Inflammation ( # 2014)

DOI: 10.1007/s10753-014-0015-y

Attenuation of Morphine Analgesic Tolerance by Rosuvastatin

in Nave and Morphine Tolerance Rats
Yongle Li,1 Yinyin Shu,1 Qing Ji,1 Jian Liu,1 Xiaoyun He,1 and Weiyan Li1,2

AbstractRecent studies suggested that statins have anti-inflammatory effects beyond their lipidlowering properties. Since inflammation in the central nervous system was highly related to morphine
tolerance, we sought to investigate whether statins could affect morphine tolerance by mediating gliaderived proinflammatory cytokines secretion. We have undertaken two separate studies: Firstly, we
determined the effect of rosuvastatin on nave rats during induction of morphine tolerance. Secondly, we
investigated whether rosuvastation could attenuate the morphine analgesic tolerance in rats that the
morphine tolerance established previously. Results demonstrated that peroral rosuvastatin not only
delays, but also partially reverses the tolerance to morphine analgesia in rats. The administration of
rosuvastatin during induction of morphine tolerance attenuated the activation of ERK and the release of
proinflammatory cytokines in the lumbar spinal cord. Similar outcomes were observed in rats were
morphine tolerance was established previously. Moreover, our study also found that repeated administration of morphine could activate the astrocytes in the spinal cord while rosuvastation succeeds in
suppressing the activation of astrocytes. Our results support the idea that targeting glia-derived proinflammatory effects during morphine treatment is a novel and clinically promising method for enhancing
analgesic effects of morphine. We identify a potential new application of statins in the treatment of
morphine analgesic tolerance.
KEY WORDS: morphine tolerance; central inflammation; glia; cytokines; ERK.

INTRODUCTION
Morphine is a class of the most effective analgesics
for treating many forms of pain, especially for chronic
pain, such as neuropathic pain and cancer pain. However,
its effectiveness in the clinical setting has gradually diminished due to repeated administration.
Glia in the spinal cord are now known to contribute to
morphine tolerance [1, 2]. Astrocytes and microglia respond to repeated morphine in a proinflammatory fashion,
with upregulated activation markers and proinflammatory
cytokines [35]. Preemptive and repeated administration of
glial inhibitors [68], or blocking glial proinflammatory
mediators signalings, [912] can remarkably attenuate
morphine tolerance. These findings suggest that the suppression of glial activation by inhibiting the release of

Department of Anesthesiology, Jinling Hospital, School of Medicine,

Nanjing University, Nanjing, Peoples Republic of China
2
To whom correspondence should be addressed at Department of
Anesthesiology, Jinling Hospital, School of Medicine, Nanjing University,
Nanjing, People's Ripublic of China.. E-mail: liweiyannj@126.com

proinflammatory cytokine shows great promise for improving the efficacy of morphine.
Statins, known as 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, are commonly
used as cholesterol-lowering drugs. Recent evidences suggest that statins exert pleiotropic effects besides lowering
cholesterol [1316], and a significant proportion of these
effects may be considered as anti-inflammatory [17, 18]. It
has been reported that simvastatin-attenuated microglial
cells and astrocyte activation and decreased IL-1 level
after traumatic brain injury [19], and lovastatin inhibited
the induction of cytokines in rat primary astrocytes and
microglia [20]. More recent studies have demonstrated that
rosuvastatins alleviated experimental nerve injury-induced
neuropathic pain through attenuates microglial cells and
astrocytes activation and decreases IL-1 level [21]. All
these findings indicated that statins could regulate the inflammation in the central nervous system. Since morphine
analgesic tolerance was closely related to the inflammation
induced by activated glia cells in the central nervous system,
we hypothesized that rosuvastatin could interfere with the
tolerance to morphine analgesia.

0360-3997/14/0000-0001/0 # 2014 Springer Science+Business Media New York

Yongle, Yinyin, Qing, Jian, Xiaoyun, and Weiyan

In the present study, two separate experiments were
conducted to address whether rosuvastatin could delay the
development of morphine analgesic tolerance in nave rats
and attenuate the analgesic tolerance to morphine when the
tolerance in rats was previously established.

