Kinetics of IFN-Gamma and TNF-Alpha Gene Expression and Their Relationship with Disease Progression after Infection with Mycobacterium Tuberculosis in Guinea Pigs

노인순,조성애,엄석용,조상래 연세대학교의과대학 2013 Yonsei medical journal Vol.54 No.3

Purpose: Guinea pig is one of the most suitable animal models for Mycobacterium tuberculosis (M. tb) infection since it shows similarities to pulmonary infection in humans. Although guinea pig shows hematogenous spread of M. tb infection into the whole body, immunological studies have mainly focused on granulomatous tissues in lungs and spleens. In order to investigate the time-course of disease pathogenesis and immunological profiles in each infected organ, we performed the following approaches with guinea pigs experimentally infected with M. tb over a 22-week post-infection period. Materials and Methods: We examined body weight changes, M. tb growth curve, cytokine gene expression (IFN-γ and TNF-α), and histopathology in liver, spleen, lungs and lymph nodes of infected guinea pigs. Results:The body weights of infected guinea pigs did not increase as much as uninfected ones and the number of M. tb bacilli in their organs increased except bronchotracheal lymph node during the experimental period. The gene expression of IFN-γ and TNF-α was induced between 3 and 6 weeks of infection; however, kinetic profiles of cytokine gene expression showed heterogeneity among organs over the study period. Histophathologically granulomatous lesions were developed in all four organs of infected guinea pigs. Conclusion: Although IFN-γ and TNF-α gene expression profiles showed heterogeneity, the granuloma formation was clearly observed in every organ regardless of whether the number of bacilli increased or decreased. However, this protective immunity was accompanied with severe tissue damage in all four organs, which may lead to the death of guinea pigs.

Phospholipase D in Guinea Pig Lung Tissue Membrane is Regulated by Cytosolic ARF Proteins

( Yean Jun Chung ),( Jin Rak Jeong ),( Byung Chul Lee ),( Ji Young Kim ),( Young In Park ),( Jai Youl Ro ) 한국미생물생명공학회 2003 Journal of microbiology and biotechnology Vol.13 No.6

The Effect of Trimebutine on the Overlap Syndrome Model of Guinea Pigs

( Zahid Hussain ),( Da Hyun Jung ),( Young Ju Lee ),( Hyojin Park ) 대한소화기기능성질환·운동학회(구 대한소화관운동학회) 2018 Journal of Neurogastroenterology and Motility (JNM Vol.24 No.4

Background/Aims Functional dyspepsia (FD) and irritable bowel syndrome (IBS) are common gastrointestinal (GI) disorders and these patients frequently overlap. Trimebutine has been known to be effective in controlling FD co-existing diarrhea-dominant IBS, however its effect on overlap syndrome (OS) patients has not been reported. Therefore, we investigated the effect of trimebutine on the model of OS in guinea pigs. Methods Male guinea pigs were used to evaluate the effects of trimebutine in corticotropin-releasing factor (CRF) induced OS model. Different doses (3, 10, and 30 mg/kg) of trimebutine were administered orally and incubated for 1 hour. The next treatment of 10 μg/kg of CRF was intraperitoneally injected and stabilized for 30 minutes. Subsequently, intragastric 3 mL charcoal mix was administered, incubated for 10 minutes and the upper GI transit analyzed. Colonic transits were assessed after the same order and concentrations of trimebutine and CRF treatment by fecal pellet output assay. Results Different concentrations (1, 3, and 10 μg/kg) of rat/human CRF peptides was tested to establish the OS model in guinea pigs. CRF 10 μg/kg was the most effective dose in the experimental OS model of guinea pigs. Trimebutine (3, 10, and 30 mg/kg) treatment significantly reversed the upper and lower GI transit of CRF induced OS model. Trimebutine significantly increased upper GI transit while it reduced fecal pellet output in the CRF induced OS model. Conclusions Trimebutine has been demonstrated to be effective on both upper and lower GI motor function in peripheral CRF induced OS model. Therefore, trimebutine might be an effective drug for the treatment of OS between FD and IBS patients. (J Neurogastroenterol Motil 2018;24:669-675)

Cholesterol-induced inflammation and macrophage accumulation in adipose tissue is reduced by a low carbohydrate diet in guinea pigs

David Aguilar,Ryan C deOgburn,Jeff S Volek,Maria Luz Fernandez 한국영양학회 2014 Nutrition Research and Practice Vol.8 No.6

