Effects of imipramine and lithium on blood alcohol level in rabbits

의학과/석사[한글] [영문] Imipramine, a tricyclic antidepressant, was synthesized by Hafliger in 1948. Kuhn, in 1958, found imipramine was remarkably effective in certain depressed states, especially in endogenous depression. since then, further evidences for the effectiveness of this compound have been accumulated. Lithium, a monovalent cation and the lightest alkali metal, was discovered in 1818 by Arfwedson. Since Cade, in 1949, found that lithium carbonate caused sedation in guinea pigs and that it did calm manic patients, numerous studies have indicated that the drug is effective in the control of mania and other psychotic excitement and also in prevention of manic depressive episodes. It has lately been reported that lithium and several other psychotropic drugs elevated the blood alcohol level in rabbits. In views of these reports the author conducted an animal experiment to investigate the effects of imipramine and lithium, alone or in combination, on blood alcohol level in rabbits. Materials and Methods 1. The experimental work was done on mature rabbits of both sexes, weighing between 2.0 Kg and 3.0 Kg. 2. The experimental animals were divided into 2 groups; the control and the experimental group. 3. The control group was given alcohol alone. 4. The experimental group was divided into 5 groups; Ⅰ) alcohol + lithium, Ⅱ) alcohol + imipramine (I.M.), Ⅲ) alcohol + lithium + imipramine (I.M.), Ⅳ) alcohol + imipramine(oral), and Ⅴ) alcohol + lithium + imipramine(oral) group. 5. Alcohol + imipramine(I.M.) group was further divided into 4 subgroups, according to the time of alcohol injection; alcohol administration immediately, 10, 20, and 30 minutes after imipramine injection. 6. Alcohol + imipramine(oral) group and alcohol + lithium + imipramine(oral) group were further divided into 2 subgroups in which one subgroup was given imipramine for 7 days and another for 14 days. 7. Lithium chloride solution (6.36%) was given in a dose of 3.0 mEq/Kg of body weight daily for 4 days by intravenous route. The last dose was given 1 hour before alcohol administration. 8. Imipramine was given intramuscularly in a single dose of 2.0 mg/Kg of body weight. In case of oral administration imipramine was given in a dose of 4.0 mg/Kg of body weight daily for 7 days or for 14 days. The last dose was given one hour before alcohol administration. 9. In all groups 20% ethanol solution was given in a dose of 5.0 ml/Kg of body weight in 5 minutes by intravenous route. 10. All of the blood specimens were obtained by cardiac puncture at 10 and 30 minutes after alcohol administration. 11. The blood alcohol level determination was made by Cavett's method. Results 1. Alcohol + lithium group Lithium elevated the blood alcohol level significantly both at 10 and 30 minutes after alcohol administration (P〈0.05). 2. Alcohol + imipramine (I.M.) group Imipramine elevated the blood alcohol level significantly both at 10 and 30 minutes after alcohol administration in the first subgroup in which alcohol was given immediately following imipramine injection (P〈0.05). In the second and third subgroup, the blood alcohol level was elevated significantly only at 10 minutes, but not at 30 minutes after alcohol administration. There was no significant change in the blood alcohol level in the fourth subgroup. 3. Alcohol + lithium + imipramine (I.M.) group Imipramine combined with lithium elevated the blood alcohol level significantly both at 10 and 30 minutes after alcohol administration (P〈0.05). 4. Alcohol + imipramine(oral) group Imipramine elevated the blood alcohol level significantly at 10 and 30 minutes after alcohol administration in the subgroups in which imipramine was administered either for 7 or 14 days (P〈0.05). But there was no significant difference in the blood alcohol level between these two subgroups (P〉0.05). 5. Alcohol + lithium + imipramine(oral) group Imipramine(oral) combined with lithium elevated the blood alcohol level significantly at 10 and 30 minutes after alcohol administration in both subgroups in which imipramine was administered either for 7 or 14 days (P〈0.05). The blood alcohol level of these two subgroups was significantly higher than either that of lithium alone group or imipramine(oral) alone group (P〈0.05). 