• 제목/요약/키워드: single cell injection

검색결과 133건 처리시간 0.026초

Trimethyltin에 의한 랫드 해마의 신경세포 사멸과 iNOS의 연관성 (Inducible nitric oxide synthase is involved in neuronal death induced by trimethyltin in the rat hippocampus)

  • 장석원;최성영;박창남;안미정;신태균;김승준
    • 대한수의학회지
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    • 제51권3호
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    • pp.185-191
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    • 2011
  • Trimethyltin chloride (TMT) has been used as a neurotoxin for inducing brain dysfunction and neuronal death. Neuronal death in the hippocampus by TMT may generate excessive nitric oxide, but there are few studies about nitric oxide synthase enzyme involved in the synthesis of nitric oxide. The purpose of present study is to analyze the TMT toxicity in each region of rat hippocampus. To evaluate the involvement of nitric oxide, we analyzed the effects of aminoguanidine known as a selective inhibitor for inducible nitric oxide synthase on behavioral changes and the hippocampus of rat by TMT toxicity. 6-week-old male Sprague-Dawley rats were administered with a single dose of TMT (8 mg/kg b.w., i.p.) and the control group was similarly administered with distilled water. TMT + aminoguanidine-treated groups were administered with aminoguanidine (10 mg/kg or 100 mg/kg b.w., i.p.) for 3 days prior to TMT injection. The rats were sacrificed 2 days after TMT administration. In the TMT-treated group, a number of cell losses were seen in CA1, CA3 and the dentate gyrus. In the TMT + aminoguanidine-treated group, neuronal death was seen in CA1 and CA3, but reduced in the dentate gyrus compared to the TMT-treated group. Western blot analysis showed that cleaved caspase-3 expression was increased in the TMT-treated group compared to the control group. However, the expression significantly declined in the TMT + aminoguanidine-treated group. The present findings suggest that inducible nitric oxide synthase is involved in neuronal death induced by TMT.

Superovulation and Transplantation of Demi- and Aggregated Embryos in Rabbits

  • Ju, J.C.;Chang, Y.C.;Huang, W.T.;Tang, P.C.;Cheng, S.P.
    • Asian-Australasian Journal of Animal Sciences
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    • 제14권4호
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    • pp.455-461
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    • 2001
  • The effect of exogenous gonadotrophins on superovulation in rabbits was examined. One hundred and sixteen sexually mature California, Chinchilla and New Zealand White rabbits were randomly allocated to control (100 IU hCG), PMSG-treated (100 IU HCG following 150 IU PMSG) and FSH-treated groups (0.3 mg/head /12 h for 3 days followed by 100 IU hCG). All does were mated after hCG injection and were sacrificed or laparotomized within 1 to 4 days postcoitus for counting the number of ovulation points. The number of ovulations was higher in FSH-treated animals than in the control and PMSG-treated groups (37.2 vs. 10.4 and 14.5, p<0.05). Follicle haemorrhagicum was observed in many cases in the PMSG-treated group. No significant difference in ovulation number was observed between left and right ovaries regardless of gonadotropin treatment. In another experiment, 2-cell stage embryos were collected at 26 h postmating and blastomeres were separated by mechanical pipetting or gentle pressure with a fine glass needle. Aggregated or chimeric embryos were produced from two single blastomeres from two breeds, New Zealand White and Chinchlla, with different coat colors. All the embryos were cultured in Ham's F-10 medium supplemented with 1.5% BSA (bovine serum albumin fraction V) and 10% PRS (pregnant rabbit serum), and incubated in a humidified atmosphere with 5% $CO_2$ at $38^{\circ}C$. After development to morula or early blastocyst, the embryos were transferred into the oviducts of recipient does. Results showed that 7 out of 10 does (70%) receiving intact embryos (control) became pregnant and 41 kits were delivered. However, no pregnancy was obtained from the recipient of either denuded demi- or aggregated embryos. It is suggested that embryos without zona pellucida could not develop to term in rabbits.