MATERIALS AND METHODS

Animals
Male SpragueDawley rats (Animal Center of Jinling
Hospital, Nanjing, China) weighing 200250 g were used
for the present study. Animals were housed in cages with
sawdust bedding under a standard 12/12 h light/dark cycle
with food and water available ad libitum. The animal use
and care protocols, including all experimental procedures,
were approved by the Southern Medical University Animal
Care and Use Committee and in accordance with the Guide
for the Care and Use of Laboratory Animals by the
National Institute of Health (NIH).

received oral gavage of either saline or rosuvastatin (0.4, 2,

or 10 mg/kg) once daily at 0830 hours, 30 min before
subcutaneous injection of either saline or morphine accordingly. Behavioral test was recorded on day 1 and 7, and
drug administration was conducted during day 2 to 6.
Lumbar spinal cord (5 mm, L4L6) tissue was harvested
after behavioral test immediately on day 7.
To investigate the effect of rosuvastatin on morphine
analgesic tolerance in rats with morphine tolerance, six
groups (n=8) of rats separately received subcutaneous
injection of either saline or morphine for 5 days to induce
morphine tolerance first. Then, on day 6, the oral gavage of
either saline or rosuvastatin (0.4, 2, or 10 mg/kg) was
administrated once daily at 0830 hours, 30 min before
subcutaneous injection of either saline or morphine.
Behavioral test was recorded on day 6 and 11, and drug
administration was conducted during day 6 to 10. Lumbar
spinal cord tissue was harvested after behavioral test immediately on day 11.
Western Blot Analysis

Tolerance to morphine analgesia was induced by subcutaneous injection of morphine (10 mg/kg) twice a day at
0900 and 1600 hours for 5 days.
To test for analgesic effect of morphine, paw withdrawal latency (s) to thermal stimulation was assessed via a
Paw Thermal Stimulator with radiant heat source focused
on the plantar surface of the right hind paw. Paw withdrawal latency was measured with a timer that was started at the
beginning of heat exposure and tripped when the paw was
removed from the heat. A cutoff time of 20 s was set to
avoid tissue damage. Three trials were made for each rat
with an intertrial interval of 5 min, and mean of three
measurements was taken as the final latency. Rats were
acclimatized to the testing chambers for at least 30 min
prior to each testing session. Nociceptive tests were performed both before and 30 min after morphine administration. The percentage of maximal possible antinociceptive
effect (%MPE) was calculated by comparing the latency
before (basic paw withdrawal latency (PWL)) and after
(Test PWL) drug injection using the following equation:
%MPE=[(Test PWLBasic PWL)/(cutoff time Basic
PWL)]100.

Lumbar spinal cord tissues were harvested from four

rats per group. Tissue samples were homogenized in RIPA
buffer and cleared by centrifugation (10,000g for 10 min).
The protein concentration in the supernatant was determined using the BCA Protein Assay Kit (Sigma).
Samples containing 50 g of protein were heated for
5 min at 100 C in loading buffer (50 mM TrisHCl, 2 %
SDS, 2 % -mercaptoethanol, 8 % glycerol, and 0.1 %
bromophenol blue) and resolved by SDS-PAGE on 10 %
polyacrylamide gels. After gel electrophoresis, proteins
were electrophoretically transferred to PVDF membranes
(Milipore). The membranes were blocked using 5 % nonfat dry milk in (Tris-buffered saline (TBST) with 0.1 %
Tween 20) for 2 h and incubated overnight at 4 C with a
primary antibody to ERK (1:1,000, Cell Signaling
Technology), p-ERK (1:1,000, Cell Signaling
Technology), and GAPDH (1:1,000, Cell Signaling
Technology). Membranes were then incubated in HRPconjugated secondary antibody (1:1,000; Cell Signaling
Technology) for 1 h at room temperature, visualized with
Super Signal West Femto Maximum Sensitivity Substrate
(Pierce) for 5 min, and exposed onto X-films for 15 min.
The X-films were scanned and the density of target bands
was analyzed by UN-SCAN-IT.

