Protective effects of sigma 1 receptor agonist PRE084 on 2,4,6-trinitrobenzene sulfonic acid–induced experimental colitis in mice

Hyun Il Seo,권성춘,Jae Young Kwak 대한외과학회 2022 Annals of Surgical Treatment and Research(ASRT) Vol.103 No.3

Purpose: We aimed to investigate the protective effect of sigma 1 receptor agonist and antagonist, PRE084 and BD1047, respectively, on 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis in mice. Methods: Thirty male ICR mice were randomly divided into 5 groups: control, 50% ethanol, colitis, PRE084 + colitis, and combined (PRE084 + BD1047 + colitis). Colitis was induced by intrarectal administration of TNBS. PRE084 and BD1047 were injected daily, starting 3 days before colitis induction. Distal colon tissue was excised for histopathological evaluation, and levels of glutathione (GSH), superoxide dismutase (SOD), myeloperoxidase (MPO), and lipid peroxidation were determined. Results: Colitis caused weight loss, mucosal damage, upregulation of tumor necrosis factor-α, interleukin (IL)-1β, IL- 6, MPO, and thiobarbituric acid reactive substance activities, and downregulation of GSH and SOD activities. These changes caused by TNBS-induced colitis were significantly ameliorated by PRE084 pretreatment. However, the combined pretreatment with BD1047 significantly attenuated the protective effect of PRE084, thereby reverting to the colitis-induced state. Conclusion: We conclude that the sigma 1 receptor agonist PRE084 exhibits significant protective effects against TNBS- induced colitis, which appears to be at least partly mediated by the inhibition of inflammation and oxidative stress, and enhancement of antioxidant properties. Collectively, these results suggest that PRE084 might be an effective drug for the treatment of ulcerative colitis.

합곡의 구진약침이 흰쥐의 염증성 대장염에 미치는 치료 효과

송재수 ( Jae Soo Song ),양범식 ( Beum Sik Yang ),김선영 ( Sun Young Kim ),홍정아 ( Jeong A Hong ),송정방 ( Jeong Bang Song ),김경식 ( Kyung Sik Kim ),김재효 ( Jae Hyo Kim ),권오상 ( Oh Sang Kwon ),손인철 ( In Chul Sohn ) 대한경락경혈학회 2007 Korean Journal of Acupuncture Vol.24 No.2

Objectives: Ulcerative colitis or Crohn`s disease has been recognized as Ha-ri (下痢) or Jang-Byok (腸벽) in Korean oriental medicine. A purpose of the present study is to investigate the anti-inflammatory effect of moxi-tar herbal acupuncture at LI4 (HapGok) on 2,4,6-trinitrobenzene sulphonic acid (TNBS) induced colitis in rats and further elucidate the possibility of herbal acupuncture on ulcerative colitis which is chronic inflammatory disease of the gastrointestinal tract. Methods: Sprague-Dawley rats, weighing 170~190 g, were subjected to intrarectal injection of either saline (300 ㎕, 500 ㎕) for a control or 2,4,6-trinitrobenzene sulphonic acid (TNBS) (300 ㎕, 500 ㎕) for a colitis. Moxi-tar herbal acupuncture at bilateral LI4 was used as the treatment. A volume 0.4 ㎖of moxi-tar (20 ㎎/㎖) were subcutaneously injected to the LI4 just after the secondary injection time of TNBS in rats. To study the effects of Moxi-tar acupuncture in LI4, body weight, RBC count, WBC count, total protein, Paw edema rate, rate of protein leakage into CMC-pouch fluid, IgG levels and IgM levels were observed. Results: Moxi-tar acupuncture in LI4 on TNBS-induced colitis inhibited the body weight lose rate but not effect RBC, WBC count. In addition, it inhibited the reduction of total protein concentration, paw edema, rate of protein leakage into CMC-pouch fluid, IgG levels and IgM levels. Conclusions: It is suggested that moxi-tar herbal acupuncture at LI4 helps to recover TNBS-induced colitis and plays an important role for an treatment of the irritable bowel syndrome (IBS).

