급성췌장염의 발생인자에 관한 실험적 연구

Abstract

[한글]

Elucidation of factors in the production of acute pancreatitis

Dong Shik Chin, M.D.

Departments of Pediatrics and Pharmacology, Yonsei University college of

Medicine, Seoul, Korea

(Directed by Drs. T.S. Cho, W.C. Lee, and S.S. Hong)

This paper deals with the effect of intraductal instillation of various agents

including the pancreatic microsome fractions to elucidate a possible factor

concerning the pathogenesis of acute pancreatitis.

The rabbits weighing approximately 2.0kg, regardless of the sex, were used and

divided into following six groups:

1) Intraductal instillation of bile, 1% taurocholate and desoxycholate, and

complete ligation of the pancreatic duct;

2) Intraductal instillation of 20% ethanol or methanol, and complete ligation of

the pancreatic duct;

3) Intraductal instillation of N/10 hydrochloric acid and complete ligation of

the pancreatic duct;

4) Intraductal instillation of 1 μ/cc enterokinase and complete ligation of the

pancreatic duct;

5) Intraductal instillation of pancreatic or myocardial microsome fractions and

complete ligation of the pancreatic duct;

6) Simple ligation of the pancreatic duct of intraductal instillation of saline

and complete ligation of the pancreatic duct, as a control group.

Technic of infusion into pancreatic duct: The abdomen of the rabbit was opened

through an upper midline incision under pentobarbital anesthesia, aseptic technic

was used throughout the operative procedure. The major pancreatic duct, which

enters into the duodenum at a point approximately 20 cm from the pylorus, was

easily cannulated in its intramural potion with a small polyethylene tube. With a

syringe 2.0ml of each prepared solution was infused slowly at a pressure below 30

cm of water. Then, cannula was removed after ligation of the pancreatic duct, and

the abdominal incision was closed.

The serum amylase activity was measured by Nelson's method and serum lipase

activty by Cheery and Crandall's method

Histopathologic examination were carried out on routine hematoxylin-eosin stained

section after 10% formalin fixation and paraffin embedding.

Results and Summary:

1. Instillation of desoxycholate, taurocholate, ethanol or hydrochloric acid into

pancreatic duct following complete ligation in rabbits resulted in marked elevation

of both serum amylase and lipase above the preoperative levels. The maximum rise in

both enzymes usually occurred on the first postoperative day and returned to

preoperative levels on the third day in the case of lipase and on the fourth day in

the case of serum amylase. Thereafter, these enzyme levels were stabilized and did

not vary significantly throughout the duration of the experiment. The slight

elevation of the serum enzymes in rabbits having intraductal methanol, enterokinase

of microsome fractions following complete ligation of the duct is observed. This

change of enzyme value is somewhat similar to those seen in group of simple

ligation or saline instillation. It was also noticed that the levels of serum

enzymes were paralleled with the histologic lesions in the pancreas.

2. Histological examination revealed severe parenchymal necrosis of pancreas in

animals receiving intraductal instillation of bile, desoxycholate. ethanol or

hydrochloric acid. Instillation of taurocholate or enterokinase into the pancreatic

duct following complete ligation induced fat necrosis with marked neutrophilic

infiltration in the pancreas, however, no parenchymal necrosis is observed except a

mild focal necrosis of parenchyma I a part of each group. None of animals showed

parenchymal necrosis in the pancreas following instillation of methanol into the

duct. Moderately severe parenchymal necrosis was noted in animals receiving

pancreatic microsome fraction, whereas, similar treatment with myocardial microsome

fraction failed to produce parenchymal necrosis. After ligation alone or with

intraductal instillation of saline, pancreatic changes were analogous to those seen

in acute edematous or interstitial pancreatitis.

Histologic pictures of animals sacrificed at 4 days or more after procedure

showed more or less similar in all experimental groups, namely acinar atrophy with

ductal transformation and dilation of ductules likely as the effect of ligation,

accompanied by fibroblastic proliferation and chronic interstitial inflammatory

reaction similar to those of chronic pancreatitis in human.

From above results acute necrotizing pancreatitis following instillation of bile,

desoxycholate, ethanol or hydropchloric acid seems to be elicited by their inherent

toxicity to the pancreatic tissue other than activation of proteolytic enzymes,

since it is a well known fact that the pancretic enzymes are not activated, but

rather inhibited by those, particularly by alcohol or hydrochloric acid. On the

other hand, enterokinase treatment following ligation of the pancreatic duct failed

to produce the severe parenchymal necrosis in spite of its activating activity to

proteolytic enzymes. With intraductal instillation of pancreatic microsome fraction

following ligation of the pancreatic duct a moderate degree of parenchylmal

necrosis was resulted. This fact, together with the concept of Reid et al. that

intracellular enzymes might be causing the pancreatic lesion, given rise to the

interesting speculation that the severe pancreatitis with parenchymal necrosis may

be caused by the release of intracellular enzymes, particularly the enzyme or

enzyme system in microsome faction resulting from an admixture of pancreatic juice

and bile or alcohol. Such situation, however, is not induced in case of mild

pancreatitis.