6. Above experimental results were analysed statistically between the control group and each experimental group, and also among the experimental group. Conclusion 1. The intravenous injection of 6.36% lithium chloride solution in a dose of 3.0 mEq/Kg of body weight daily for 4 days elevated the blood alcohol level in rabbits significantly both at 10 and 30 minutes after alcohol administration. 2. Imipramine administered intramuscularly in a single dose of 2.0 mg/kg of body weight showed variable effects on the blood alcohol level according to the time of alcohol administration after imipramine injection. The blood alcohol level was elevated both at 10 and 30 minutes after alcohol administration in the subgroup in which alcohol was administered immediately after imipramine injection. And in the subgroups in which alcohol was administered at 10 and 20 minutes after imipramine injection, the blood alcohol level was elevated only at 10 minutes after alcohol administration. 3. Imipramine administered orally in a dose of 4.0 mg/kg of body weight daily for 7 or 14 days elevated the blood alcohol level significantly both at 10 and 30 minutes after alcohol administration. There was no significant difference in the blood alcohol level between the two subgroups in which imipramine was administered for 7 or 14 days. 4. Imipramine when combined with lithium elevated the blood alcohol level significantly at 10 and 30 minutes after alcohol administration in both orally and intramuscularly administered groups. In the former group the blood alcohol level was significantly higher than that of imipramine alone or lithium alone group.restrictio

Morphological changes of liver by administration of flubendazole to experimental animals

의학과/박사[한글] 구충제로 널리 쓰이고 있는 Mebendazole과 같은 Benzimidazole 유도체인 Flubendazole(Methyl N-[5-(6)-(p-fluorobenzoyl)-2-benzimidazolyl] carbamate, Janssen Pharmaceutica)는 무미, 무취의 미세한 분말약재로 각종 실험동물에서 광범위한 구충효과가 있고, 장 에서 홉수되어 가토의 간흡충증에는 소량의 단회 투여로 완전 구충되었음이 보고되었다.(Sob et al, 1977) 이 약제는 간흡충 충체의 표피층에 있는 protoplasmic tubule에 작용하여 표피층의 대사를 저해함으로써 살충효과의 일부를 담당할 것으로 추정되고 있으나, 아직까지도 본 약제의 약리학적 특성이나 실험동물 또는 인체에 대한 독성은 보고된 바 없다. 또한 Flubendazole은 가토에 감염된 간흡충의 구충력은 강력하나 백서의 간흡충증에는 구충효과가 극소하다고 하였으며, 본 약제가 동물 종(種)간에 상이하게 작용하는 것으로 보고되었다. 그러나 본 약제가 서로 다른 동물종에 투여되었을 때 숙주의 간에 어떠한 영향을 미치며 어떠한 기전으로 해독되는지는 전혀 알려지고 있지 않다. 본 실험연구는 가토와 백서를 대상으로 Flubendazole을 투여하여 실험동물의 간에 어떠한 영향을 미치는지에 대해 조직학적, 조직화학적 및 전자현미경적관찰을 시행함과 동시에 혈청내 효소 및 담취액의 배설능력을 측정하였다. 본 실험은 체중 2kg 내외의 가토 45(정상대조군 18, 실험군 27) 마리와 체중 200g 내외의 백서 37(정상대조군 16, 실험군 21)마리를 대상으로 약물투여 실험을 하였으며, 백서17(대조군 5, 실험군 12)마리를 대상으로 담취액분비를 측정하였다. 가토와 백서의 정상대조군은 24시간 금식후 재급식하면서 실험군외 도살시기에 마추어각각 2마리씩 도살하였다. 약물투여 실험군은 24시간 금식후 Flubendazole의 LD^^50 (2560mg/kg)을 투여하고 재급식하면서 가토는 1, 3, 6, 12, 24시간 및 2, 3, 5, 7일 후에, 백서는 1, 3, 6, 12, 24시간, 2일 및 7일 후에 각자 3마리씩 도살하였다. 정상대조군 및 약물투여군은 도살직전에 채혈하여 가토는 자동분석기로, 백서는 Bessey-Lowry씨법 및 Sigma manual에 의해 alkaline phosphatase, SGOT 및 SGPT치를 측정하였다. 광학현미경적 검사를 위해 정상대조군과 약물투여군의 간을 적출하여 H-E 염색, PAS 염색, D-PAS 염색 및 oil red-0 염색을 시행하였고, 전자현미경적 관찰을 위해서는 조직절편을 3% glutaraldehyde용액으로 고정하고 1% 0^^2 0^^4 용액으로 재고정한 다음 배수알 콜로 탈수하고 Epon 812에 포매한 후 ultramicrotome으로 세편을 만들어 uranyl acetate와 lead citrate로 염색하고 전자현미경으로 관찰하였다. 담취액 분비측정은 정상대조백서군은 마취하여 담취관으로부터 2시간 동안 담취액을 수집하여 그 양을 측정하였고 약을 투여 백서군은 Flubendazole의 LD^^50을 투여한 후 3시간 및 24시간 후에 담취액을 수집 측정하였다. 이상의 실험결과를 요약하면 다음과 같다. 1. Flubendazole LD^^50 (2560mg/kg) 투여군에서는 광학현미경적 소전상 가토 및 백서에서 다 같이 약물투여 3시간 후부터 간세포의 혼탁종창을, 6시간 후부터 공포성변성을관찰하였으며, 이 변화는 약물투여 24시간 후에 최고도에 달해서 48시간 후부터는 감소되 어 5일내지 6일에 정상상태로 회복되는 것을 관찰하였다. 2. 가토에서는 공포성변성이 주로 간소엽 중심부 및 중간부에 나타났고 조직화학적 소견상 그 본체가 주로 당원침착이었던데 비해 백서는 공포성변성이 주로 간소엽주변부 및 중간부에서 관찰되었고 그 본체는 대부분이 지질축적이었다. 3. 전자현미경적 소견상 가토는 정상대조군에 비해 약간의 당원 및 지질축적외에는 별변동이 없었으나 백서에서는 미토콘드리아의 변성, RER의 배열변화, SER의 증가, 다량의지질축적, 세담관의 협착 및 microvilli의 변성이 관찰되었다. 이 변화는 약물투여 3시간후 부터 나타나 12시간후에서 24시간까지가 가장 심했고 48시간후 부터는 회복되는 것을 관찰하였다. 4. Alkaline phophatase, SGOT, SGPT치를 측정했던바 가토에서는 정상대조군과 약물투여군간에 유의한 차가 없었으며 백서에서는 약물투여 6, 12, 24시간 후에 alkaline prosphatase와 SGOT치가 증가하는 경향을 보였다. 5. 백서에서의 담취액분비는 약물투여 3시간후에 대조군에 비해 유의하게 감소했으나 24시간 후에는 유의한 변화가 없었다. 