위장관 : 급성 TNBS-유발 대장염에서 동종 골수이식에 의한 치료 효과

맹이소 ( Lee So Maeng ),장은덕 ( Eun Duck Chang ),채현석 ( Hiun Suk Chae ),김진수 ( Jin Soo Kim ),민정요 ( Jeong Yo Min ),손혜숙 ( Hye Sook Sohn ),노상영 ( Sang Young Rho ),김형근 ( Hyung Keun Kim ),조영석 ( Young Suk Cho ),최규용 대한소화기학회 2009 대한소화기학회지 Vol.54 No.1

목적: 골수에서 유래한 세포들은 여러 병적인 환경에서 조직을 유지하는 데 기여한다. 저자들은 흰쥐의 실험 대장염에서 골수 이식 시 대장염의 치료에 대해 골수유래세포들의 역할을 알아보기 위해 연구를 시행하였다. 대상 및 방법: 실험에 사용한 흰쥐는 3개의 군으로 나누어 대조군 (50% ethanol), 2,4,6-trinitrobenzene sulfinic aicd (TNBS군) 장염군, TNBS+골수이식군(BMT군)으로 하였다. 장염을 유발하기 위해 50% 알코올에 녹인 TNBS (5.0 mg/마리)를 일주일에 한 번씩 2주 동안 항문을 통해 투여하였다. 동종 골수 이식은 TNBS 투여 3주 전에 green fluorescence protein (GFP)를 지닌 수컷 유전자 도입 쥐의 골수 세포를 야생형 생쥐의 꼬리 정맥에 투여하였다. 모든 동물은 TNBS 투여 후 일주일 후에 희생하여 대장을 추출하였다. 골수이식이 되었는지를 확인하기 위해 GFP에 대한 면역조직화학검사를 시행하였고 상피하 근육섬유모세포의 존재를 확인하기 위해 vimentin과 α-SMA에 대한 면역조직검사를 시행하였다. 결과: 장염의 정도는 TNBS군에서 가장 심하였고 골수이식에 의해 유의하게 감소하였다(p<0.05). GFP 양성 세포는 주로 선와의 니쉐 부위에서 염색되었으며 골수 이식군에서만 양성이었다. 근육섬유세포에 대한 vimentin, α-SMA에 대한 염색도 주로 선와 니쉐에서 양성이었고 대조군이나 TNBS군에 비해 골수 이식군에서 많았다. 결론: 흰쥐의 급성 대장염 치료에 골수이식은 효과적이며 골수유래세포는 선와니쉐 부위에서 근육섬유세포로 분화되어 장염의 치료에 관여한다. Background/Aims: Bone marrow-derived cells (BMDC) contribute to tissue maintenance under many kinds of pathologic conditions. We carried out a study to see how BMDC play a role in the treatment of experimental murine colitis. Methods: We divided the animals into 3 groups and treated them with 50% ethanol (control group), 2,4,6-trinitrobenzene sulfinic acid colitis (TNBS group), and TNBS+bone marrow transplant (BMT group). To induce colitis, TNBS (5.0 mg/mouse) dissolved in 50% ethanol was injected into anus weekly for two weeks. Bone marrow transplantations were performed using bone marrow of male transgenic mouse (donor) with green fluoresence protein (GFP) into female wild type mouse (recipient) three weeks before TNBS instillation. All animals were sacrificed, and colons were extracted one week after the last TNBS instillation. We measured microscopic scores of mucosal injury and investigated the GFP expression for bone marrow engraftment. The immunostaining of vimentin and α-smooth muscle actin (α-SMA) for myofibroblasts was performed. Results: The score of mucosal injury in the TNBS group was much more severe than those in control, and reduced significantly by BMT (p<0.05). GFP-positive cells were almost deposited in pericryptal niche of BMT group but not at all in both control and TNBS group. Most of myofibroblasts stained with both vimentin and SMA also infiltrated into pericryptal niche. But, the number of myofibroblasts stained with vimentin and SMA in both control and TNBS group was smaller than that in BMT group. Conclusions: BMDC deposited on pericryptal niche might have a significant role in repairing acute experimental murine colitis. (Korean J Gastroenterol 2009;54:20-27)

흰쥐의 Trinitrobenzene Sulfonic Acid ( TNBS ) 유발성 대장염에서 Flavonoids 계 Eupatilin 유도체 DA - 6034 의 효과