[영문]

This paper deals with the effect of intraductal instillation of various agents including the pancreatic microsome fractions to elucidate a possible factor concerning the pathogenesis of acute pancreatitis.

The rabbits weighing approximately 2.0kg, regardless of the sex, were used and divided into following six groups:

1) Intraductal instillation of bile, 1% taurocholate and desoxycholate, and complete ligation of the pancreatic duct;

2) Intraductal instillation of 20% ethanol or methanol, and complete ligation of the pancreatic duct;

3) Intraductal instillation of N/10 hydrochloric acid and complete ligation of the pancreatic duct;

4) Intraductal instillation of 1 μ/cc enterokinase and complete ligation of the pancreatic duct;

5) Intraductal instillation of pancreatic or myocardial microsome fractions and complete ligation of the pancreatic duct;

6) Simple ligation of the pancreatic duct of intraductal instillation of saline and complete ligation of the pancreatic duct, as a control group.

Technic of infusion into pancreatic duct: The abdomen of the rabbit was opened through an upper midline incision under pentobarbital anesthesia, aseptic technic was used throughout the operative procedure. The major pancreatic duct, which enters into the duodenum at a point approximately 20 cm from the pylorus, was

easily cannulated in its intramural potion with a small polyethylene tube. With a syringe 2.0ml of each prepared solution was infused slowly at a pressure below 30 cm of water. Then, cannula was removed after ligation of the pancreatic duct, and

the abdominal incision was closed.

The serum amylase activity was measured by Nelson's method and serum lipase activty by Cheery and Crandall's method

Histopathologic examination were carried out on routine hematoxylin-eosin stained section after 10% formalin fixation and paraffin embedding.

Results and Summary:

1. Instillation of desoxycholate, taurocholate, ethanol or hydrochloric acid into pancreatic duct following complete ligation in rabbits resulted in marked elevation of both serum amylase and lipase above the preoperative levels. The maximum rise in both enzymes usually occurred on the first postoperative day and returned to preoperative levels on the third day in the case of lipase and on the fourth day in the case of serum amylase. Thereafter, these enzyme levels were stabilized and did

not vary significantly throughout the duration of the experiment. The slight elevation of the serum enzymes in rabbits having intraductal methanol, enterokinase of microsome fractions following complete ligation of the duct is observed. This

change of enzyme value is somewhat similar to those seen in group of simple ligation or saline instillation. It was also noticed that the levels of serum enzymes were paralleled with the histologic lesions in the pancreas.

2. Histological examination revealed severe parenchymal necrosis of pancreas in animals receiving intraductal instillation of bile, desoxycholate. ethanol or hydrochloric acid. Instillation of taurocholate or enterokinase into the pancreatic duct following complete ligation induced fat necrosis with marked neutrophilic infiltration in the pancreas, however, no parenchymal necrosis is observed except a mild focal necrosis of parenchyma I a part of each group. None of animals showed

parenchymal necrosis in the pancreas following instillation of methanol into the duct. Moderately severe parenchymal necrosis was noted in animals receiving pancreatic microsome fraction, whereas, similar treatment with myocardial microsome

fraction failed to produce parenchymal necrosis. After ligation alone or with intraductal instillation of saline, pancreatic changes were analogous to those seen in acute edematous or interstitial pancreatitis.

Histologic pictures of animals sacrificed at 4 days or more after procedure showed more or less similar in all experimental groups, namely acinar atrophy with ductal transformation and dilation of ductules likely as the effect of ligation,

accompanied by fibroblastic proliferation and chronic interstitial inflammatory reaction similar to those of chronic pancreatitis in human.

From above results acute necrotizing pancreatitis following instillation of bile, desoxycholate, ethanol or hydropchloric acid seems to be elicited by their inherent toxicity to the pancreatic tissue other than activation of proteolytic enzymes,

since it is a well known fact that the pancretic enzymes are not activated, but rather inhibited by those, particularly by alcohol or hydrochloric acid. On the other hand, enterokinase treatment following ligation of the pancreatic duct failed to produce the severe parenchymal necrosis in spite of its activating activity to proteolytic enzymes. With intraductal instillation of pancreatic microsome fraction following ligation of the pancreatic duct a moderate degree of parenchylmal

necrosis was resulted. This fact, together with the concept of Reid et al. that intracellular enzymes might be causing the pancreatic lesion, given rise to the interesting speculation that the severe pancreatitis with parenchymal necrosis may

be caused by the release of intracellular enzymes, particularly the enzyme or enzyme system in microsome faction resulting from an admixture of pancreatic juice and bile or alcohol. Such situation, however, is not induced in case of mild pancreatitis.