Pergamon Press
Printed in the U.S .A .
NALORONE BLOCKADE OF
ACUPUNCTURE ANALGESIA :
ENDORPHIN IMPLICATED
Department of Zoology
University of Toronto
Summary
Methods
1757
1758 Naloxone Blocks Acupuncture Analgesia Vol . 19, No . 11
through holes in the aide of the funnel and were immobilized with masking tape
for later application of acupuncture needles (Control mice not receiving acu-
puncture were immobilized in the same manner) . Squeaks were elicited using
noxious heat from a elide projector lamp (GE-ELH) pointed at the protruding
nose at a distance of 10 cm . The restrained mouse and lamp were in a sound
proof chamber as ambient noises disturbed the animal . The squeak was monitored
through a microphone, headphones, and a storage oscilloscope (Tektronix 5103N)
externally triggered by the lamp switch . The latency of the squeak was measur
ed accurately on the oscilloscope and averaged 3 .0 seconds (S .E . _+ 0 .05) . The
oscilloscope traces were used to exclude small amplitude squeaks which occas-
ionally occurred within 2 seconds of turning on the lamp . These were discarded
from the data as they resembled startle reaponaea with small intensities quite
different from the stressful sounds occurring at 3 .0 seconds . The criteria for
rejecting small early squeaks were any sounds with amplitudes less than one
half that of the true stressful response and occurring within 2 seconds of the
onset of the heat stimulus . Before starting the various treatments, each
mouse was given four control tests, two minutes apart, and the mean latency was
used for zero time control values . We selected those mice which gave 4 reprod-
ucible control reaponaea with latencies between 2 to 4 seconds ; we rejected 5
out of 75 mice . Next, we tested each mouse 6 times over 140 mina .
Reeulta
Figures 1 and 2 and table 1 summarize the averaged data for each group,
giving percentage change in latency to squeak . Figure 1 shows the values
obtained from groups I to IV and figure 2 the data from groups V to VII . The P
values indicated by letters were obtained by comparing the means at a given
time with zero time control values . (All P values in this paper were calcula-
ted from two-tailed student t tests .)
-20
-ao ~+ ~ ±
O 6 20 "0 BO BO 100 120
TIME (MIN .1
TIME (MIN .)
O~ u'1 1~ wM M
~O
ô
N M M M N ~Y
H +~
H
y M M ~t 00 ~t u1
m I I I I I I M
'-I H
M I~ W ~O CO 'i
'-I ~-i r~ r-I O~ u1
O .--I N N ""m O O
3+
~+
H
30 O
C
O O O O W r-1 ~7 +~ P+
V I I I
O ao /
ri W
N W
,au uu1 ,o~ av1 aY aO a
N ^ "o
H N rl N rl N N D
riC ~~ ...
m O~
O "^
u1
L O 00 00 O~ N O~ O N
I I r-I .--I I H aô 0
v I I I
+I
Ha 0o ao a
a aN a~O m co
O~ 00 I~ 'i ~ c~ /
O
u N V1 ~O ~7 M 0 ^ 0
N ~
H +I +I +I +I +I +I H
W N
.-I ON"^ O
W
.-I N ~7 N N ~H O
W O
a V
+I Wô w
C Al Al ÔO Ô~ N 00 v7 +~ II
CI M M M rl M N +~H 21
v H (', M
H
H n 00 v1
u1 ~O
~
O~
H^ Ô
~ H I I 1
~H O
O
m
H
a . b .^ W
it W AO~ Ol w a r+ n
O ic O W N M 00 Ou rl. u
+~
0
m
N N M rl N N 01 O
~, 7 H
H
CN M 1~ I~ q +~ ô
q
d 1~ ~O rl rl O
u D
I I I I I I
mM O
Qi a
a
0 ,-Id ^ ~ v
w
W L~ ,nO N m U p~Y U H .-I
O O N O ~t qH " /
H ~O ~O u'1 00 O 0 O
0
00 W N A
H O~ +I
H 1~ H r1 O O
N
~O M
M d
vl ~t '-I
M ~O
rl roOM O
U L+m
+~H O
000 ,al
.p ao ~I â
û û m
q m v N /
0 N ~t ~O O d a
N â. H
U F
â
Vol . 19, No . 11 Naloxone Blocks Acupuncture Analgesia 1761
for acupunctured mice (P« ,001) and a 44% increase for acupuncture plus saline
(P« ,001) . In addition to these within group statistics, we also did t tests
to compare the percentage changes between pairs of groups at 40 minutes. These
results show the following : First, that sham acupuncture in group II produced
no analgesia, showing that group I had a true acupuncture effect (group I ve
group II P« .001) . Next, the results show that in group III naloxone blocks
acupuncture analgesia (group I vs group III P « ,001), while the saline control
group IV has no such effect (group I vs group IV P = 0 .25) . Consequently group
IV showed far more analgesia than group III (group III versus group IV P «,001) .
All the three control experiments in figure 2 showed no analgesia (no in-
creased latency) . The small decrease of latency in group VII at 60 minutes
(P<0 .05) may reflect sensitization of the nose with repeated noxious stimuli.
The larger decrease in latency observed at 40 minutes with âaloxone group V
was significantly greater than saline group VI (group V va group VI P«,001) or
no treatment group VII (group V va group VII P<,005) . This will be discussed
below in relation to hyperalgesia . Finally it should be noted that naloxone
plus acupuncture was not different to the plain naloxone treatment (group III
versus group V P>0 .7) .
DISCUSSION
Our results suggest two main conclusions . First, acupuncture may have
released endorphin which was blocked by naloxone (group III) . We are now test-
ing this conclusion by extracting endorphin during acupuncture and assaying for
endorphin on guinea pig ilium (10) . Secondly, we may conclude that the brain
releases small amounts of endorphin even without acupuncture since naloxone in-
creased pain sensitivity in groups III and V; this is consistent with findings
of endorphin in brains of normal animals (10, 11) . Naloxone hyperalgesia in
mice was previously reported (9) and dose response curves reached plateau at
about 0 .9 mg/kg .
transmission in spinal cord pain pathways (16) . Third, this brainstem analge-
sic effect is abolished by naloxone (5) .
Acknowledgements
The authors are grateful to Mary Adams, Richard Cheng and Elliot Goldner
for their assistance . This research was supported by a grant from Ontario
Ministry of Health ACB.
References