장동경(Dong Kyung Chang),진영주(Young Joo Chin),정현채(Hyun Chae Jung),송인성(In Sung Song),김정룡(Chung Yong Kim),손미원(Mi Won Son),유무희(Moo Hi Yoo) 대한내과학회 1998 대한내과학회지 Vol.55 No.3

N/A Objective: In traditional medicine, Artemisia capillaris has been used for treatment of chronic diarrhea. Previously we found Artemisia capillaris had an effect on rats with TNBS-induced colitis. Eupatilin, a kind of fla- vonoids, may be a probable effective component. To evaluate the effect of a eupatilin derivative compound DA-6034 on the rat with TNBS-induced colitis, we perfomed this study, Methods: Colitis was induced with 1ml of 50 mg/ml TNBS mixed with 60 % ethanol (vol/vol) in Sprague Dawley rats. From the next day, 1ml methylcellulose, 1 mg/kg prednisolone, 0.3 or 3 mg/kg of DA-6017 and DA-6034 were administered through once daily for 2 weeks. At 2days, lweek, and 2weeks later, we evaluated the effect by gross damage score (0-10) and measured myeloperoxidase, PGE2, and L from the damaged mucosa. Results: The mean gross damage scores of prednisolone end 3 mg/kg of DA groups were significantly lower than that of a placebo group at 2weeks(0.8, 0.9 vs. 4.0, p<0.05). Myeloperoxidase activities also seemed to be lower in those effective groups but were not statistically significant. LTB4 levels were lower in prednisolone and, 0.3 and 3 mg/kg of DA groups than in a placebo group at 2weeks(7.91, 7.23, and 7.13 vs, 13.90 ng/mg protein, p<0.05), PGE2 levels were decreased in prednisolone and 0.3 mg/kg of DA-6034 groups at 2days. DA17 showed no effects. Conclusions: Eupatilin derivative compound, DA-6034 was effective in rats with TNBS-induced colitis. In that LTE4 leve1 is lowered with some decrease of PGE2 level, this agent probably has an inhibitory effect on arachidonic acid metabolism.

Anti-Inflammatory Effect of Crude Momordica charantia L. Extract on 2,4,6-Trinitrobenzene Sulfonic Acid-Induced Colitis Model in Rat and the Bioaccessibility of its Carotenoid Content

Nalan Gulsen Unal,Aysegul Kozak,Sibel Karakaya,Nevin Oruc,Burcu Barutc¸uoglu,Cagdas Aktan,Murat Sezak,Ahmet Omer Ozutemiz 한국식품영양과학회 2020 Journal of medicinal food Vol.23 No.6

Momordica charantia L., known as bitter melon (BM), is a plant that belongs to the family Cucurbitaceae. Aims of this study are to investigate the anti-inflammatory effect of crude BM extract on 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis model in rat. It was also aimed to determine the content and bioaccessibility of carotenoids of BM. BM was purchased from local markets in Izmir, Turkey. Fruits of BM were lyophilized, powdered, and used in the experiment. Carotenoids were determined by high-performance liquid chromatography. To determine the bioaccessibility of β-carotene, in vitro digestion was performed. Wistar albino rats were divided into four groups: group A (BM+TNBS), group B (BM), group C (TNBS), and group D (control). BM solution was given 300 mg/(kg·day) for 6 weeks orally. Colitis was induced by 0.25 mL of a solution containing 100 mg/kg 5% (w/v) TNBS in 50% ethanol (w/v) intrarectally after 6 weeks. After sacrification, macroscopic and microscopic evaluations were performed. Myeloperoxidase, cytokines levels (interleukin-17 [IL-17], TNF-alpha, and interleukin-10 [IL-10]) were measured in serum and colonic samples by ELISA test. Institutional Animal Ethics Committee approval was obtained. Total carotenoid content of BM was determined 11.7 mg/g dry weight as β-carotene equivalents. Bioaccessibility of total carotenoids was determined as 2.1% with in vitro digestion. Pretreatment with crude BM extract significantly reduced weight loss, macroscopic, and microscopic colitis damages in colonic samples (P = .000), (P = .015), and (P = .026), respectively. Serum anti-inflammatory cytokine IL-10 increased significantly in both treatment groups (P = .000). BM is a rich source of carotenoids, but the bioaccessibility of its carotenoids is low. This study displays that BM has protective anti-inflammatory effects on TNBS-induced colitis.

