations, Safranin O-fast green stain were observed and analyzed. The following results were obtained. Group II, III, IV were significantly decreased arthritis indices and the rate of paw edema compared with Group I . Especially group IV was the most significantly decreased. The WBC, CRP, AST, ALT was that Group II, III, IV were significantly decreased compared with Group I . In conclusion, Achyranthes Radix and Ea contribute to the improvement of blood chemistry and change in safranin O-fast green by knee joint of arthritic rats.
Purpose: This study examined the effects of swimming exercise and Achyranthes Radix extracts on the inflammatory and behavioral responses in type II collagen-induced arthritic rats for 28 days. Methods: Sprague-Dawley rats were allocated randomly to one of the following four groups: only type II collageninduced (group Ⅰ), application of swimming exercise after type II collagen-induced (group II), application of Achyranthes Radix ointment after type II collagen-induced (group III), application of swimming exercise and Achyranthes Radix ointment after type II collagen-induced (group IV). Arthritis was established in SD rats by an intradermal injection of Chick type II collagen plus incomplete Freund's adjuvant at the base of the tail of the animals. The swimming exercise program consisted of a 25 min swimming session/day with a load corresponding to 5.5% of the weight bearing, three days/week for four weeks. The Achyranthes Radix ointment (0.1g) was applied twice a day for five days. The changes in behavior, H & E stain, and cyclooxygenase-2 (COX-2) level in the knee joint were assessed. Results: The gross and histological examination, after RA induction showed reddening, edema and erythema. The H & E stain revealed the destruction of articular cartilage, bony erosion and the infiltration of inflammatory cells after RA induction. The mechanical allodynia test results were significantly higher in group I than in groups II, III and IV (p<0.01). The immunohistochemistrical response of COX-2 in the knee joint showed that groups II, III, IV had a lower response effect than group I. Conclusion: Swimming exercise training and Achyranthes Radix ointment decreased the inflammatory responses and enhanced the behavioral responses in the arthritic rats.
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