Morphine Hydrochloride 및 Demerol Hydrochloride 투여로 인한 백서 장간막 비만세포의 Degranulation 에 관한 조직학적 연구

Abstract

[한글]

[영문]

Introduction

The mast cells were so named by Ehrlich(1879), and many investigators have studied to make a distinction between the activities of the mast cell as a whole and the physiological activities and relations of its granules. Mast cells occur in almost all structures in the body, especially, in loose or areolar connective tissues. Riley(1955) believed that he distinguished two types of mast cells which were distributed in connective tissue, and in blood mast cells called basophils or

mast leukocytes derived from bone marrow.

The metachromatic granules contained in mast cells, were readily diffused from the cells. The physiological implications relative to distribution of mast cells are important because mast cells are considered to be a form of storage of component for, or a readily available supply of, heparin(Holmgren and Wilander, 1937), hyaluronic acid(Asboe-Hansen, 1952), histamine(Riley and West, 1953), serotonin(Benditt et al. 1955), other polysaccharides and certain enzymes or its precursors.

The mode of action of histamine releasers is not clearly understood and, to make the problem more complicated, histamine is present in more than one form and in more than one type of the cell or situation; and it is released by almost any injury or damage to tissues, including the effects of x-rays, ultraviolet or actinic solar rays, electric current, frostbite, and burns.

Maclntosh and Paton(1949), and Paton(1951) suggested that a number of widely different chemical compounds have the common property of releasing histamine from mammalian tissue without producing gross structural change. These substances were called "histamine liberators".

Alam et al.(1939), Rocha and Schild(1949), Mongar and Whelan(1953), Fawcett(1954 a.) Riley and West(1955), and Perry(1956) believed that the ability of compound 48/80 or various chemical histamine liberators to release histamine in experimental animals would appear to depond on a direct interaction and some component of the tissue themselves.

It is well known that the histamine liberated by histamine liberators, such as compound 48/80, stilbamidine, d-tubocurarine and protamine sulfate, increased capillary permeability, falling the blood pressure, and induced edema. Miles(1951), Miles and Miles(1952) and ?? and Miles(1953) demonstrated when the histamine liberator, compound 48/80, injected intravenously into experimental animals with the vital dye pontamine sky blue in their circulation, the skin rapidly becomes blue, and the intradermal injection of substances that increase permeability of the blood vessels in followed by an accumlation of dye at the site of injection. the is presumably due to the passage of dye-stained plasma into the tissue spaces.

It has been also shown that the opium alkaloids and the morphine derivative, apomorphine, have to be added to the class of histamine liberators. Goodman and Gillman(1941), Feldberg and Paton(1950, 1951), Nasmyth and Stewart(1950), Evans et al.(1952), and Parratt and West(1957) demonstrated that in experimental animal, either opium alkaloids or chemical histamine liberators injected intravenously, caused a fall in arterial blood pressured and increased plasma histamine levels simultaneously.

Fawcett(1954a, 1954b, 1955), and Klaus and Winkelmann(1959) demonstreated that the release of mast cell granules by experimental means is accompanied by the release of histamine. It has repeatedly been shown that injection of histamine liberators, such as compound 48/80, reserpine and polymyxin B. caused the

degranulation of the mast cells associated with the release of histamine in the connective tessue. Lee(1965) described that the adminstration of morphine and meperidine induced vasodilatation due to histamine liberation associated with falling blood pressure.

Based upon the above mentioned research works, it is well known that various histamine liberators induced degranulation and disruption of the mast cells and some of the narcotics released histamine. This study has been carried out to demonstrate the degranlation and disruption of mesenteric mast cells in the rat by means of morphine hydrochloride(morphine HCl) and demerol hydrochloride(demerol HCl) administration associated with histamine liberation.

Materials and Methods.

Tje animals used in this experiment were 72 well developed mature albino rats(20 females and 52 males) weighing approximately 200grams.

The narcotics and dosages given for both experimental and control groups are shown in the following table.

Table. Injection sites and dosages in experimental groups

-------------+-----------------------+----------------------+----------------

Group

Intravenous injection

sol. 0.01ml./rat

-------------+-----------------------+----------------------+----------------

The experimental group having an adrenalectomy prior to demerol HCl injection and its control group were added for the indirect effect of the adrenal gland which might induce the degranulation of the mesenteric mast cell by intraperitoneal and intravenous injections.