Rebamipide Protects TNBS Induced Colonic Damage Through Down-regulation of NF-κB Activation and Induction of Heme Oxygenase-1 Expression

오재민(Jae Min Oh),이정래(Jeong Re Lee),권영미(Young-Mi Kwon),김유림(Yu Rim Kim),김경숙(Kyoung Suk Kim),최민규( Min Kyu Choi),정연태(Yeun Tai Chung) 대한해부학회 2005 Anatomy & Cell Biology Vol.38 No.1

크론병은 proinflammatory cytokine들이 중요한 역할을 하는 만성 염증성 대장 질환이다. Rebamipide는 위궤양 치료제로 개발된 약제로 항염증 작용이 있다고 알려져 있으나 크론병 환자에서 그 작용기전은 정확하게 알려져 있지 않다. 따라서 우리는 크론병 동물모델인 TNBS에 의해 유도된 만성 대장염 모델에서 rebamipide의 항 염증 기전을 조사하였다. TNBS에 의해 유도된 만성 대장염에서 rebamipide는 MPO 활성 및 병리조직학적 증상들을 현저히 개선하였다. Rebamipide는 대장상피세포주인 HT-29세포에서 TNF-α에 의한 Il-8 분비, ICAM-1 발현, NF-κB 활성 등을 억제하였다. 또한 rebamipide는 HT-29세포에서 항염증작용이 있다고 알려진 heme oxygenase-1 (HO-1)의 발현을 증가시켰다. HO-1의 유도제인 copper protoporphyrin IX (CoPPIX)는 HT-29세포에서 NF-κB 활성 등을 억제하였고 TNBS에 의해 유도된 만성 대장염에서의 rebamipide의 효과를 재현하였다. 이상의 결과로 Rebamipide는 HO-1의 유도를 통해서 크론병의 치료에 있어 중요한 치료제가 될 수 있다고 생각하였고 HO-1은 크론병의 치료에 중요한 표적물질이 될 수 있다고 생각하였다. Crohn’s disease is characterized by a chronic relapsing inflammation of the bowel in which proinflammatory cytokines play an important role. Rebamipide is an anti-gastric ulcer drug with anti-inflammatory properties in vivo and in vitro. The effects of rebamipide on Crohn’s disease have not been carefully evaluated. This study investigated the potential of rebamipide to protect Crohn’s disease using a murine model of colitis induced by trinitrobenzene sulfonic acid (TNBS). Rebamipide dramatically improved histopathological symptom involving myeloperoxidase (MPO) activation and increase of microscopic damage score in TNBS induced colitis. Rebamipide suppressed IL-8 secretion, ICAM-1 induction and nuclear factor-κB (NF-κB) activation by TNF-α and induced heme oxygenase-1 (HO-1) in HT-29 cells. HO-1 inducer cobalt protoporphyrin IX (CoPPIX) suppressed NF-κB activation by TNF-α in HT-29 cells like rebamipide, and mimicked the protective effects of rebamipide on TNBS induced colitis. This suggests that rebamipide exerts anti-inflammatory effects by down-regulating NF-κB activity via inducting HO-1 expression. In conclusion, this study suggests that rebamipide represents a potential therapeutic agent and HO-1 is an important therapeutic target for the treatment of Crohn’s disease.

In vitro and in vivo inhibitory activity of 6-amino-2,4,5-trimethylpyridin-3-ols against inflammatory bowel disease

Banskota, S.,Kang, H.e.,Kim, D.G.,Park, S.W.,Jang, H.,Karmacharya, U.,Jeong, B.S.,Kim, J.A.,Nam, T.g. Pergamon Press 2016 Bioorganic & medicinal chemistry letters Vol.26 No.19

