Morphine Hydrochloride 및 Meperidine Hydrochloride가 흰쥐 장간막 비만세포에 미치는 영향에 관한 실험적 연구

Abstract

[한글]

The experimental studies an the effect of morphine and meperidine hydrochorides on

the mesenteric mast cells of albino rats

Hyun Sam Shin

Department of Medical Science

The Graduate School Yonsei University, Seoul, Korea

(Directors: Prof. Kum Duck Choi and Soo Yun Pak)

It is conmon knowledge that tissue most cells contain heparin, histamine,

Hyaluronic acid, serotonin and other components. In cartes of extreme need the

cells will secrete these cyto-plasmic contents. Under certain pathological

condition and by the use of experimental means as, for example, with an injection

of histamine liberator mast cells will release cytoplasmic metachromatic granules

or disrupt readily.

Junqeira and Beiguelman(1955), in vitro, studied the action of compound 48/80, at

low concentrations, promoting a vigorous extrusion of rat's mesenteric most cell

granules. They observed that low temperatures below 25℃, acid pH, and several

metabolic inhibitors prevented granule extrusion which was observed after the

addition of compound 48/80. This consequently suggested that metabolic phenomena

occuring in mast cells are of vast importance considering their response to the

action of compound 48/80.

Lee(1968) and Park et al. (1970) studied the action of morphine and meperidine

hydrochloride by means of intravenous injection. This consequently induced a

degranulatien of rat's mesenteric mast cells. Thusly, it was concluded that the

degranulation of the cells was caused by both a direct and indirect effect. This

was due to morphine hydorchloride and only indirectly was it due to the meperidine

hydrochloride.

The present study of albino rats, in vitro, was undertaken to determine the

effect of morphine and meperidine hydrochlorides and to observe whether or not the

metabolic inhibitor effects the action of narcotics on the degranulation and

disruption of mesenteric mast cells.

Experimental animals used in this study were 70 healthy mature male rats of

Sprague-Dawley strain weighing approximately 200 Gm. each. For the purpose of

studying the effects of morphine and meperidine hydrochlorides on mesenteric mast

cells in vitro, the experimental groups divided as follows.

A. The groups incubated in morphine hydrochloride solution :

a. The experimental group consisting of those incubated at 0.01mg./ml. of

morphine hydrochloride in Tyrode solution.

b. The experimental group consisting of these incubated at 0.02mg./ml. of

morphine hydrochloride in Tyrode solution.

c. The experimental group consisting of those incubated at 0.04mg./ml. of

morphine hydrochloride in Tyrode solution.

d. The control group of Tyrode solution.

B. The groups incubated in meperidine hydrochloride solution :

a. The experimental group consisting of the incubated at 0.02mg./ml. of

meperidine hydrochloride in Tyrode solution.

b. The experimental group consisting of those incubated at 0.04mg./ml. of

meperidine hydrochloride in Tyroide solution.

c. The Control gruop of Tyrode solution.

C. The groups incubated with morphine hydrochloride solution plus iodoacetic acid

:

a. The experimental group consisting of those incubated at 0.02mg./ml, of

morphine hydrochloride plus 0.03×10**-2 M of iodoacetic acid in Tyrode solution.

b. The experimental group consisting of those incubated at 0.04mg./ml. of

morphine hydrochloride plus 0.33×10**-2 M of iodoacetic acid in Tyrode solution.

c. The control group consisting of those incubated at 0.02mg./ml. of morphine

hydrochloride in Tyrode solution.

d. The Control group consisting of those incubated at 0.04mg./ml. of morphine

hydrochloride in Tyrode solution.

D. The adrenalectomized groups incubated in morphine hydrochloride solution :

a. The adrenalectomized group consisting of those incubated at 0.02mg./ml. of

morphine hydrochloride in Tyrode solution.

b. The non adrenalectomized group consisting of those incubated at 0.02mg./ml. of

morphine hydrochloride in Tyrode solution.

c. The adrenalectomized group incubated in Tyrode solution also act as the

control group.

In each experimental group mesenteric parts were carefully excised from the

experimental animals which previously had been sacrified by an occipital blow.

Pieces of the various mesenteries were then immediately incubated in prewarmed

Tyrode solution at 37℃ containing the above mentioned doses of morphine

hydrochloride. meperidine hydrochloride and morphine hydrochloride plus iodoacetic

acid for periods of 10, 20 and 30 minutes respectively. After the incubation the

pieces were fixed in absolute methanol for 20 minutes and then washed lightly in

distilled water.

