• Korean Journal of Veterinary Research
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• v.51 no.2
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• pp.83-91
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• 2011

In this study, immunohistochemistry was used to examine the changes in the density of colonic endocrine cells - argyrophil and argentaffin cells, chromogranin A (CGA), serotonin, somatostatin and glucagon-containing cells in trinitrobenzene sulfonic acid (TNBS)-induced rat colitis. Ulcerative colitis was induced by the instillation of 10 mg of TNBS into the colonic lumen through the anus. To confirm the inducement of ulcerative colitis, the macroscopic and microscopic scores as well as the colonic myeloperoxidase (MPO) activities were monitored for 8 days after TNBS instillation in the colonic lumens. In addition, the number of argyrophil and argentaffin cells, CGA, serotonin, somatostatin and glucagon-immunoreactive cells were counted in the colonic mucosa, respectively. After TNBS instillation into the lumen of the colon from the anus in rats, increases in macroscopic and microscopic scores in the colon tissues were observed along with increases in the colonic MPO activities. Therefore, ulcerative colitis was relatively well induced by the TNBS instillations. Marked decreases in the number of colonic endocrine cells were detected in the TNBS-treated animal compared to the sham control. These results suggest that colonic endocrine cells were also disrupted by TNBS-induced ulcerative colitis.

• Korean Journal of Acupuncture
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• v.25 no.2
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• pp.159-177
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• 2008

Objectives : Crohn's disease is a severe chronic inflammation that is treated mainly by immunosuppression, which often has serious side effects. There is need to develop new therapeutic methods or drugs that have few side effects in order to treat this disease. Acupuncture with Moxi-tar at Cheonchu (ST25) has anti-inflammatory properties, but the mechanism of its anti-inflammatory actions is unclear. We investigated the protective effects and speculated the mechanisms of acupuncture with Moxi-tar at ST25 on trinitrobenzene sulfonic acid (TNBS) induced colitis in mice which is a well known Crohn's disease animal model. Methods : 5 % TNBS was treated at day 1 and day 7 into rectum of mice. To investigate therapeutic effects of acupuncture with Moxi-tar at ST25, acupuncture was carried out on day 3, and day 6. For the data analysis, we observed macroscopic and microscopic findings of the colon. Weight and width of the colon, degree of damage, changes of body weight, and myeloperoxygenase (MPO) activity were checked. For analysing protein expression, we carried out immunohistochemical staining and Western blot. For analysing mRNA expression, RT-PCR was carried out. Results : TNBS induced damages on the colon of mice, while acupuncture of Moxi-tar at ST25 suppressed TNBS mediated damages similar to those on the colons of mice in the control (not treated with TNBS) group. The average body weight of TNBS treated mice (77.4%) was decreased compared with that of the control mice (105%), and acupuncture with Moxi-tar at ST25 suppressed the loss of body weight caused by TNBS (from 77.4% to 95.3%). TNBS induced infiltration of immune cells in all layers of the colon while acupuncture with Moxi-tar at ST25 suppressed infiltration of immune cells caused by TNBS. Furthermore, acupunctured with Moxi-tar at ST25 suppressed macro-, micro- colonic damages caused by TNBS. Acupunctured with Moxi-tar at ST25 dramatically improved the clinical and histopathological symptoms such as the increase in weight of the distal colon and the MPO activity in TNBS-induced colitis. Acupuncture with Moxi-tar at ST25 down-regulated the nuclear transcription factor kappa B ($NF-{\kappa}B$) activity and suppressed tumor necrosis factor-a (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-1${\beta}$), and intracellular adhesion molecule-1 (ICAM-1) expressions caused by TNBS. Conclusions : Acupuncture with Moxi-tar at ST25 helps recovery from the TNBS-induced colonic damage by down-regulation of $NF-{\kappa}B$ activity and suppressing of TNF-${\alpha}$, IL-1${\beta}$, and ICAM-1 expressions. This may be an important method for the treatment of Crohn's disease.