Rat에서 2, 3, 7, 8-Tetrachlorodibenzo-p-dioxin 유도 독성에 대한 귀뚜라미 추출물의 보호효과 (Preventive Effects of Cricket Extracts, Gryllus bimaculatus, against Toxicity Induced by 2,3,7,8-Tetrachlorodibenzo-p-dioxin in Rats)

  • 이남진;박종배;김동규;윤치영;배형준;조정희;강종구;김윤배;황석연
    • 대한임상검사과학회지
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    • 제36권2호
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    • pp.258-268
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    • 2004
  • This study was carried out to investigate preventive effects of extracts of cricket, Gryllus bimaculatus, against the 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD)-induced toxicity in 7-week-old Sprague-Dawley male rats. Thirty five male rats were divided into 5 groups: one normal control group treated with vehicle and saline (G1); one TCDD-treated group by single intraperitoneal injection (G2); three preventive groups (G3, G4, and G5). The last three groups, G3, G4, and G5, were fed on cricket extracts (50, 100, and 200 mg/kg, respectively) for 2 weeks before TCDD treatment. Various harmful effects were shown by TCDD treatment. The body weights of rats were lost by TCDD. In addition, severe hypertrophy and color change, and the weights gaining were found in the livers of TCDD-treated rats. It was observed that the cytoplasmic vacuolizations and inflammatory cell infiltration around portal triad in the liver. TCDD also elevated the serum activity levels of alanine transaminase(ALT) and aspartate transaminase(AST). However, those losses were compensated by cricket extracts treatment at the level of 200 mg/kg. These findings indicate that cricket extracts may have protective effects against TCDD-induced toxicities in rats.

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익령탕과 익령탕가미방이 Puromycin Aminonucleoside로 유발된 백서의 현증에 미치는 영향 (The Study on the Effect of Ikryung-tang(yiling-tang) and Ikryung-tangkamibang(yiling-tangjiilweifang) on Rats with Nephrosis Induced by Puromycin Aminonucleoside)

  • 조충식;김철중
    • 대한한의학회지
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    • 제23권1호
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    • pp.156-169
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    • 2002
  • Objectives : This experimental study was performed to prove the effect of Ikryung-Tang (yiling-tang; IRT) and Ikryung-tangkamibang (yiling-tangjiaweifang; IRT I, IRT II) on rats with nephrosis induced by a single intravenous injection of puromycin aminonucleoside (PAN). Methods : The effects of IRT, IRT I and IRT II on PAN nephrosis were evaluated by measuring the gene expression of IL-4, $IFN-{\gamma}$ and the concentrations of IgE, albumin, total protein, cholesterol, triglyceride, creatinine, and BUN in the serum and the volume & amount of protein of the 24hrs' urine. Results : In the gene expression linked T cell specialization, IRT II inhibited IL-4 and IgE but IRT and IRT I showed no significant difference compared with control group. On the other hand, IRT, IRT I and IRT II increased $IFN-{\gamma}$ compared with the control group. In the urine protein, serum albumin, total protein, BUN, and creatinine, IRT I especially showed more. significant effect than other groups. In the serum cholesterol and triglyceride, IRT II especially showed more significant effect than other groups. In the urine volume during 24 hrs, IRT especially showed more significant effect than other groups. Conclusions : According to the above results, it is suggested that IRT is effective for the treatment of edema, IRT I is effective for the treatment of hypoproteinemia and kidney dysfunction, IRT II is effective for immune modulation and the treatment of cholesteremia. Therefore IRT, IRT I and IRT II seem to be available for treating nephrosis in clinical practice.