Experimental Design

ELISA

To investigate the effect of rosuvastatin on morphine

analgesic tolerance in nave rats, six groups (n=8) of rats

Lumbar spinal cord tissues were harvested from four

rats per group. Tissue samples were homogenized and

Induction of Morphine Tolerance and Behavioral Test

Rosuvastatin Attenuate Morphine Analgesic Tolerance in Rats

cleared by centrifugation. After centrifugation, supernatants were collected and stored at 80 C. The amounts
of IL-1 (R & D Systems) or TNF- (R & D Systems)
were assayed using commercially available rat-specific
ELISA kits according to the manufacturers instructions.
Immunohistochemistry
Animals were perfused transcardially with 0.9 % saline followed by 4 % paraformaldehyde in 0.1 M phosphate buffer (pH 7.4). Lumbar spinal cords (L46) were
removed and kept in the same fixative for 4 h at 4 C and
then replaced with 30 % sucrose overnight. Transverse 35m-thick sections were cut on a cryostat and processed for
immunohistochemistry. Sections were blocked with 5 %
BAS for 2 h at room temperature and then incubated
overnight at 4 C with primary antibody to GFAP
(1:1,000, Milipore). The FITC-conjugated goat secondary
antibodies were applied for 2 h at 4 C. The stained
sections were examined with a Leica SP2 confocal laserscanning microscope, and images were captured with a
CCD spot camera.
Statistical Analysis
All data were expressed as meansSEM, and significant differences between groups were compared with oneway ANOVA and followed by Bonferroni post-hoc comparison test. Statistical analyses were performed with SPSS
18.0 (SPSS Inc) with the significance level set at p<0.05.

RESULTS
Effect of Repeated Morphine Administration on Basic
Paw Withdrawal Latency
After continuous injection of morphine for 5 days, the
mean paw-withdrawal latency to thermal stimuli showed
no significant difference among the six groups on day 7
(Table 1). Then, the injection was continued for five more
consecutive days to 10 days in total, and no thermal
hyperalgesia was observed in rats whether they received
repeated morphine administration or not (Table 1).
Preemptive Peroral Rosuvastatin Delayed
the Development of Tolerance to Analgesic Effects
of Morphine
Rats that received subcutaneous injection of either
saline or morphine showed a strong analgesic effect to
the first administration of morphine on day 1, and no

significant difference was observed in the behavioral tests

(Table 2). The analgesic effect of morphine was significantly reduced on day 7 (17.71 %3.96), as compared to
the effect of morphine observed in the saline-treated rats
(82.47 %6.42). When administrated alone, rosuvastatin
(10 mg/kg, once daily, p.o.) treatment had no significant
effect on both paw withdrawal latency and MPE(%) in
saline-treated rats (Tables 1 and 2). However, preemptive
rosuvastatin treatment (2 or 10 mg/kg, once daily, p.o.)
during the induction of morphine tolerance strongly influenced the development of tolerance to morphine analgesia,
which was measured by paw withdrawal latency in rats
30 min after morphine administration (Table 2).
Rosuvastatin Partially Reverses the Established
Tolerance to Morphine Analgesia
Rats, which received subcutaneous injection of morphine for the first 5 days, showed a significant reduction in
morphine analgesic effect on day 6, as compared with rats
that received saline treatment (Table 2). On day 11, the
analgesic effect of morphine still maintains unsatisfied in
rats that only received morphine injection (13.31 %5.61).
However, after the rosuvastatin treatment (2 or 10 mg/kg,
once daily, p.o.) initiated on the sixth day and continued for
the next five consecutive days, morphine produced significant analgesia which was reflected by the MPE(%) in both
groups (48.014.69 and 59.096.95, respectively) higher
than that in group treated with morphine alone (13.31
5.61) (Table 2). In both experiments, the effect of 0.4 mg/
kg rosuvastatin treatment was too weak to reverse or even
delay the tolerance to morphine analgesia.
Effect of Rosuvastatin on ERK Activation in the Spinal
Cord in Nave Rats Tolerance to Morphine
Western blot analysis was used to measure the expression levels of ERK and p-ERK in spinal cords in nave
rats on day 7 following repeated morphine administration.
As shown in the panel of Fig. 1a, the GAPDH level is used
as loading control, and no significant difference among
groups was found in the expression of total ERK protein
after repeated morphine treatment twice daily for five
consecutive days. Compared to the controls (saline-treated
rats, S+S), repeated morphine treatment significantly enhanced expression of p-ERK, and indicated that ERK was
active in the spinal cord due to chronic morphine treatment.
However, this effect was inhibited when 2 or 10 mg/kg
rosuvastatin was co-administrated and no significant
changes in the expression of p-ERK were detected in rats

Yongle, Yinyin, Qing, Jian, Xiaoyun, and Weiyan

Table 1. Repeated Morphine Administration Did Not Induce Thermal Hyperalgesia
Treatment

Saline+saline
Saline+morphine
Rosuvastatin+saline (10)
Rosuvastatin+morphine (0.4)
Rosuvastatin+morphine (2)
Rosuvastatin+morphine (10)