BACKGROUND/OBJECTIVES: The main objective of this study was to evaluate the effects of a high cholesterol (HC) dietary challenge on cholesterol tissue accumulation, inflammation, adipocyte differentiation, and macrophage infiltration in guinea pigs. A second objective was to assess whether macronutrient manipulation would reverse these metabolic alterations. MATERIALS/METHODS: Male Hartley guinea pigs (10/group) were assigned to either low cholesterol (LC) (0.04g/100g) or high cholesterol (HC) (0.25g/100g) diets for six weeks. For the second experiment, 20 guinea pigs were fed the HC diet for six weeks and then assigned to either a low carbohydrate (CHO) diet (L-CHO) (10% energy from CHO) or a high CHO diet (H-CHO) (54% CHO) for an additional six weeks. RESULTS: Higher concentrations of total (P < 0.005) and free (P < 0.05) cholesterol were observed in both adipose tissue and aortas of guinea pigs fed the HC compared to those in the LC group. In addition, higher concentrations of pro-inflammatory cytokines in the adipose tissue (P < 0.005) and lower concentrations of anti-inflammatory interleukin (IL)-10 were observed in the HC group (P < 0.05) compared to the LC group. Of particular interest, adipocytes in the HC group were smaller in size (P < 0.05) and showed increased macrophage infiltration compared to the LC group. When compared to the H-CHO group, lower concentrations of cholesterol in both adipose and aortas as well as lower concentrations of inflammatory cytokines in adipose tissue were observed in the L-CHO group (P < 0.05). In addition, guinea pigs fed the L-CHO exhibited larger adipose cells and lower macrophage infiltration compared to the H-CHO group. CONCLUSIONS: The results of this study strongly suggest that HC induces metabolic dysregulation associated with inflammation in adipose tissue and that L-CHO is more effective than H-CHO in attenuating these detrimental effects.

The Effects of an Extract of Atractylodes Japonica Rhizome, SKI3246 on Gastrointestinal Motility in Guinea Pigs

( Jae Jun Park ),( Nu Ri Chon ),( Young Ju Lee ),( Hyojin Park ) 대한소화기기능성질환·운동학회 2015 Journal of Neurogastroenterology and Motility (JNM Vol.21 No.3

Background/Aims: There are limited therapeutic options available for irritable bowel syndrome with diarrhea (IBS-D). We tested the effects of Atractylodes japonica rhizome, a perennial plant native to North Asia, on both upper and lower gastrointestinal (GI) motility in guinea pigs. Methods: The extract of A. japonica rhizome was administered orally at different doses to test its effects on upper GI motility as determined from charcoal transit in native guinea pigs and in guinea pigs pretreated with thyrotropin-releasing hormone or mustard oil. Regarding its effect on lower GI motility, the removed guinea pig colon was suspended in a chamber containing Krebs-Henseleit solution and the transit time of artificial feces was measured with various dilutions of the extract. As for in vivo assay, weight and number of fecal pellets expelled were determined under the same drug preparation used in upper GI motility experiment. Results: The extract of A. japonica rhizome had no significant effect on upper GI motility in either normal or altered physiological states. However, the extract increased colonic transit time in the in vitro model. In the fecal expulsion study, the cumulative weight and number of pellets did not differ significantly between the control group and groups treated with the extracts. In the animals pretreated in vivo with thyrotropin-releasing hormone, however, the weight and number of fecal pellets were significantly decreased in animals treated with 300 mg/kg and 600 mg/kg doses of extract. Conclusions: Our findings suggest that the extract of A. japonica rhizome can be a potential agent for IBS-D. (J Neurogastroenterol Motil 2015;21:352-360)

Immunization of guinea pigs with <i>Salmonella</i> delivered anti-<i>Brucella</i> formulation reduces organs bacterial load and mitigates histopathological consequences of <i>Brucella abortus</i> 544 challenge

Lalsiamthara, Jonathan,Lee, John Hwa Elsevier 2018 Veterinary immunology and immunopathology Vol.195 No.-

<P><B>Abstract</B></P> <P>With an objective to generate safe and effective anti-<I>Brucella</I> vaccine, an attenuated live <I>Salmonella</I> Typhimurium vector delivering heterologous <I>Brucella</I> immunogenic proteins SOD, Omp19, BLS, and PrpA formulated with purified <I>Brucella abortus</I> lipopolysaccharide was evaluated on a guinea pig model. This model represents high susceptibility to <I>Brucella</I> infections and showed similarities in reproducing human pathologies. On safety perspectives, the vaccine formulation induced no observable alterations on general health and histology of the vaccinated guinea pigs. Upon virulent strain 544 challenge, a protective index of 1.52 was observed based on differential splenic counts. Post-challenge histopathology revealed that <I>Brucella</I> induced microgranulomas and fatty degenerations were prominent in the organs of non-immunized animals as compared to immunized animals. With these findings, it is suggestive that this live <I>Brucella</I>-free vaccine formulation is safe and protective on a sensitive guinea pig model and may be suitable for further human-related vaccine trials.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Assessment of <I>Salmonella</I> vectored anti-<I>Brucella</I> vaccine formulated with lipopolysaccharide in guinea pigs. </LI> <LI> The formulation exhibited safety on general health, postmortem, and histological standpoints. </LI> <LI> The formulation reduces organ challenge bacterial loads and mitigations of histopathological consequences. </LI> </UL> </P>