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Pilocarpine투여(投與) 가토폐장(家免肺臟)의 압력(壓力)-용적곡선(容積曲線) 및 폐포표면활성물질(肺胞表面活性物質)에 관(關)하여 (Effects of Pilocarpine on Pressure-Volume Curve of the Excised Rabbit Lung and Pulmonary Surfactant)

  • 이승정;채수희;이석강
    • The Korean Journal of Physiology
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    • 제8권2호
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    • pp.49-53
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    • 1974
  • 제 II 형(第 II 型) 폐포세포(肺胞細胞)의 페포표면활성물질(肺胞表面活性物質) 분비기능(分泌機能)이 부교감신경성작용(副交感神經性作用)에 의(依)해서 영향(影響)을 받을 것인지를 구명(究明)하기 위해서 체중(體重) 매(每) kg 당(當) 0.5mg의 pilocarpine hydrochloride를 투여(投與)한 가토(家兎)에서 폐장(肺臟)의 압력(壓力)-용적곡선(容積曲線)및 폐세척액(肺洗滌液)의 표면활성도(表面活性度)를 측정(測定)한 결과(結果)는 다음과 같다. (1) 적출가토폐(摘出家兎肺)에 공기(空氣)를 주입(注入) 및 배출(排出)할 때의 압력(壓力)-용적곡선(容積曲線)은 정상(正常) 가토(家兎)의 그것보다 hysteresis현상(現象)이 더욱 심(甚)하였으며 특(特)히 배출곡선(排出曲線)이 좌측(左側)으로 편입(偏入)되어있었다. (2) 정상(正常) 가토폐세척액(家兎肺洗滌液)의 평균최대(平均最大) 및 최소표면장력(最小表面張力) 및 stability index는 각각(各各) 46.6 dyne/cm, 7.4 dyne/cm 및 1.5이었다. (3) Pilocarpine 투여가토폐세척액(投與家肺洗滌液)의 평균최대(平均最大) 및 최소표면장력(最小表面張力) 및 stability index는 각각(各各) 47.5 dyne/cm, 6.1 dyne/cm 및 1.5로서 최소표면장력(最小表面張力)은 감소(減少)하는 경향(傾向)을 보였다. (4) 이상의 실험결과(實驗結果)로 미루어 보아 pilocarpine 투여(投與)에 의(依)해서 제 II 형(第 II 型) 폐포세포(肺胞細胞)의 페포표면활성물질(肺胞表面活性物質)의 분비기능(分泌機能)이 증가(增加)되는 경향(傾向)을 보였다.

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실험동물 창상 모델에서 겐타마이신 함유 콜라겐 스폰지의 생체적합성 평가 (Evaluation for Biocompatibility of Gentamicin-collagen Sponge on the Experimental Animal Wound Model)

  • 윤성호;박지희;박정규;김만섭;김달원;송문용;강규일;황원구;구세광;장광호;권영삼
    • 한국임상수의학회지
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    • 제32권5호
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    • pp.404-409
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    • 2015
  • 본 연구의 목적은 겐타마이신 함유 스폰지인 겐타콜과 콜라템프를 랫드 근육내 이식 후 생체적합성을 비교하는 것이다. SD 랫드 66 마리를 네 그룹으로 나누었다; (1) 대퇴사두근에 무처치한 군 (대조군, 6 마리), (2) 대퇴사두근에 겐타마이신 액을 주사한 군 (겐타마이신 군, 6 마리), (3) 대퇴사두근에 콜라템프를 이식한 군 (콜라템프 군, 27마리), (4) 대퇴사두근에 겐타콜을 이식한 군 (겐타콜 군, 27 마리). 겐타콜과 콜라템프 이식 한근육 내 겐타마이신 농도는 시간이 지남에 따라 점차 감소하였다. 혈액 내에 겐타마이신 농도는 측정되지 않았다. 조직학적으로 겐타마이신액을 주사 후 근육 내에 다형핵백혈구, 림프구, 대식세포를 포함하는 염증세포가 중등도에서 심하게 침윤되었고, 경도에서 중등도의 근육내 부종이 관찰되었다. 그러나, 이러한 국소 자극의 조직학적 변화는 콜라템프와 겐타콜 군에서는 현저히 감소하였다. 이상의 결과, 겐타콜은 콜라템프와 비교하였을 때에 전신적인 영향을 미치지 않으며 국소자극의 정도가 유사하였고, 생체이용성이 유사하여 좋은 생체적합성을 가지는 것으로 생각된다.