8
8
8
8
8
8

Nave

Morphine tolerance

Day 1

Day 7

Day 6

Day 11

9.521.86 s
9.420.95 s
9.481.22 s
9.551.46 s
9.811.80s
9.451.32 s

9.441.86 s
9.360.95 s
9.411.23 s
9.471.46 s
9.741.80s
9.380.67 s

9.441.86 s
9.360.95 s
9.431.23 s
9.631.11 s
10.100.90s
9.541.25 s

9.011.02 s
9.391.63 s
9.300.87 s
9.341.24 s
9.400.58 s
9.061.73 s

Data are expressed as MeanSEM. Intergroup differences were analyzed by ANOVA Bonferronis Multiple Comparison Test
P<0.05 vs. Saline-Treated Rats, P<0.05 vs. Morphine-Treated Rats

treated with rosuvastatin alone (R+S) as compared to

saline-treated rats.
Effect of Rosuvastatin on ERK Activation in the Spinal
Cord in Rats with Morphine Analgesic Tolerance
As shown above, morphine analgesic tolerance had
successfully developed in rats that received subcutaneous
injection of morphine for the first 5 days (S+M, R+M),
and activation of ERK in these rats were detected by
Western blot. Similarly, on day 11, the expression of total
ERK protein had no significant difference while the expression of p-ERK was significantly increased after morphine administration lasted for ten consecutive days as
compared to the controls (saline-treated rats, S+S), indicating that ERK was still active in the spinal cord.
However, repeated rosuvastatin treatment (2 or 10 mg/kg,
p.o. for 5 days, once daily) started from day 6 significantly
decreased the level of p-ERK in the spinal cord measured
by Western blot on day 11. No significant changes in the
expression of p-ERK were detected in rats treated
with rosuvastatin alone (R+S) as compared to salinetreated rats.

Effect of Rosuvastatin on IL-1 and TNF- Release

in the Spinal Cord in Morphine Analgesic Tolerance
Repeated administration of morphine for 5 days significantly increased the release of IL-1 and TNF- at L5
lumbar spinal cord compared to saline-treated rats
(Table 3). Rosuvastatin treatment (10 mg/kg, p.o. for
5 days, once daily) had no significant effect on the release
of IL-1 and TNF- in saline-treated rats. However,
rosuvastatin treatment (2 or 10 mg/kg, p.o. for 5 days, once
daily) significantly attenuated morphine-induced upregulation of IL-1 and TNF-, no matter when during the
induction of morphine tolerance or the tolerance to morphine was well established.
Effect of Rosuvastatin on Astrocytes Activation
in Spinal Cord in Rats with Morphine Analgesic
Tolerance
Immunohistochemistry was used to investigate the
effect of rosuvastatin on the expression level of GFAP, a
specific marker of astrocyte, in spinal cords when the
tolerance to morphine analgesia in rats was established.

Table 2. Effect of Rosuvastatin on Morphine Analgesic Tolerance in Rats

Treatment

Saline+saline
Saline+morphine
Rosuvastatin+saline (10)
Rosuvastatin+morphine (0.4)
Rosuvastatin+morphine (2)
Rosuvastatin+morphine (10)

8
8
8
8
8
8

Nave (%)

Morphine tolerance (%)

Day 1

Day 7

Day 6

Day 11

88.676.50
86.755.15
87.696.10
89.5410.66
88.408.68
87.995.89

82.476.42
17.713.96*
88.595.65#
23.097.05*
55.966.84#
64.966.66#

83.717.13
21.244.77*
80.356.23
24.698.92*
22.815.54*
19.875.34*

89.507.28
13.315.61*
88.985.42#
16.765.65*
48.014.69#
59.096.95#

Data are expressed as MeanSEM. Intergroup differences were analyzed by ANOVA Bonferronis Multiple Comparison Test
*P<0.05 vs. Saline-Treated Rats, # P<0.05 vs. Morphine-Treated Rats

Rosuvastatin Attenuate Morphine Analgesic Tolerance in Rats

As shown in the panel of Fig. 2, when administered alone,
rosuvastation had no effect on the astrocytes activity measured by the expression of GFAP compared to the salinetreated rats (S+S). Furthermore, consistent with former
studies, the expression level of GFAP significantly increased after injection of morphine for 10 consecutive
days, indicating astrocytes were stimulated. However,
when co-administrated with 10 mg/kg rosuvastatin,
morphine-induced the upregulation of GFAP was significantly attenuated.