이상의 결과로 보아 본 약제는 특히 백서 실험군에서 투약 후 빠른시기에 일시적으로당원침착, 지질축적을 유발하고 세담관의 변화를 초래하며 담취액배설을 억제하는 것으로 생각된다. [영문] Mebendazole, a derivative of benzimidazole, has been reported as an effective anthelmintic, especially against nematodes, with minimal toxic effects. Nevertheless, due to its poor absorbabil ity through intestinal wall it has been recognized that the compound might not be effective to tissue parasites. Recently another benzimidazole derivative, newly synthesized parafluor analogue of Mebendazole, Flubendazole (Methyl N-[5-(6)-(p-fluorobenzoyl)-2-benzimidazoly] carhamate) was developed from Janssen Pharmaceutica Company. It is white colored, almost tasteless and oderless powder and absorbable through intestinal tract. It has been reported to have broad spectrum anthemintic effect on nematodes and also against certain helminthes in body tissues, such as Clonorchis sinensis in experimentally infected rabbits (Soh et at, 1977). It was suggested that the drug had anthelmintic effect interferring metabolism in cuticle of Clonorchis sinensis resulting a deterioration to protoplasmic tubule (Soh & Min, 1977). However it was found that it had dramatic effect against Clonurchis sinensis in the experimentally infected rabbits but not in rats(Soh et al, 1977). The result suggested that different response might be happened to the drug by animal. At present the pharmacological action or toxic effects of the derivative to animals or human host is not known yet. The present study is purposed to clearify the reason of different effects on clonorchiasis between rabbits and rats and to study the toxic effect of drug to the liver of both animals. In experiments 45 adult rabbits (body weight; around 2kg) and 54 adult rats (body weight; around 200g) were used and devided into 3 groups. In first group (18 normal and 27 drug administered rabbits), the animals were fasted for 24 hours before start of the study and then sacrificed along time schedule (1hr, 3hrs, 6hrs, 12hrs, 24hrs, 2days, 3days, 5days, 6days, and 7days) after administering dose of LD^^50 (2560mg/kg) with feeding. The second rat group (16 normal and 21 drug administered rats) was also fasted for 24 hours and then were sacrificed along time schedule as in the first group. Histological and electron microscopic studies on the liver of the sacrificed animals were done. Assays of serum alkaline phosphatase, SGOT and SGPT were done before sacrificing animals. The third group (17 rats) was provided to test bile flow in rate. Five among them were the normal controls and others were administered with Flubendazole LD^^50 (2560mg/kg). Bile collection for two hours through canula inserted into pancreatico-biliary duct of rat was done under anesthesia in the normal control and 3 hours and 24hours after drug administration. The results are summarized as follows: 1. In light microscopic findings cloudy swelling and vacuolar changes were found from 3 hours and 6 hours after drug administration. The vacuolar changes were on peak 24 hours after drug adminiastration and were recovered as normal 5 days or 7 days after drug administration. These findings were similar in both rabbits and rats. 2. In rabbits the vacuolar changes were observed largely in the centrolobular and midzonal area of the hepatic lobule and their contents were mainly glycogens by histochemical findings. Otherwise in rats they were observed largely in the periportal and midzonal area of the hepatic lobule and the contents were mainly lipids. 3. In electron microcopic findings in rabbits there was no specific alteration except mild deposition of glycogens and lipid droplets in hepatocytes. But in rats degenerative changes in mitochondria, derangement of RER, slightly increased number of SER and lysosomes, deposition of large number of lipid droplets and distorted compact and/or denatured microvilli in bile canaliculi were observed. These changes began 3 hours after drug administration and reached on peak from 12 to 24 hours. They were nearly recovered 6 days after drug administration. 4. No specific difference in the levels of serum alkaline phosphatase, SGOT and SGPT was found in rabbits between the normal control and drug administered groups. But in rats alkaline phosphatase and SGOT showed the tendency of increase at 6, 12 and 24 hours after drug administration. 5. Bile flow in rats was significantly decreased 3 hours after drug administration but not changed 24 hours later. Through above results, it is concluded tbat livers of rat and rabbit show different response to Flubendazole. In rat, more serious damages were appeared in bile canaliculi and more degenerative fatty changes of liver cells were observed.restrictio