족삼리(足三里) 사과락약침(絲瓜絡藥鍼)이 생쥐의 Collagen-induccd Arthritis에 미치는 영향 (A Study on the Effect of Luffae Fructus Retinervus Herbal-Acupuncture at ST36 on Collagen-induced Arthritis in Mice)

  • 채충헌;최선미;임윤경
    • Journal of Acupuncture Research
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    • 제22권1호
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    • pp.131-144
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    • 2005
  • 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)이 DBA/lJ mouse의 collagen-induced arthritis 미치는 영향(影響)을 관찰(觀察) 한 결과(結果), 다음과 같은 결론(結論)을 얻었다. 1. 세포독성(細胞毒性) 측정결과(測定結果), 사과락약침액(絲瓜絡藥鍼液) 1%의 농도(濃度)에서 가장 높은 세포생존률(細胞生存率)이 관찰(觀察)되었다. 2. In vitro screening 결과(結果), 사과락약광액(絲瓜絡藥鑛液) 1%의 농도(濃度)에서 CIA mouse 관절활막세포(關節滑膜細胞)의 TNF-${\alpha}$발현(發現)이 유의(有意)하게 감소(減少)하였다. 3. 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 관절염(關節炎) 발병률(發病率)이 감소(減少)하였다. 4. 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 CIA mouse의 각장(脚臟)무게가 감소(減少)하였다. 5. 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 CIA mouse의 혈청(血淸) IL-6, INF-${\gamma}$, TNF-${\alpha}$, IgG, IgM 및 Anti-collagen Ⅱ가 유의(有意)하게 감소(減少)하였다. 6. 조직학적(組織學的) 검사(檢査) 결과(結果), 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 CIA mouse 관절(關節)의 연골파괴(軟骨破壞)와 활막증식(滑膜增殖)이 감소(減少)되고 교원질섬유(膠原質纖維)가 정상군(正常群)과 유사(類似)하게 유지(維持)되었다. 7. 임파절내(淋巴節內) CD3e+와 CD19+세포(細胞) 비율(比率), CD4+와 CD8+세포(細胞) 비율(比率)은 사과락약광치료군(絲瓜絡藥鑛治療群)에서 정상군(正常群)과 유사(類似)하게 유지(維持)되었다. 8. 임파절내(淋巴節內) CD69+/CD3e+세포(細胞) 및 CD11a+/CD19+세포(細胞)와 관절내(關節內) CD11b+/Gr-1+세포(細胞)는 사과락약침치료군(絲瓜絡藥鍼治療群)에서 대조군(對照群)에 비하여 유의(有意) 하게 감소(減少)하였다.

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BCG가 Ehrlich 암세포를 이식한 생쥐의 위점막 상피세포의 DNA합성 및 미세구조에 미치는 영향 (Effects of BCG on the DNA Synthesis and Ultrastructure of Mouse Gastric Mucosal Epithelial Cells Inoculated with Ehrlich Carcinoma Cells)