DISCUSSION

Fig. 1. Chronic morphine-induced ERK activation in spinal cord is

inhibited by co-administration of rosuvastatin. a Rosuvastation attenuate
ERK activation during acquisition of morphine tolerance in nave rats. b
Rosuvastation reverse the ERK activation in rats with morphine tolerance.
GAPDH level was used as loading controls. The results are presented as a
percentage of control (saline-treated rats, S+S) of the densitometry analysis of all samples. Values are shown as the meanSEM. Intergroup
differences were analyzed by ANOVA Bonferronis Multiple Comparison
Test (*P<0.05 vs. S+S, #P<0.05 vs. S+M).

Recently, rosuvastatin, known as a common

cholesterol-lowering drug, has been reported to play an
important role in alleviating neuropathic pain and
inhibiting inflammation induced by activated glial cells in
the spinal cord [21], whether rosuvastatin has an effect on
morphine antinociceptive tolerance is still unknown. In the
present study, consistent with Christensen et al. [22], there
was no thermal hyperalgesia in rats after repeated morphine administration. We demonstrated that peroral
rosuvastatin significantly attenuated the development of
morphine antinociceptive tolerance, and it was still effective when morphine antinociceptive tolerance had been
previously established. In parallel to the impact on the
analgesic effect of morphine, proinflammatory cytokine
release in the spinal cord was reduced by rosuvastatin
treatment. Additionally, systemic rosuvastatin treatment
significantly suppresses the ERK activation either in
nave rats or in morphine tolerance rats and attenuates the
chronic morphine-induced astrocyte activation.
Contrary to the traditional views on glial role in the
central nervous system, compelling evidences have shown
that glia, including microglia and astrocyte, were involved

Table 3. Effect of Rosuvastatin on IL-1 and TNF- Release in Spinal Cord in Rats with Morphine Analgesic Tolerance
Treatment

Saline+saline
Saline+morphine
Rosuvastatin+saline (10)
Rosuvastatin+morphine (0.4)
Rosuvastatin+morphine (2)
Rosuvastatin+morphine (10)

4
4
4
4
4
4

Nave (pg/mg)

Morphine tolerance (pg/mg)

IL-1

TNF-

IL-1

TNF-

27.980.86
73.823.93*
34.371.93#
70.410.77*
38.902.74#
31.701.92#

17.731.71
50.751.78*
18.911.21#
48.062.94*
24.611.94#
19.231.04#

26.721.99
89.064.62*
36.330.48#
96.203.39*
48.960.88#
35.602.27#

18.021.07
55.785.39*
19.371.59#
61.904.59*
30.753.18#
21.121.79#

Values are pg/mg Total ProteinSEM. Intergroup differences were analyzed by ANOVA Bonferronis Multiple Comparison Test
*P<0.05 vs. Saline-Treated Rats, # P<0.05 vs. Morphine-Treated Rats

Yongle, Yinyin, Qing, Jian, Xiaoyun, and Weiyan

Fig. 2. Chronic morphine-induced astrocytes activation in spinal cord is inhibited by co-administration of rosuvastatin. The expression of GFAP in lumbar
spinal cord were limited in rats treated with only saline and 10 mg/kg rosuvastatin (S+S, R+S) while morphine tolerance group (S+M) were increased
significantly. Ten milligrams per kilogram rosuvastatin treatment (R10+M) downregulated the enhanced expression of GFAP in morphine tolerance rats.

in creating and maintaining morphine tolerance states [2,

2325]. Inhibition of glial activation by glial inhibitors
such as fluorocitrate [1] or propentofylline [6] significantly
blocked morphine antinociceptive tolerance. Similarly, intrathecal pretreatment with minocycline [7], a selective
microglia activation inhibitor, attenuates the development
of morphine anticiceptive tolerance as well as the activities
of the spinal microglia and astrocyte induced by chronic
morphine treatment. But interestingly, intrathecal
minocycline administration fails to reverse the established
morphine anticiceptive tolerance which correlated with the
observation that only the activation of microglia, but not
astrocytes is inhibited. It seems to suggest that the spinal
microglia activation might be mainly associated with the
initiation of morphine analgesic tolerance, while the astrocytes activity is with the maintenance of tolerance. This
hypothesis was further supported by the latest findings
[26], in which, IL-18, members of IL-1 family, released from
the mircoglia led to the activation of astrocytes. In the present
study, rosuvastatin partially reverses the established
anticiceptive tolerance to morphine. It has been demonstrated
that rosuvastatin could suppress the activation of both the
spinal microglia and astrocytes [21, 27]. As shown in our
results, reversal of chronic morphine treatment-induced astrocyte activation by co-administration of rosuvastatin was
observed in immunohistochemistry, indicating that inhibition

