Cortisone 투여가 간디스토마 감염숙주와 蟲體에 미치는 영향

Abstract

[한글]

[영문]

Kass & Finland(1953) have reported the evidence for depression of resistance to infection following the administration of ACTH and cortisone which is due to the adverse effect of these on antibody production and phagocytic activity.

The results of the experiments of Weinstein(1953) with Nippostrongylus muris, of Roman(1956) with Strongylodies ratti, of Cross(1960) with Nematospiroides dubius, and Mathis(1962) with Aspicularis tetraptera in cortisone-treated rats and mice

indicated that cortisone depressed the resistance to Nematode infection. Likewise, in Trematode infection, Lee(1967) and Ohda(1967) reported the same increased susceptibility to the Clonorchis and Paragonimus infection following administration

of cortisone.

As to the mechanism for the effect of cortisone on parasitic infection, Lindquist(1950) and Coker(1956) found that cellular response in important in the best defence and this response was suppressed by cortisone.

Lewart & Lee(1954) reported the reduction of immunity against larva following the administration of cortisone.

They gave cortisone to a host infected with schistosomes. Resistance, which was derived from the local tissue protein, against the larval invasion was reduced.

Tsuchi(1961) reported the reduction of tissue stain with Evans blue after continuous cortisone administration.

Inflammatory reaction was depressed due to inhibition of the metabolism of glycoprotein and polysaccharides.

Lindquist(1950) indicated that a fibrotic encapsulation of the worm found at the time of the larval migration was one of the cellular responses which inhibited parasite growth. Lee(1967) reported failure of encapsulation in Paragonimus infections after cortisone.

The present investigation studies the effects of cortisone on the host and parastie of clonorchis sinensis at the different developmental stages, using various scheduled for cortisone administration.

Materials and Methods

Animals : Sixty-four healthy white rabbits(1.8-2.6kg) were used for the experiment.

Metacercariae: Metacercariae of Clonorchis sinensis from fresh water fish caught at the endemic area, were collected by the digestion method using artificial gastric juice.

Grouping of the rabbits and the administration of cortisone were as follows;

Grpoup Administation schedule

Ⅰ.Control group

Ⅰ-A Normal

Ⅰ-B Cortisone only, long term

Ⅰ-C Infection, without cortisone

Ⅱ. Experimental group

Ⅱ-A^^1 Cortisone from 1 week before the infection, long term

Ⅱ-A^^2 Cortisone from the time of the infection, long term

Ⅱ-B^^1 Cortisone only for 1 week before the infection, short term

Ⅱ-B^^2 Cortisone only for 1 week from the time of the infection, short term

Ⅱ-C^^1 Cortisone from 30th day after the infection, long term

Ⅱ-C^^2 Cortisone from 64th day after the infection, long term

Cortisone acetate(Keunwha C0.) 15mg/Kg was given by intramuscular injection into

the thigh of the rabbit every day(Ⅱ-A^^1, Ⅱ-A^^2, Ⅱ-B^^1 and Ⅱ-B^^2) or on

every other day(Ⅱ-C^^1 and Ⅱ-C^^2).

Body weight: Every three days animals were weighed in a fasting state in the morning.

Liver function test: Serum glutamic oxaloacetic transaminase was measured by following method; Serum(0.1ml) was incubated with 0.5ml of glutamicoxaloacetic transaminase substate for one minute at 37。C, and read by 500mμ juniorspectrophotometer 20 minutes after addition 0.5ml of 2.4 dinitrophenyl hydrazine and 5ml of 0.4 N-NaOH solution.

Serum fraction and A/G ratio: Paper elotrophoresis was used. Serum was run on the filter paper(Whatman No.1) for 16 hours and dyed with bromphenol blue. Each fraction was elutriated by 0.1% Na^^2CO^^3 and read by 520mμ junior spectrophotometer.

Pathological examination: Animals were killed at the end of the experimental period. At sutopsy, liver and adrenal glands were grossly examined, were fixed in 10% of formalin, and cut in 6 micron thickness after paraffin embedding. The tissues were stained with routine hematoxylin-eosin. periodic acid Schiff stain was combined with diastase digestion to measure glycogen.

Results

Effect of cortisone on the growth of adult worams of Clonorchis sinensis: Average body length of worms recovered from the experimental and control rabbits, which were killed 8 weeks after infection, was as follows; average size of the worms detected from cortisone-treated rabbits (Ⅱ-A^^1 and Ⅱ-A^^2 group) were

13.2±0.21mm and 12.6±0.24mm. On the other hand, from Ⅱ-B^^! and Ⅱ-B^^2 group showed 11.0±0.24mm and 11.2±0.26mm, and in control group(I0C0 showed 10.9±0.23mm.

Average size of worms recovered from untreated control rabbis treated by cortisone for a long term was significantly larger than that of worms from untreated control rabbis 8 weeks after the infection.