  • 고정식;류인상;박경호;박대균
    • Applied Microscopy
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    • 제39권3호
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    • pp.205-218
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    • 2009
  • 이 실험은 Ehrlich 종양세포를 이식한 후 BCG를 투여하였을 때, 위점막 상피세포의 형태학적 변화와 DNA합성능의 변화를 연구하고자 시행하였다. 실험동물로는 체중 25 g 내외의 성숙한 생쥐(ICR계통)를 정상대조군, 종양세포이식대조군(종양대조군), 종양세포이식후 BCG투여군(BCG투여군)으로 구분하였다. 종양대조군과 BCG투여군 동물들은 샅부위 피하에 각각 $1{\times}10^7$의 Ehrlich 종양세포를 이식한 후, 다음날부터 BCG ($0.6{\times}10^8{\sim}6.4{\times}10^8$ CFU, 27 mg/vial, Connaught Lab., Canada)를 하루건너 한 번씩 피부밑조직에 주사하였으며, 종양대조군은 종양세포이식 후에 BCG 대신 0.2mL의 생리식염수를 피부밑조직에 주사하였다. 자기방사법적 관찰을 위해서는 BCG를 마지막으로 주사한 다음날 $^3H$-thymidine (methyl-$^3H$-thymidine: specific activity 25 Ci/mmol, Amersham Lab., England) 0.7${\mu}Ci/gm$를 꼬리정맥에 주사하고, 70분 후 도살하여 위조직을 떼어내어 10% formalin에 고정하였다. 자기방사표본관찰은 위점막 조직이 세로로 잘 절단된 부위를 택하여 점막근육판을 따라 점막길이 3.5mm의 위점막 조직에 분포하는 $^3H$-thymidine 표지세포의 수를 계수하였으며, 일반조직 관찰을 위해서는 hematoxylin-eosin (H-E)염색을 시행하였다. 전자현미경 관찰을 위해서는 떼어낸 위조직을 2.5% glutaraldehyde-1.5% paraformaldehyde 혼합액에 고정한 후, 1% osmium tetroxide 액에 다시 고정하였으며, 고정이 끝난 조직은 탈수과정을 거쳐 araldite 혼합액에 포매하였다. 광학현미경적 관찰에서 종양대조군과 BCG투여군은 위점막 조직에서 형태적으로 큰 변화를 볼 수 없었다. 전자현미경적 관찰에서 BCG투여군의 점액상피세포는 전체적인 모습이 정상대조군과 종양대조군의 소견과 유사하였으나 정상대조군과 종양대조군에 비해 수초구조와 뭇소포체(multivesicular body) 및 전자밀도가 높은 큰 미토콘드리아속과립이 자주 관찰되었다. 자기방사법적 관찰에서 정상대조군, 종양대조군, BCG투여군은 점막길이 3.5 mm 당 출현하는 표지세포수가 각각 380.2 (${\pm}31.35$), 426.1 (${\pm}28.43$) 및 301.8 (${\pm}34.63$)개이었으며, BCG투여군은 정상대조군과 종양대조군에 비하여 표지된 은입자의 수가 매우 적어서 표지과립이 겨우 구별될 정도의 세포가 많이 관찰되었다. 이상의 결과를 종합해보면 Ehrlich 종양세포를 이식한 동물에 BCG를 반복 투여하면 위점막 상피세포의 DNA합성을 효과적으로 억제하면서도 형태적인 변화가 매우 경미하였으며, 이러한 결과는 BCG가 항암치료 시 보조제로 사용할 수 있는 좋은 약제라고 생각된다.

흡수성 차폐막에 접목된 두개관골세포의 골조직 재생에 미치는 영향 (Effect of Calvarial Cell Inoculated Onto the Biodegradable Barrier Membrane on the Bone Regeneration)