of astrocyte activation might contribute to the additional

effect of rosuvastatin on established morphine analgesic
tolerance.
As a consequence of glia activation, proinflammatory
cytokines were released and the interaction between morphine tolerance and cytokines has been well explored [28
30]. In early studies, Raffa et al. [31] first postulated that
cytokines might interact with the opioid receptor and modulate its actions and Gul et al. [32] observed a reduction in
the analgesic effect of morphine after exogenous administration of IL-1. Consistent with previous findings, Shavit
et al. [9] further discovered that mice with genetic or pharmacological blockade of IL-1 signaling displayed no tolerance following repeated administration of morphine and the
analgesia was reinstated when acute IL-1ra was administrated in control mice, either immediately following the
cessation of acute morphine analgesia, or following the
development of chronic morphine tolerance. These findings
suggested that the analgesic effect of morphine is unmasked
when the IL-1 system is blocked. Additionally, upregulation
of TNF- has also been observed during the development
of morphine tolerance [2, 33, 34]. A recent study showed
that etanercept, a TNF- biological antagonist, not only
preserved the antinociceptive effect of morphine in
morphine-tolerant rats, but also suppressed proinflammatory cytokines TNF-, IL-1, and IL-6 mRNA expression

Rosuvastatin Attenuate Morphine Analgesic Tolerance in Rats

[12]. In agreement with previous results, we have shown
that repeated morphine administration induced a significant
increase in IL-1 and TNF- in the spinal cord. Since the
anti-inflammatory effects of statins have been extensively
studied, we investigated the influence of rosuvastatin on
these glia-derived proinflammatory substances. Our results
demonstrated that 2 or 10 mg/kg rosuvastatin effectively
downregulated the release of IL-1 and TNF-, suggesting
the possibility that the ability of rosuvastatin to reduce
cytokine secretion in the periphery could play a role in their
ability to alleviate analgesic tolerance to morphine.
There is substantial evidence indicating that mitogenactivated protein kinase (MAPK), including extracellular
signal-regulated protein kinase (ERK), can be activated by
chronic morphine treatment in the central and peripheral
nervous systems [3539]. To further understand the molecular mechanisms underlying the role of rosuvastatin in
inhibiting tolerance to morphine analgesia, we studied the
effect of peroral rosuvastatin on ERK activation in the spinal
cord. Previous studies have shown that intrathecal injection
of morphine (15 g/day) for 7 days induced activation of
ERK in the spinal cord of rats [40] and that inhibition of
ERK activation by an ERK inhibitor U0126 [41] or
PD98059 [40] or knockdown of the spinal ERK by antisense oligonucleotides [42, 43] attenuates the tolerance to
morphine analgesia. In accordance with the previous results,
we observed a significant increase in the level of p-ERK in
rats under repeated morphine administration. Interestingly,
this effect was inhibited when 2 or 10 mg/kg rosuvastatin
was co-administrated either following the induction of morphine tolerance or the morphine tolerance was established.
According to Wang et al. [40], chronic treatment with
morphine increases p-ERK levels in the lumbar spinal cord
and p-ERK was mostly restricted locally in astrocytes.
Additionally, an interesting finding in his study indicated
that the activation of the ERK pathway in astrocytes leads to
the enhanced expression of various downstream targets
including IL-1 and TNF-. That might provide reasonable
explanations for the effects of rosuvastatin on morphine
tolerance, in that, peroral rosuvastatin attenuates morphine
analgesic tolerance by suppressing the activation of ERK
pathway, which has a close relationship with astrocyte activity as well as IL-1 and TNF- secretion.
In summary, our results show that peroral rosuvastatin
not only delayed, but also partially reverse tolerance to
morphine analgesia in rats. Furthermore, it is suggested
that the inhibition of ERK activation, which have a close
relationship with astrocyte activity as well as IL-1 and
TNF- secretion that lead to central inflammation, is one
essential mechanism thereby rosuvastatin attenuates

morphine antinociceptive tolerance. Since, there have already been accumulating human studies that have demonstrated the anti-inflammatory effects of statins on reducing
cytokine levels in patients at clinically relevant doses [44
46], it suggests that the results in our study could have
clinical consequences, in which means statins may represent a novel therapeutic strategy in the treatment of morphine tolerance.

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