기니 피그에서 콜레시스토키닌 투여가 고탄수화물 , 저지방 , 저단백질식에 의한 색소성 담석의 형성에 미치는 영향

박용현(Yong Hyun Park),서경석(Kyung Suk Suh),김선희(Sun Whe Kim) 대한소화기학회 1994 대한소화기학회지 Vol.26 No.1

N/A W e reported an animal model of pigment gallstone induced by carbohydrate-rich diet main- ly composed with rice. However, the machanisms have not been clearly understood. Under the hypothesis that high-carbohydrate diet (CHO) might induce gallstone formation via the mechanism of the relative bile stasis caused by low secretion of cholecystokin (CCK), the aim of this study was to examine whether exogenous CCK administration inhibits gall- stone formation induced by CHO in guinea pig. Male guinea pigs were divided into 3 groups(Group 1: control chow-fed group, Group 2: high carbohydrate fed group, Group 3: high cabohydrate fed and CCK injected group). High carbo- hydrate diet was 63.2% carbohydrate(45.8% in control chow), mainly composed of rice, and Group 3 received a daily injection of cholecystokinin(0.5nmol/kg). After 6 weeks of feeding, the guinea pigs were sacrificed. The stones were analyzed by infrared spectrophotornetry and gallbladder bile was analyzed by using commercial kits. In group 1, gallstone was found in one case out of 10 animals, in Group 2, 9 out of 14(p<0.05 vs Group 1) and in Group 3, 3 out of 12(p<0 05 vs Group 2). There were no differences in the concentrations of total calcium, total bilirubin, cholesterol, phospholipid and bile acid of gall- bladder bile among 3 groups. The stones were mainly composed of calcium bilirubinate, cho- lesterol, calcium phosphate, and calcium palmitate, which were similar to human calcium bilirubinate stone. The prevalence of stone formation was lowered with administration of CCK in CHO fed animals. A possible mechanism is that exogenous CCK may recover the low CCK release by CHO. It is suggested that bile stasis caused by poor CCK release which may be due to CHO be the one of the contributing factors in the formation of pigment stones in guinea pigs. (Korean J Gastroenterol 1 994; 26: 151 156)

Inhibitory Effects of Resveratrol on Melanin Synthesis in Ultraviolet B-Induced Pigmentation in Guinea Pig Skin

( Taek Hwan Lee ),( Jae Ok Seo ),( So Hyeon Baek ),( Sun Yeou Kim ) 한국응용약물학회 2014 Biomolecules & Therapeutics(구 응용약물학회지) Vol.22 No.1

Resveratrol is a polyphenolic compound found in various natural products such as grapes and berries and possesses anti-cancer, anti-hyperlipidemia, and anti-aging properties. Recently, it has been reported that resveratrol inhibits a-melanocyte-stimulating hormone signaling, viability, and migration in melanoma cells. However, these effects have not been confirmed in vivo, specifically brownish guinea pigs. To evaluate the potential of resveratrol as a regulator of melanin for hyperpigmentation therapy, the influence of resveratrol on pigmentation was investigated by ultraviolet B-induced hyperpigmentation in brownish guinea pig skin. We found that resveratrol reduced the expression of melanogenesis-related proteins tyrosinase, tyrosinase-related proteins 1 and 2, and microphthalmia-associated transcription factor in melanoma cells. Furthermore, topical application of resveratrol was demonstrated to significantly decrease hyperpigmentation on ultraviolet B-stimulated guinea pig skin in vivo. Based on our histological data, resveratrol inhibits melanin synthesis via a reduction in tyrosinase-related protein 2 among the melanogenic enzymes. This study is the first to provide evidence supporting resveratrol as a depigmentation agent, along with further clinical investigation of resveratrol in ultraviolet B-induced skin disorders such as hyperpigmentation and skin photoaging.