Each piece was stained in Pugh's solution which formerly was used by LeBlanc and

Rosenberg(1957). A permanent histological slide was prepared for observation using

a light microscope at high power magnification.

For the different degrees of degranulation of mesenteric mast cells 4 grades of

cytological changes were adapted by the criteria of An(1974) as follow :

1. The normal type of mast cell displays mostly a round form without any evident

dispersion of metachromatic granules from the cell. However, the mart cells showing

one or two extracelluar metachromatic granules were also counted as normal types.

(Fig. 1)

2. The grade Ⅰ type showing a slight degranulation of a mart cell with a few to

several metachromatic granules in it'S Vicinity, (Fig. 2)

3. The grade Ⅱ type showing a moderate degranultion of a roast cell whith a

clear contour(Fig. 3)

4. The grade Ⅲ type having a severe degranulation or disruption of the mast cell

in whieh the clear contour of the cell is difficult to identify due to the severe

degranulation or disruption. (Pig. 4)

The adrenalectomy and postoperative care of the rat were performed according to

the procedures previeously used by Oh et al. (1954).

In the experimental groups incubated at 0.01mg./ml. and 0.02mg./ml. of morphine

hrdro chloride in Tyrode solution the degranulation of mesenteric mast cells showed

no significant difference from that of the control group. However. in the

experimental group incubated at 0.04mg./ml. of morphine hydrochloride in Tyrode

solution the degranulation of the cells were somewhat increased over that of the

control group. In vitro, the dose of morphine hydrochloride in Tyrode solution that

induces a degranulation of mesenteric mart cells was found to he 0.04mg./ml. of

Tyrode solution.

In the experimental groups of meperidine hydrochloride in Tyrode solution the

degranulation of mesenteric mast cells showed result similar to that of the control

group. With the dosage of meperidine hydroshloride the cells didn't show an

increased degranulation of metachromatic granules.

By the effect of metabolic inhibiter such as an addition of iodoacetic acid in

the morphine hydrochloride solution the degranulation was evidently inhibited for

20 minutes during the incubation process. Howeyer, for a period of 30 minutes the

dearanulation of mesenteric mast cells was markedly increased as compared with the

result of the control group. This fact was considered as the probable degenerative

change of the cells due to the metabolic inhibitor or iodoacetic acid for 30

minutes period during the process of incuhation.

Consequently the author has demonstrated the effect of morphine hydrochloride in

its ability to induce a degranulation of mesenteric mast cells in vitro.

[영문]

It is conmon knowledge that tissue most cells contain heparin, histamine, Hyaluronic acid, serotonin and other components. In cartes of extreme need the cells will secrete these cyto-plasmic contents. Under certain pathological condition and by the use of experimental means as, for example, with an injection of histamine liberator mast cells will release cytoplasmic metachromatic granules or disrupt readily.

Junqeira and Beiguelman(1955), in vitro, studied the action of compound 48/80, at low concentrations, promoting a vigorous extrusion of rat's mesenteric most cell granules. They observed that low temperatures below 25℃, acid pH, and several metabolic inhibitors prevented granule extrusion which was observed after the addition of compound 48/80. This consequently suggested that metabolic phenomena occuring in mast cells are of vast importance considering their response to the action of compound 48/80.

Lee(1968) and Park et al. (1970) studied the action of morphine and meperidine hydrochloride by means of intravenous injection. This consequently induced a degranulatien of rat's mesenteric mast cells. Thusly, it was concluded that the degranulation of the cells was caused by both a direct and indirect effect. This

was due to morphine hydorchloride and only indirectly was it due to the meperidine hydrochloride.

The present study of albino rats, in vitro, was undertaken to determine the effect of morphine and meperidine hydrochlorides and to observe whether or not the metabolic inhibitor effects the action of narcotics on the degranulation and disruption of mesenteric mast cells.

Experimental animals used in this study were 70 healthy mature male rats of Sprague-Dawley strain weighing approximately 200 Gm. each. For the purpose of studying the effects of morphine and meperidine hydrochlorides on mesenteric mast cells in vitro, the experimental groups divided as follows.

A. The groups incubated in morphine hydrochloride solution :

a. The experimental group consisting of those incubated at 0.01mg./ml. of morphine hydrochloride in Tyrode solution.

b. The experimental group consisting of these incubated at 0.02mg./ml. of morphine hydrochloride in Tyrode solution.

c. The experimental group consisting of those incubated at 0.04mg./ml. of morphine hydrochloride in Tyrode solution.

d. The control group of Tyrode solution.

