抗生劑(Chloramphenicol 및 Neomycin)가 肝 및 其他臟器에 미치는 影響에 關한 組織化學的 硏究
(A) Histochemical study on the effects of chloramphenicol and neomycin in the livers and the other organs of
Since the structural formula of chloramphenicol was determined, its chemistry, pharmacology, and clinical usage were extensively studied by many researchers.
Recently more and more cases of gram-negative sepsis are being recognized and treated, and there is a tendency to use chloramphenicol in large doses when a case of gram-negative infection is suspected. Since 1950, awareness is increasing that
patients with cirrhosis of the liver are susceptibel to gram-negative septicenia, and diabetics are known to be especially susceptible to the disorder. Lazar(1966) emphasized that due care should be taken in the administration of chloramphenicol in such cases. He has reported the development of toxicity of cholramphenicol in the case of hepatic cirrhosis with gram-negative sepsis. Kunin et al.(1959) have shown that intoxication developed easily in patients with hepatic or renal diseases
treated with chloramphenicol.
The effect of chloramphenicol on protein and RNA syntheses in bacteria and mammalian cell suspensions has been studied by many investigators, and some reported that the syntheses were inhibited, although others observed no inhibition of the syntheses by low concentrations.
Gale and Folkes(1953) reported that in tissue cultures chloramphenicol inhibited protein synthesis. Lepage(1953) reported that in ascites tumor-cell-suspensions chloramphenicl inhibited a certain incorporation of RNA precursor. On the contrary, Kapool et al.(1963), von Ehrenstein and Lipmann(1961), Ren야 and Ochoa(1962), and Allen and Schweet(1962) have reported that protein synbesis was not inhibited by chloramphenicol in the cell-free system of the vertebrate cells.
Since Peck et al.(1949) purified neomycin and found it to be a complex of three compounds, neomycin A,B, and C with different antimicrobial activities, it is known that neomycin has a broad spectrum and is effective against gram-negative, gram-positive organisms, and against tubercle bacilli. It has clinical usefulness as an intestinal antiseptic prior to bowel surgery.
Emmerson and Davies(1964) indicated that administration of neomycin may produce considerable change in the proximal convoluted uriniferous tubules, especially enzymatic decrease of alkaline phosphatase and lactate dehydrogenase. Gawecka et
al.(1966) found that long-term administration of neomycin exerted a nephrotoxic influence. Sazykin and Dhernukh(1963) observed that antibiotics of the neomycin group added to bacteria cultures had interrupted protein synthesis.
The author has attempted this histochemical study in order to observe the effects of large doses of chloramphenicol and neomycin on the livers, kidneys and spleens of mice by observing the changes in several enzymes and nucleic acid correlated
with protein synthesis.
MATERIALS AND METHODS
Seventy-two adult finale mice were divided into five groups as follows.
A) Experimental groups.
Group Ⅰ. acute toxicity with chloramphenicol (Chloramphenicol sodium succinate, Parke Davis & Co.), 1.32mg/g/day.
Group Ⅱ. chronic toxicity with chloramphenicol, 0.25mg/g/day.
Group Ⅲ. acute toxicity with neomycin(mycifradin sulfate, Upjohn), 0.13mg/g/day.
Group Ⅳ. chronic toxicity with neomycin, 0.25mg/g/day.
B) Control group.
Group Ⅴ. physiological saline solution.
For acute toxicity to the experimental groups, the drugs were intraperitoneally injected twice in a day, and for chronic toxicity groups, also intraperitoneally twice daily for 14days.
All small pieces of the livers, kidneys and spleens were freshly obtained, and fixed or unfixed, and then the following procedures were carried out.
A modified technique of Gomori's method was applied for the demonstration of acid phosphatase and alkaline phosphatase activities. The method presented by wachstein and Meisel(1957) was used for the activities of adenosine diphosphatase(ADEase) and adenosien triphosphatase (ATPase). The activity of succinic dehydrogenase was demonstrated by the method of Nachlas et al.(1957). The Periodic acid-Schiff method (Hotchkise, 1948) for polysaccharides and the Methyl green-Pyronin stainig (Rosa,
1950) for nucleic acids, and Hematoxylin-Eosin staining were respectively used.
SUMMARY AND CONCLUSION
The activities of acid phosphatase, alkaline phosatase, adenosine diphosphatase, adenosine triphosphatase, succinic dehydrogenase, and the changes of nucelic acid and polysaccharides in the livers, kidneys, and splees were histochemically observed in healthy male mice treated which large doses of chloramphenicol and neomycin.
1. In the acid phosphatase activity, some or no changes were found in the Kupffer cells of the liver. The enzymatic activities of the hepatic cells were generally reduced, especially noted in the neomycin groups.
2. In the alkaline phosphatase, enzymatic activities of the specimens of the experimental groups were reduced, Marked decrease of the enzymatic activities of the splenic tissue incases of acute chloramphenicol intoxication group was observed
compared with the control group.
3. The ADpase and ATPase activities were reduced in the hepatic cells, the bile canaliculi, and the vascualr and sinusoidal walls. Slight or no changes were found in the renal and splenic tissues.
4. Succinic dehydrogenase activities of the hepatic tissues were generally reduced, especially marked in the intermediated zone of the hepatic lobule. General decrease in enzymatic activities was observed in the renal and splenic tissues.
Inactive areas were easily found in the renal and splenic parenchymal tissues.
5. In the changes of the nucleic acid and the polysaccharides, pyroninophill substances were generally reduced in the hepatic tissues, and PAS-positive substances were also reduced in all of the experimental groups.
It is suggested that the antibiotics chloramphenicol and neomycin have a suppressing effect on enzymatic activities in livers, kidneys, and spleens of mice and also on protein synthesis.