Sol-M®의 피부 자극성 및 감작성 연구

이혜숙,황석연 한국피부과학연구원 2010 대한피부미용학회지 Vol.8 No.1

The use of atopic agents has been recently increased in various kinds of products, although there were some reports that atopic agents might cause allergic contact dermatitis. This study was conducted to investigate the skin irritation and skin sensitization induced by Sol-M® which contains Angelicae Dahuricae Radix, Phellodendri Cortex, Pinus densiflora Sieb., Mori Cortex Radicis in rabbits and guinea pigs according to Korea Food and Drug Agency(KFDA) guidelines for toxicological test. GPMT(The Guinea Pig Maximization Test) method was applied to evaluate the sensitization potential of Sol-M® in Hartley guinea pigs. In primary skin irritation test, rabbits were treated with 5 %, 10 % and 20 % of Sol-M® for 24 hrs. Sol-M® did not induced any adverse reaction such as erythema and edema on intact skin sites, but on abraded skin sites, some rabbits showed edema 24 hrs after topical application. So 5 %, 10 % and 20% of Sol-M® was classified as a practically non-irritating material based on the score 0.21, 0.33 and 0.27 of primary irritation index. In the sensitization test, guinea pigs were sensitized with intradermal injection of 0.1 ml Sol-M® for 24 hrs. After 1 week, Sol-M® was treated on the site of injection and challenged 2 weeks later. 10 % and 20 % of Sol-M® was classified to grade Ⅲ and Ⅴ, indicating moderate and extreme sensitizers, respectively. From results of the present study, it is suggested that 10 % Sol-M® is a practically-safe material to skin without potential of irritation and sensitization. 본 연구는 아토피 피부에 사용되는 천연추출 복합물인 Sol-M®을 면역독성 평가의 일환으로 접촉성 과민반응 유발의 가능성을 접촉성 알러지 시험에 많이 사용되는 동물인 기니픽을 실험동물로 하는 GPMT(The Guinea Pig Maximization Test)를 이용하여 감작 및 피부자극성을 평가하였다. 피부자극시험에서는 Sol-M® 5, 10 그리고 20 % 농도별로 토끼의 피부에 적용한 후 국소적으로 나타나는 피부자극성을 평가하기 위하여 홍반, 가피 및 부종의 형성정도를 관찰하였다. 그 결과 Sol-M® 5, 10 그리고 20 % 처치군에서 제모 한 정상 피부 및 인위적으로 손상을 준 피부에서 홍반과 가피가 형성되지 않았지만 약간의 부종은 확인되었다. 피부자극 평가표 및 일차피부자극지수를 산출한 결과 Sol-M® 5, 10, 20 %의 P.I.I.는 매우 낮은 수준인 0.21, 0.33, 0.27로 확인되어 최고 20 % 농도까지 비자극 물질인 것으로 사료된다. 피부감작시험에서는 반응유도 후 시험물질을 제거하고 최종 관찰한 결과, 용매인 증류수 투여군과 고농도인 Sol-M 10 % 처치군에서 약 50 %의 경미한 피부감작성이 관찰되었다. Sol-M 20 % 처치군은 자극 지수 1의 경도 반응을 나타낸 동물이 6마리 중 5마리가 나타나 등급 Ⅴ(매우강함)로 평가되었다. 이상의 결과를 종합해 볼 때 10 % 이하의 Sol-M®을 임상에 적용할시 약물에 의한 자극성 및 면역반응에 의한 감작성 부작용을 유발하지 않을 것으로 판단된다.

The Effect of Peripheral CRF Peptide and Water Avoidance Stress on Colonic and Gastric Transit in Guinea Pigs

Zahid Hussain,박효진,김혜원,허철웅,이영주 연세대학교의과대학 2017 Yonsei medical journal Vol.58 No.4

Functional dyspepsia (FD) and irritable bowel syndrome (IBS) are common gastrointestinal (GI) diseases; however, there is frequent overlap between FD and IBS patients. Emerging evidence links the activation of corticotropin releasing factor (CRF) receptors with stress-related alterations of gastric and colonic motor function. Therefore, we investigated the effect of peripheral CRF peptide and water avoidance stress (WAS) on upper and lower GI transit in guinea pigs. Dosages 1, 3, and 10 μg/kg of CRF were injected intraperitoneally(IP) in fasted guinea pigs 30 minutes prior to the intragastric administration of charcoal mix to measure upper GI transit. Colonic transits in non-fasted guinea pigs were assessed by fecal pellet output assay after above IP CRF doses. Blockade of CRF receptorsby Astressin, and its effect on GI transit was also analyzed. Guinea pigs were subjected to WAS to measure gastrocolonic transit in different sets of experiments. Dose 10 μg/kg of CRF significantly inhibited upper GI transit. In contrast, there was dose dependentacceleration of the colonic transit. Remarkably, pretreatment of astressin significantly reverses the effect of CRF peptide on GI transit. WAS significantly increase colonic transit, but failed to accelerate upper GI transit. Peripheral CRF peptide significantly suppressed upper GI transit and accelerated colon transit, while central CRF involved WAS stimulated only colonic transit. Therefore,peripheral CRF could be utilized to establish the animal model of overlap syndrome.