B. The groups incubated in meperidine hydrochloride solution :

a. The experimental group consisting of the incubated at 0.02mg./ml. of meperidine hydrochloride in Tyrode solution.

b. The experimental group consisting of those incubated at 0.04mg./ml. of meperidine hydrochloride in Tyroide solution.

c. The Control gruop of Tyrode solution.

C. The groups incubated with morphine hydrochloride solution plus iodoacetic acid :

a. The experimental group consisting of those incubated at 0.02mg./ml, of morphine hydrochloride plus 0.03×10**-2 M of iodoacetic acid in Tyrode solution.

b. The experimental group consisting of those incubated at 0.04mg./ml. of morphine hydrochloride plus 0.33×10**-2 M of iodoacetic acid in Tyrode solution.

c. The control group consisting of those incubated at 0.02mg./ml. of morphine hydrochloride in Tyrode solution.

d. The Control group consisting of those incubated at 0.04mg./ml. of morphine hydrochloride in Tyrode solution.

D. The adrenalectomized groups incubated in morphine hydrochloride solution :

a. The adrenalectomized group consisting of those incubated at 0.02mg./ml. of morphine hydrochloride in Tyrode solution.

b. The non adrenalectomized group consisting of those incubated at 0.02mg./ml. of morphine hydrochloride in Tyrode solution.

c. The adrenalectomized group incubated in Tyrode solution also act as the control group.

In each experimental group mesenteric parts were carefully excised from the experimental animals which previously had been sacrified by an occipital blow.

Pieces of the various mesenteries were then immediately incubated in prewarmed Tyrode solution at 37℃ containing the above mentioned doses of morphine hydrochloride. meperidine hydrochloride and morphine hydrochloride plus iodoacetic

acid for periods of 10, 20 and 30 minutes respectively. After the incubation the pieces were fixed in absolute methanol for 20 minutes and then washed lightly in distilled water.

Each piece was stained in Pugh's solution which formerly was used by LeBlanc and Rosenberg(1957). A permanent histological slide was prepared for observation using a light microscope at high power magnification.

For the different degrees of degranulation of mesenteric mast cells 4 grades of cytological changes were adapted by the criteria of An(1974) as follow :

1. The normal type of mast cell displays mostly a round form without any evident dispersion of metachromatic granules from the cell. However, the mart cells showing one or two extracelluar metachromatic granules were also counted as normal types.

(Fig. 1)

2. The grade Ⅰ type showing a slight degranulation of a mart cell with a few to several metachromatic granules in it'S Vicinity, (Fig. 2)

3. The grade Ⅱ type showing a moderate degranultion of a roast cell whith a clear contour(Fig. 3)

4. The grade Ⅲ type having a severe degranulation or disruption of the mast cell in whieh the clear contour of the cell is difficult to identify due to the severe degranulation or disruption. (Pig. 4)

The adrenalectomy and postoperative care of the rat were performed according to the procedures previeously used by Oh et al. (1954).

In the experimental groups incubated at 0.01mg./ml. and 0.02mg./ml. of morphine hrdro chloride in Tyrode solution the degranulation of mesenteric mast cells showed no significant difference from that of the control group. However. in the

experimental group incubated at 0.04mg./ml. of morphine hydrochloride in Tyrode solution the degranulation of the cells were somewhat increased over that of the control group. In vitro, the dose of morphine hydrochloride in Tyrode solution that

induces a degranulation of mesenteric mart cells was found to he 0.04mg./ml. of Tyrode solution.

In the experimental groups of meperidine hydrochloride in Tyrode solution the degranulation of mesenteric mast cells showed result similar to that of the control group. With the dosage of meperidine hydroshloride the cells didn't show an increased degranulation of metachromatic granules.

By the effect of metabolic inhibiter such as an addition of iodoacetic acid in the morphine hydrochloride solution the degranulation was evidently inhibited for 20 minutes during the incubation process. Howeyer, for a period of 30 minutes the

dearanulation of mesenteric mast cells was markedly increased as compared with the result of the control group. This fact was considered as the probable degenerative change of the cells due to the metabolic inhibitor or iodoacetic acid for 30 minutes period during the process of incuhation.

Consequently the author has demonstrated the effect of morphine hydrochloride in its ability to induce a degranulation of mesenteric mast cells in vitro.