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. 2013 Feb;19(2):83-90.
doi: 10.1111/cns.12035. Epub 2012 Dec 11.

Electroacupuncture suppresses mechanical allodynia and nuclear factor κ B signaling in streptozotocin-induced diabetic rats

Lei Shi  1 , Hong-Hong Zhang, Ying Xiao, Ji Hu, Guang-Yin Xu
Affiliations

Electroacupuncture suppresses mechanical allodynia and nuclear factor κ B signaling in streptozotocin-induced diabetic rats

Lei Shi et al. CNS Neurosci Ther. 2013 Feb.

Abstract

Aims: To investigate whether electroacupuncture (EA) produced analgesic effect and whether nuclear factor kappa B (NF-κB) and cystathionine β synthase (CBS) involved in EA-mediated analgesia in painful diabetic neuropathy in rats.

Methods: Diabetes was induced by an intraperitoneal injection of streptozotocin (STZ) in adult female rats. Mechanical pain threshold was measured by von Frey filaments. EA was applied at acupoint Zu-San-Li (ST-36) in both hindlimbs. Western blot analysis was employed to detect changes in protein levels of NF-κB and CBS in spinal dorsal root ganglion (DRGs).

Results: Mechanical allodynia was developed 2 weeks after STZ injection and lasted for another 4 weeks. STZ injection significantly enhanced expression of p65 and CBS in lumbar L4-6 DRGs when compared with age-matched controls. EA markedly attenuated mechanical allodynia. Importantly, EA treatment remarkably inhibited p65 and CBS expression in DRGs. Additionally, intrathecal injection of the p65 antagonist pyrrolidine dithiocarbamate attenuated mechanical allodynia and markedly inhibited CBS expression in DRGs in STZ rats.

Conclusions: These data indicate that EA produced an analgesic effect, which might be mediated at least in a part by inhibition of NF-κB signaling pathway in primary sensory neurons in rats with diabetes.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Hyperglycemia and mechanical allodynia in streptozotocin (STZ)‐induced diabetic rats. (A) Following a single i.p. injection of STZ, the fasting blood glucose level was significantly enhanced 1 week later. The enhancement persisted for at least another 11 weeks within our observation period of time. *P < 0.01 versus control, n = 6 for both groups. (B) The growth rate of STZ rats was reduced. The fasting body weight (FBW) of control rats exhibited a steady increase, whereas the FBW of STZ rats remained unchanged. *P < 0.05 versus control, n = 6 for both groups. (C) Paw withdrawal threshold (PWT) in response to von Frey filament stimulation was significantly lowered 2 weeks and reached the lowest point 4 weeks after STZ injection. The mechanical allodynia lasted for at least 4 weeks. Data points are expressed as mean ± SEM. Error bars smaller than the size of symbols are not shown. *P < 0.01 versus control, n = 6 for both groups.
Figure 2
Figure 2
NF‐κB antagonist significantly attenuates streptozotocin‐induced mechanical allodynia. (A) Effects of a single intrathecal injection of pyrrolidine dithiocarbamate (PDTC). A single injection of PDTC (1 μg) significantly increased the paw withdrawal thresholds (PWTs), while NS injection or PDTC (0.1 μg) did not show any significant change in PWTs in diabetic rats when compared with those before injection (Pre). (B) Effects of PDTC injection once every day for consecutive 7 days. Note that the antiallodynia effect of PDTC (1 μg) lasted for 3 days. *P < 0.05 versus Pre, #P < 0.05 versus NS, & P < 0.05 versus 0.1 μg group, n = 7.
Figure 3
Figure 3
Streptozotocin (STZ) treatment enhances expression of p65 and cystathionine β synthase (CBS). Proteins were extracted from L4‐6 dorsal root ganglions in rats 4 weeks after STZ or citrate buffer injection. (A) The expressions of p65 were significantly enhanced in STZ‐injected rats (n = 4) when compared with controls (CON, n = 3). **P < 0.01 compared with those of CON. (B) CBS was significantly enhanced in STZ‐injected rats (n = 5) when compared with controls (n = 5). **P < 0.01 compared with those of CON. The protein expressions were normalized with their respective β‐actin.
Figure 4
Figure 4
Pyrrolidine dithiocarbamate (PDTC) treatment reduces expression of cystathionine β synthase (CBS). The expression CBS in L4‐6 dorsal root ganglion was significantly reduced after 1‐week PDTC treatments (n = 5) in streptozotocin‐injected rats when compared with controls (NS, n = 6). The protein expressions were normalized with their respective β‐actin *P < 0.05.
Figure 5
Figure 5
Inhibitory effect of electroacupuncture (EA) on mechanical threshold. (A) Effect of one‐time EA treatment (30 min). EA at ST‐36 produced the analgesic effect in streptozotocin (STZ) rats when compared with STZ rats with sham EA treatment (sham). Sham EA at ST‐36 and EA at BL‐43 did not produce any effect in STZ rats (BL‐43). n = 7 for each group. (B) Time course of prolonged analgesic effect of EA treatment. EA treatments were given once every day (30 min) for seven consecutive days in STZ rats. EA at ST‐36 greatly enhanced the mechanical threshold in STZ‐injected rats. This effect lasted for about 5 days. The sham EA treatment had no effect on paw withdrawal threshold (PWT). n = 7 for each group. *P < 0.05 when compared with values of sham. #P < 0.05 when compared with before EA treatment (Pre). (C) One‐time EA treatment at BL‐43 did not produce any effect on PWT. (D) EA treatment for seven consecutive days in STZ rats at BL‐43 did not produce any effect on PWT.
Figure 6
Figure 6
Inhibitory effect of electroacupuncture (EA) on p65 and cystathionine β synthase (CBS) expression. EA treatments at ST‐36 once every day (30 min) for seven consecutive days significantly inhibited the expression of p65 (A, n = 6 for each group, *P < 0.05) and CBS (B, n = 8 for each group, *P < 0.05) in L4‐6 dorsal root ganglion from streptozotocin rats when compared with sham EA. The protein expressions were normalized with their respective β‐actin.
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