• Journal of Microbiology and Biotechnology
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• v.28 no.1
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• pp.175-179
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• 2018

In a preliminary study, Lactobacillus plantarum C29 was found to suppress 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis in mice. Therefore, to understand whether an anti-colitic probiotic C29 could attenuate memory impairment, we examined the effects of C29 on TNBS-induced memory impairment in mice. Orally administered Lactobacillus plantarum C29 attenuated TNBS-induced memory impairment in mice in the Y-maze, noble object, and passive avoidance task tests. C29 treatment increased TNBS-suppressed hippocampal brain-derived neurotrophic factor expression and inhibited TNBS-induced hippocampal NF-${\kappa}B$ activation and blood LPS levels. Moreover, C29 restored the TNBS-disturbed gut microbiota composition. These findings suggest that C29 can alleviate memory impairment presumably by restoring the gut microbiota composition.

• Korean Journal of Acupuncture
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• v.25 no.3
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• pp.147-166
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• 2008

Objectives : Crohn's disease (CD) is characterized by a chronic relapsing inflammation of the bowel in which proinflammatory cytokines play an important perpetuating role. Methods : Mice (preventive animal model of gliotoxin) were treated with 5 % 2,4,6-trinitrobenzenesulfonic acid (TNBS) at day 1 and day 7. To investigate preventive effects of acupuncture with Gujin at $LI_{11}$, acupuncture was carried out at day -1, day 1, day 3. And, to investigate therapeutic effects, acupuncture with Gujin was carried out at day 3, day 5, day 7. For the data analysis, we checked weight and width of colon, diarrhea, edema, survival rate, changes of body weight, and myeloperoxygenase (MPO) activity. For analysing protein expression, we carried out immunohistochemical staining and Western blot and we analyzed mRNA expression by RT-PCR. Results : Colon of TNBS treated mice was erosive and shortening compared with the colon of control mice and induced damages of colon epithelial cell layer and induced infiltration of immune cells in all layer of colon. Acupuncture of gujin at $LI_{11}$ in preventive mode suppressed macorscopic damages such as erosive and shortening of colon by TNBS and damages of intestinal epithelial cells and infiltration of immune cells in the colon. The average weight of 5 cm distal colon was increased in TNBS treated mice (758${\mu}g$) compared with in control mice (112${\mu}g$) and width of distal colon was also increased in TNBS treated mice (4.9mm) compared with in control mice (1.3mm). Acupuncture with Gujin at $LI_{11}$ in preventive and therapeutic mode suppressed increase of colon weight and width by TNBS. TNBS induced edema of colon and diarrhea and Acupunctured with Gujin at $LI_{11}$ in preventive and therapeutic mode ameliorated these symptom by TNBS. In preventive and therapeutic mode, the effects of acupuncture with Gujin at $LI_{11}$ were increasing the motility, suppressing body weight decreasing, suppressing MPO activity, reducing expressing of TNF-${\alpha}$, IL-1b, and ICAM-1 in colon compared with that by TNBS Conclusions : This study demonstrates that acupuncture with Gujin at $LI_{11}$ represents a potential therapeutic method of Crohn's disease.

• Korean Journal of Acupuncture
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• v.23 no.4
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• pp.147-156
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• 2006