Although the pathogenesis of inflammatory bowel disease (IBD) is complex, attachment and infiltration of leukocytes to gut epithelium induced by pro-inflammatory cytokines such as tumor necrosis factor-α (TNF-α) represents the initial step of inflammation in IBD. Previously, we have reported that some 6-amino-2,4,5-trimethylpyridin-3-ols have significant levels of antiangiogenic activity via PI3K inhibition. Based on the reports that angiogenesis is involved in the aggravation of IBD and that PI3K is a potential target for IBD therapy, we investigated whether the scaffold has inhibitory activity against in vitro and in vivo models of colitis. Many analogues showed >80% inhibition against TNF-α-induced monocyte adhesion to colon epithelial cells at 1μM. Compound 8m showed IC<SUB>50</SUB>=0.19μM, which is about five orders of magnitude better than that of 5-aminosalicylic acid (5-ASA, IC<SUB>50</SUB>=18.1mM), a positive control. In a rat model of 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis, orally administered 8m dramatically ameliorated TNBS-induced colon inflammation. It was demonstrated by a high level of suppression in myeloperoxidase (MPO), a surrogate marker of colitis, as well as almost perfect recovery of colon and body weights in a dose-dependent manner. Compared to sulfasalazine, a prodrug of 5-ASA, compound 8m showed >300-fold better efficacy in those parameters. Taken together, 6-amino-2,4,5-trimethylpyridin-3-ols can provide a novel platform for anti-IBD drug discovery.

Lipophilic modification enhances anti-colitic properties of rosmarinic acid by potentiating its HIF-prolyl hydroxylases inhibitory activity

Jeong, S.,Park, H.,Hong, S.,Yum, S.,Kim, W.,Jung, Y. North-Holland ; Elsevier Science Ltd 2015 european journal of pharmacology Vol.747 No.-

Inhibition of hypoxia inducible factor-prolyl hydroxylase-2 (HPH), leading to activation of hypoxia inducible factor (HIF)-1 is a potential therapeutic strategy for the treatment of colitis. Rosmarinic acid (RA), an ester of caffeic acid and 3,4-dihydroxyphenyllactic acid is a naturally occurring polyphenolic compound with two catechols, a or inhibition of HPH. To improve accessibility of highly hydrophilic RA to HPH, an intracellular target, RA was chemically modified to decrease hydrophilicity. Of the less-hydrophilic derivatives, rosmarinic acid methyl ester (RAME) most potently inhibited HPH. Accordingly, RAME prevented hydroxylation of HIF-1α and consequently stabilized HIF-1α protein in cells. RAME inhibition of HPH and induction of HIF-1α were diminished by elevated doses of the required factors of HPH, 2-ketoglutarate and ascorbate. RAME induction of HIF-1α led to activation of an ulcer healing pathway, HIF-1-vascular endothelial growth factor (VEGF), in human colon carcinoma cells. RAME administered rectally ameliorated TNBS-induced rat colitis and substantially decreased the levels of pro-inflammatory mediators in the inflamed colonic tissue. In parallel with the cellular effects of RAME, RAME up-regulated HIF-1α and VEGF in the inflamed colonic tissue. Thus, lipophilic modification of RA improves its ability to inhibit HPH, leading to activation of the HIF-1-VEGF pathway. RAME, a lipophilic RA derivative, may exert anti-colitic effects via activation of the ulcer healing pathway.

Antiinflammatory activity of an herbal preparation (HemoHIM) in rats

Jo, Sung Kee,Lee, Hae June,Kim, Se Ra,Kim, Jong Choon,Bae, Chun Sik,Jung, Uhee,Park, Hae Ran,Jang, Jong Sik,Kim, Sung Ho Heyden & Son 2007 Phytotherapy research Vol.21 No.7

<P>This study evaluated a new herbal preparation, HemoHIM, for its antiinflammatory activity against carrageenan-induced edema, the formation of granulation tissues by cotton pellet and experimental colitis by 2,4,6-trinitrobenzene sulfonic acid (TNBS). The HemoHIM was prepared by adding its ethanol-insoluble polysaccharide fraction to the total water extract of Angelica Radix, Cnidii Rhizoma and Paeonia Radix. The preparation (4 mg of solids/mL of drinking water, p.o., 50–100 mg/kg of body weight, i.p.) produced a dose-related inhibition of carrageenan-induced paw edema and cotton pellet-induced granuloma in rats. In addition, HemoHIM also reduced the degree of TNBS-induced colitis and improved the gross and histological changes such as thickening, dilatation, ulceration, and infiltration by polymorphonuclear leukocytes and multiple erosive lesions. These results demonstrate that the HemoHIM has a potent antiinflammatory effect. Copyright © 2007 John Wiley & Sons, Ltd.</P>