  • 유부영;이만섭;권영혁;박준봉;허익
    • Journal of Periodontal and Implant Science
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    • 제29권3호
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    • pp.483-509
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    • 1999
  • Biodegradable barrier membrane has been demonstrated to have guided bone regeneration capacity on the animal study. The purpose of this study is to evaluate the effects of cultured calvarial cell inoculated on the biodegradable barrier membrane for the regeneration of the artificial bone defect. In this experiment 35 Sprague-Dawley male rats(mean BW 150gm) were used. 30 rats were divided into 3 groups. In group I, defects were covered periosteum without membrane. In group II, defects were repaired using biodegradable barrier membrane. In group III, the defects were repaired using biodegradable barrier membrane seeded with cultured calvarial cell. Every surgical procedure were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). After anesthesia, 5 rats were sacrificed by decapitation to obtain the calvaria for bone cell culture. Calvarial cells were cultured with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. The number of cell inoculated on the membrane were $1{\times}10^6$ Cells/ml. The membrane were inserted on the artificial bone defect after 3 days of culture. A single 3-mm diameter full-thickness artificial calvarial defect was made in each animal by using with bone trephine drill. After the every surgical intervention of animal, all of the animals were sacrificed at 1, 2, 3 weeks after surgery by using of perfusion technique. For obtaining histological section, tissues were fixed in 2.5% Glutaraldehyde (0.1M cacodylate buffer, pH 7.2) and Karnovsky's fixative solution, and decalcified with 0.1M disodium ethylene diaminetetraacetate for 3 weeks. Tissue embeding was performed in paraffin and cut parallel to the surface of calvaria. Section in 7${\mu}m$ thickness of tissue was done and stained with Hematoxylin-Eosin. All the specimens were observed under the light microscopy. The following results were obtained. 1 . During the whole period of experiment, fibrous connective tissue was revealed at 1week after surgery which meant rapid soft tissue recovery. The healing rate of defected area into new bone formation of the test group was observed more rapid tendency than other two groups. 2 . The sequence of healing rate of bone defected area was as follows ; test group, positive control, negative control group. 3 . During the experiment, an osteoclastic cell around preexisted bone was not found. New bone formation was originated from the periphery of the remaing bone wall, and gradually extended into central portion of the bone defect. 4 . The biodegradable barrier membrane was observed favorable biocompatibility during this experimental period without any other noticeable foreign body reaction. And mineralization in the newly formed osteoid tissue revealed relatively more rapid than other group since early stage of the healing process. Conclusively, the cultured bone cell inoculated onto the biodegradable barrier membrane may have an important role of regeneration of artificial bone defects of alveolar bone. This study thus demonstrates a tissue-engineering the approach to the repair of bone defects, which may have clinical applications in clinical fields of the dentistry including periodontics.

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Oxytocin produces thermal analgesia via vasopressin-1a receptor by modulating TRPV1 and potassium conductance in the dorsal root ganglion neurons

  • Han, Rafael Taeho;Kim, Han-Byul;Kim, Young-Beom;Choi, Kyungmin;Park, Gi Yeon;Lee, Pa Reum;Lee, JaeHee;Kim, Hye young;Park, Chul-Kyu;Kang, Youngnam;Oh, Seog Bae;Na, Heung Sik
    • The Korean Journal of Physiology and Pharmacology
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    • 제22권2호
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    • pp.173-182
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    • 2018
  • Recent studies have provided several lines of evidence that peripheral administration of oxytocin induces analgesia in human and rodents. However, the exact underlying mechanism of analgesia still remains elusive. In the present study, we aimed to identify which receptor could mediate the analgesic effect of intraperitoneal injection of oxytocin and its cellular mechanisms in thermal pain behavior. We found that oxytocin-induced analgesia could be reversed by $d(CH_2)_5[Tyr(Me)^2,Dab^5]$ AVP, a vasopressin-1a (V1a) receptor antagonist, but not by $desGly-NH_2-d(CH_2)_5[D-Tyr^2,Thr^4]OVT$, an oxytocin receptor antagonist. Single cell RT-PCR analysis revealed that V1a receptor, compared to oxytocin, vasopressin-1b and vasopressin-2 receptors, was more profoundly expressed in dorsal root ganglion (DRG) neurons and the expression of V1a receptor was predominant in transient receptor potential vanilloid 1 (TRPV1)-expressing DRG neurons. Fura-2 based calcium imaging experiments showed that capsaicin-induced calcium transient was significantly inhibited by oxytocin and that such inhibition was reversed by V1a receptor antagonist. Additionally, whole cell patch clamp recording demonstrated that oxytocin significantly increased potassium conductance via V1a receptor in DRG neurons. Taken together, our findings suggest that analgesic effects produced by peripheral administration of oxytocin were attributable to the activation of V1a receptor, resulting in reduction of TRPV1 activity and enhancement of potassium conductance in DRG neurons.