Objectives : The purpose of the present study is to evaluate the effects of herbal acupuncture with Ginseng Radix for the treatment to intestinal disease in the rat with 2, 4, 6-trinitrobenzenesulfonic acid (TNBS) induced colitis. Methods : All animals were subjected to the injection of saline $(300{\mu}{\ell},\;500{\mu}{\ell})$ for a study control or TNBS $(300{\mu}{\ell},\;500{\mu}{\ell})$ into the lumen of the colon, 8cm proximal to the anus through the intestine. Ginseng Radix herbal acupuncture ($20mg/m{\ell},\;0.4m{\ell}$) were injected to the both $Hapgok(LI_4)$ acupoints at 2nd injection of TNBS in rats. Thus, the body weight, RBC count, WBC count, total protein, IgG levels and IgM levels were observed to study the effects of Ginseng Radix herbal acupuncture. Results : Ginseng Radix herbal acupuncture on $Hapgok(LI_4)$ for TNBS-induced colitis inhibited the body weight loss rate but did not affect RBC and WBC counts. Furthermore, it inhibited the reduction of total protein concentration and serum IgG and IgM levels in TNBS induced colitis were recovered. Conclusions : Herbal acupuncture with Ginseng Radix helps recover the TNBS-induced colonic damage and may be an important method for treatment of the colitis.

• Korean Journal of Acupuncture
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• v.26 no.2
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• pp.91-108
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• 2009

Objectives: Single colorectal instillation of trinitrobenzenesulphonic acid (TNBS) causes a dose-dependent increase of visceral motor response (VMR) and severity of inflammation. In this study we compared the effects of electroacupuncture in the different acupoints in the acute colitis induced by TNBS intracolonic injection in rats. Methods: In Male Sprague-Dawley rats, weighing $250{\sim}400g$, a single colorectal administration of TNBS 5mg/kg and 50% ethanol under isoflurane anaesthesia after an overnight fast. Electrodes for electromyography (EMG) recording were stitched into the external oblique musculature under general anesthesia. Acupoints of LI4, ST25, or ST36 were stimulated by electroacupuncture, respectively. The balloon was inserted intra-anally and visceral motor response (VMR) to colorectal distensioin (CRD) was quantified with an EMG recording system. Results: At an observation of the visceral hyperalgesia in the day-time series, the visceromotor response increased significantly 3 days after TNBS intra-rectalcolonic injection in rats. Electroacupuncture on either ST25 or ST36 suppressed the visceromotor response to colorectal distension, but not LI4, at 3 days after TNBS injection. Pretreatment of naltrexone (10 mg/kg, i.p.), opioids antagonist, inhibited the VMR suppress of 10Hz EA to ST36 but not phentolamine (5 mg/kg, i.p.). Pretreatment of either naltrexone or phentolamine inhibited effects of 10Hz EA to ST25. Conclusions: Data show that EA at either ST25 or ST36 potently inhibits hypersensitivity of colorectum after TNBS induced colitis and is differently mediated through the endogenous opioid system and adrenergic system.

• Archives of Pharmacal Research
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• v.29 no.4
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• pp.310-317
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• 2006

We investigated the effects of trinitrobenzene sulfonic acid (TNBS), an amino-group reagent, on the human ether-a-go-go-related gene (HERG) $K^+$ channels expressed in Xenopus oocytes. TNBS neutralizes the positively charged amino-groups of peptide N-terminal and lysine residues. External application of TNBS at 10 mM for 5 min irreversibly shifted the curves for currents at the end of the pulse and tail currents of HERG to a more negative potential and decreased the maximal amplitude of the $I_{tail}$ curve $(I_{tail,max})$. TNBS had little effect on either the activated current-voltage relationship or the reversal potential of HERG current, indicating that TNBS did not change ion selectivity properties. TNBS shifted the time constant curves of both activation and deactivation of the HERG current to a more hyperpolarized potential; TNBS's effect was greater on channel opening than channel closing. External $H^+$ is known to inhibit HERG current by shifting $V_{1/2}$ to the right and decreasing $I_{tail,max}$. TNBS enhanced the blockade of external $H^+$ by exaggerating the effect of $H^+$ on $I_{tail,max}$, not on $V_{1/2}$. Our data provide evidence for the presence of essential amino-groups that are associated with the normal functioning of the HERG channel and evidence that these groups modify the blocking effect of external $H^+$ on the current.