No significant difference was noted between the short term group and untreated control group.

Effect of cortisone on the infectivity of Clonorchis sinensis: Average number of worms detected in the liver from Ⅱ-A^^1 and Ⅱ-A^^2 group was 38.6±3.95 and 36.8±2.35, 27.8±2.58 in Ⅱ-B^^1, 24.5±3.28 in Ⅱ-B^^2 and 26.2±2.70 worms in untreated control group.

A significant difference of Clonorchis infectivity was noted between the control group and the group treated with cortisone for a long term.

Effect of Cortisone on the body weight of rabbits infected with Clonorchis sinensis: In group Ⅱ-A^^1 and Ⅱ-A^^2 all animals showed a rapid weight decrease, without a progressive weight increase in the later stage. During the period of cortisone administration, especially. loss of weight was more severe in Ⅱ-A^^1 group than those of Ⅱ-A^^2. In group Ⅱ-A^^1 weight was approximately half of initial weight, and dropped to 28% of the initial weight in group Ⅱ-A^^1 on the 8th week of the infection.

In group Ⅱ-B^^1 weight gradually decreased for 2 weeks following one week of cortisone administration. The recovery of weight was slow, reaching initial weight not he 8th week of the infection. In group Ⅱ-B^^2 the weight change was similar to

that of the untreated control group.

Through this observation, in Ⅱ-A^^1 and Ⅱ-B^^1 in those in which treatment was initiated before infection, weight loss was rapid and severe as compared with the change of weight in group Ⅱ-A^^2 and Ⅱ-B^^2.

Weight changes of liver and adrenal glands of cortisone-treated rabbits: There was no significant difference in liver weight or the rabbits between cortisone-treated group and normal control group on the 13th week of the infection. Liver weight/body weight ration showed 3.3-3.0% in group Ⅱ-C^^1 and Ⅱ0C^^2, 3.1% in I-B and normal control group.

Concerning the weight of adrenal gland, there were no significant differences among groups Ⅱ-C^^1, Ⅱ-C^62 and I-A, but in group I-B significant weight reduction was observed.

Effect of cortisone of liver function of rabbits: In group I-A and I-B S.G.O.T. values were normal. In Ⅱ-C^^1 group it was a rapid increase to 114 units on the 40th day of the infection (10days after the initiation of cortisone administration). Then a rapid decrease was observed.

In case of Ⅱ-C^^2 relatively high value appeared at the time of showing peak value in the group Ⅱ-C^^1. These results indicate the high value of S.G.O.T. probably was not due to cortisone administration but rather to the Clonorchis infection.

Effect of cortisone on serum protein fraction and A/G ratio: In I-C group, untreated infection group, significant increase of γ-globulin level and reduetion of A/G ratio were observed. On the other hand, in cortisone-treated group, Ⅱ-A^^1, Ⅱ-A^^2 and I-B, significant differences as compared with initial values were not

observed.

Pathological findings: Histopathologic examination of liver in group I-C showed a moderate adenomatous hyperplasis of the bile ducts with occasional epithelial necrosis, and marked inflammatory changes in the periductal area with fibrosis.

These inflammatory changes extended into the hepatic parenchyma and were similar in degree, but in group Ⅱ-A^^1 and Ⅱ-B^^1, the changes were quite in the portal spaces and hepatic parenchyma. In group Ⅱ-A^^2 and Ⅱ-B^^2 the changes were quite mild and limited to the portal spaces. The hepatic cells, particularoy in groups having prolonged cortisone administration, were swollen and foamy vesicular. PAS stain was positive.

The adrenal glands showed no specific changes except for atrophic changes, particularly in the zona fasciculata, in the groups having prolonged cortisone administration.

Conclusion

The effects of cortisone on the host and parasite of Clonorchis sinensis were studied. Fifteen mg/Kg of cortisone was administrated to Clonorchis-infected ravvits having different developmental stages of the parasite, using various schedules of administration, long or short term.

1. Rapid and severe weight loss was seen in rabbits given cortisone over a long term. There was a longer parasite body length and high hepatic infestation rate in long term group.

2. There was no significant difference in the liver and adrenal gland weights of the rabbits between cortisone-treated group and normal control group. In the group treated by cortisone for long term there was marked reduction in the weight of the adrenal gland. The peak S.G.O.T. value was observed 40days after the infection and had no significant effect on this.

3. As significant increase in γ-globulin and reduction of the A/G ratio was found in untreated control group. In treated group such changes were not observed.

4. The inhibition of inflammatory changes of the liver was noted in rabbits treated by cortisone from the larval stage of infection, but a significant variation was not observed in the group treated at adult stage of the parasite and control group.

From the above it is concluded that cortisone amy cause increased susceptibitlity to infection by Clonorchis sinensis, and may inhibit the hepatic inflammatory reaction. These effects are more significant when the administration of cortisone

is continued on a long term basis beginning at the larval stage of the parasite.