The adrenalectomy was performed as the procedure used by Oh et al.(1964) and the adrenalectomized rats were available for use in this experiment two weeks after the operation. The direct local injection of narcotics into the mesentery was carried out as the method applied by Pak et al.(1963).

The rats injected intravenously and locally were sacrificed after 2 hours and those given intraperitoneally, after 4 hours. Absolute methanol was directly infused into the peritoneal cavity through a small lncision of the anterior median abdominal wall of the rat, and then fixed for 20 minutes in situ in order to reduce

the direct mechanical injury to the mesenteric mast cell. A few pieces of the mesentery were carefully excised and stained with Pugh's solution containing toluidine blue.

In the groups of the intravenous and intraperitoneal injection, the degrees of degranulation of mesenteric mast cells were divided into 4 grades by means of Ann(1964) as follows; normal mast cells, slight degranulation of mast cells, moderate degranulation of mast cells, and severe degranulation or disruption of mast cells.

In the groups of the local injection, the degranulation of mesenteric mast cell which occurs adjacent to the injected site was carefully studied.

The groups for the potency of morphine HCI and demerol HCI as histamine liberatorys by the intradermal injection in rats, was studied by the method of Lee et al.(1960).

Results

The experimental results are summarized as follows:

1. The experimental groups for intravenous injection.

a. The experimental group for intravenous injection of morphine HCl.

In this group there was a significant decrease of the normal mesenteric mast cell in the incidence of 86.7±3.4%(Mean±Standard error %) of the normal one compared

with the result of the control group occurring about 98.3±0.2% of the normal mast cell. Additionally it may indicated that morphine HCl has a degranulating action on the mast cell of the mesentery with concomitant increase in the atypical or deformed mast cell after intravenous injection of morphine HCl.

b. The experimental group for intravenous injection of demerol HCl.

There was a significant decreased of number of normal mast cells in the incidence of 76.9±11.3% compared with the following result of the control group. Compared to the former group, the degree of degranulation of the mast cell due to demerol HCl

administration seems to be more sever than that of the former group.

c. The control group for intravenous injection of normal saline solution.

In this group, the majority of the mast cells were the normal mast cell showing about 98.3±0.2%. The slight damage or degranulation of mesenteric mast cells were considered due to the stress effect of the injecting procedure without anesthesia.

2. The experimental groups for intraperitoneal injection.

a. The experimental group for intraperitoneal injection of morphine HCl.

In this group there was a slight decrease of the number of normal mesenteric mast cells in the incidence of 90.1±3.9% of normal compared with the incidence of the control group occurring about 95.7±2.0% of the normal mast cell. Compared with the

group for intravenous injection of morphine HCl the degree of the degranulation of the mast cell was slight.

b. The experimental group for intraperitoneal injection of demerol HCl.

After the adiministration of demerol HCl, there was a significant decrease of the normal mast cell in the mesentery with the incidence of 77.9±5.1% normal compared with that of the control group occurring about 95.7±2.0% of the normal mesenteric

mast cell. As the group for intravenous injection of morphine HCl, it indicated that demerol HCl has a degranulating action upon the mast cell associated with the occurrence of atypical or deformed mast cells after intraperitoneal injection of demerol HCl.

c. The control group for intraperitoneal injection of Tyrode solution.

In the control group, the majority of the mesenteric mast cells were the normal mast cell occurring about 95.7±2.0%. The little damage or degranulation of the mesenteric mast cell was considered due to the excess volume of the Tyrode solution

and the same kind of stress mentioned already.

3. The experimental groups for local injection of the mesentery.

a. The experimental group for local injection of morphine HCl.

the mast cells near the injecting site of the mesentery showed morphological changes of slight degranulation and deformity of the cell compared with the normal mesenteric mast cell, while the mesenteric mast cell far from the injection site of morphine HCl showed no morphological changes at all. These findings suggest that morphine HCl effects the mast cell directly or indirectly and cause degranulation of th cell.

b. The experimental group for local injection of demerol HCl.

At the injecting site of demerol HCl is such a small amount no cytological changes of the mast cells were observed. This finding indicated that demerol HCl does not effect mesenteric mast cell directly or indirectly in such dosage of demerol, HCl.

c. The control group for local injection of normal saline solution.

The rats given the normal saline solution locally showed no cytological change at all.

4. The experimental groups having an adrenalectomy prior to demerol HCl injection.

a. The experimental group having an adrenalectomy prior to demerol HCl injection intravenously.