• Fisheries and Aquatic Sciences
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• v.4 no.3
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• pp.144-149
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• 2001

In other to establish the exact determination method of amino nitrogen (AN) in salt-fermented fish sauces, we determined the AN in fish sauces according to the measuring methods and also investigated the main factors influencing on determination method of AN. AN in salt-fermented anchovy sauce increased linearly as fermentation progressed, and was shown the highest amount measuring by the Formol method, followed by the trinitrobenzene sulfonic acid (TNBS) method and the Copper-salt method. AN concentration in anchovy sauces fermented for 12 months was $88.2\%$ and $77.6\%$ for the TNBS method and the Copper-salt method, respectively, on the basis of Formol method. The ratio of AN/total nitrogen (TN) in anchovy sauce fermented for 12 months was higher than that in commercial anchovy sauces. The determination of AN in anchovy sauce by the TNBS method was not affected by salt concentration, and slightly affected by heating. The effect of MSG on AN contents by Copper-salt method was shown higher than those by the Formol method and the TNBS method. The TNBS method was adaptable to measure the content of AN in fish sauce by this study.

• Korean Journal of Acupuncture
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• v.24 no.4
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• pp.131-149
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• 2007

Objectives : The purpose of the present study is to evaluate the effects of herbal acupuncture (HA) with Moxi-tar for the treatment to intestinal disease in mice with 2, 4, 6 - trinitrobenzenesulfonic acid (TNBS) induced colitis. Methods : Mice were administered with 5% TNBS at day 1 and day 7. To investigate effects of HA with Moxi-tar at LI11, treatments were carried out at day -1, day 1, day 3, day 5, and day 7. It was checked on the weight and width of colon, diarrhea, edema, survival rate, changes of body weight, and myeloperoxygenase (MPO) activity. Furthermore, we carried out immunohistochemical staining and Western blot and analyzed mRNA expression by RT-PCR. Results : HA of Moxi-tar at LI11 in preventive mode suppressed macroscopic damages and damages of intestinal epithelial cells and infiltration of immune cells in the colon by TNBS. HA in early and preventive mode ameliorated various symptoms by TNBS. TNBS injection increased MPO activity in colon while HA in preventive mode suppressed increase of MPO activity. HA down-regulated NF-kB activity and reduced expression of TNF-a, IL-1b, and ICAM-1 in colon of TNBS treated mice. Similar to experiment at colon, HA down-regulated NF-kB activity and reduced expression of TNF-a, IL-1b, and ICAM-1 by TNBS in mesenteric lymph node. HA in therapeutic mode suppressed errosion and shortening of colon and MPO activity by TNBS and suppressed mRNA expression of TNF-a, IL-1b, and ICAM-1 in the colon. Conclusions : This study demonstrates that HA with Moxi-tar at LI11 represents a potential therapeutic method of inflammatory bowel diseases.

• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.71-78
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• 2020

Lactobacillus sakei S1 strongly inhibits the expression of interleukin (IL)-6 and IL-1β in lipopolysaccharide-induced peritoneal macrophages by a mechanism for which lactic acid bacteria from kimchi that inhibit tumor necrosis factor-alpha (TNF-α) were isolated. Therefore, we further evaluated the protective effect of this strain on the colitis mouse model induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS). TNBS significantly elevated myeloperoxidase (MPO) expression, macroscopic scores, and colon shortening. Oral L. sakei S1 administration resulted in reduction of TNBS-induced loss in body weight, colon shortening, MPO activity, expression of cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS) and nuclear factor-kappa B (NF-κB). L. sakei S1 inhibited the expression of inflammatory cytokines IL-1β, IL-6 and TNF-α, induced by TNBS, but enhanced IL-10 expression. L. sakei S1 showed resistance to artificial digestive juices and adherence to intestinal epithelial Caco-2 cells. Thus, L. sakei S1 may inhibit the NF-κB pathway and be used in functional food to treat colitis.