In this group, normal mesenteric mast cells occurred in the incidence of 92.7±1.3% compared with the result of the demerol HCl group give intravenously which was about 76.9±11.3% of normal. This finding indicated that an adrenalectomy prior to demerol HCl injection acted as an inhibitory of protective factor on the degranulation or disruption of mesenteric mast cell of the rat. The result of 92.7±1.3% of the normal mast cell in this group was not significantly different from that of the following control group which was about 93.5±1.5% of normal.

According to the latter finding that shows no significant difference between the result of the adrenalectomized group given demerol HCl interavenously and that of the following control group, no effect of the adrenal gland due to the administration of demerol HCl was noted.

b. The control group having an adrenalectomy prior to normal saline solution injection intravenously.

In this control group, the normal mesenteric mast cell was in the incidence of 93.5±1.5%. The little damage or degranulation of the mast cell in the adrenalectomized rat was considered due to stress at the time of the injecting procedure which was performed without any anesthetic.

c. The experimental group having an adrenalectomy prior to demerol HCl injection intraperitoneally.

In this group the incidence of the normal mast cell was 92.6±2.0% compared with the incidence of the normal one occurring about 9.6±2.6% in this control group. There was no significant difference between the experimental and control groups. This indicated that the adrenalectomy has no effect on the degranulating action of demerol Hcl.

d. The control group having an adrenalectomy prior to Tyrode solution injection intraperitoneally.

In this group the incidence of the normal mast cell was 96.2±2.6%. The slight damage or degranulation of the mast cell was considered due to the same factors mentioned already in the control group for intraperitoenal injection of Tyrode solution.

5. The experimental groups for the permeability of dermal capillaries by morphine HCl demerol HCl.

a. The group for the permeability of dermal capillaries by morphine HCl.

In this experiment the dermal bluing at the injecting sites of histamine and morphine HCl were seen while no dermal reaction at the injection site of normal saline solution was found. According to these findings including the following control experiment give antihistamine prior to the dermal injection of histamine, morphine HCl and normal saline solution, it was proven that morphine HCl liberated tissue histamine as well as histamine injection produced the dermal bluing due to increased permeability of dermal capillaries.

b. The group for the permeability of dermal capillaries by demerol HCl.

No dermal bluing at the injecting sites of demerol HCl and normal saline solution were seen except the site of histamine. These findings indicate that the dermal injection of demerol HCl and normal saline solution did not liberate tissue histamine.

c. The group for the permeability of dermal capillaries by morphine HCl pretreatment with an antihistamine.

In this control experiment the dermal bluing at all injecting sites of mophine HCl, histamine and normal saline solution were did not occur. Additionally the mechanism of an antihistamine was presented by Dale(1950) and Maynard et al.(1955)

while it is not clearly understood.

Summary and Conclusions

Histological studies on the dergranulation of mesenteric mast cells of albino rats caused by injections of morphine HCl and demerol NCl intravenously, intraperitoneally, and local injection of the mesentery of the rats were carried out and the following conclusions were made.

1. In the groups of intravenous, intraperitoneal, and local injections of morphine HCl, the fairly significant degranulation of mesenteric mast cells were observed, which was probably associated with the concomitant liberationof tissue histamine partly derived from the mast cell.

2. In the group of intravenous and intraperiotoneal injections of demerol HCl, relatively significant degranulation of the mesenteric mast cell was recognized. However, the4 local injections showed no cytological change of the mesenteric mast

cell and no liberation of tissue histamine was observed. These findings indicate that the degrnulation of the mesenteric mast cell was caused by a indirect effect due to demerol HCl.

Consequently it is histologically and physiologically deduced that the mode of acting mechanism of morphine HCl and demerol HCl ion the mesenteric mast cell is different.

1ml./rat

normal saline

group

Tyrode sol. 10ml./rat

normal saline sol.

sol. 0.5mg./kg.

control

sol. 12.5mg./kg.

saline sol. 2.4mg./kg.

demerol HCl in

emerol HC in Tyrode

demerol Hcl in normal

sol. 0.5mg./kg.

demerol

normal saline

sol. 50mg./kg.

saline sol. 12mg./kg.

morphine HCl in

group

morphie HCl in Tyrode

morphine HCl in normal

-------------+-----------------------+----------------------+----------------

morphine HCl

injection

Local Injection

Intraperitoneal