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Preventing the transition from acute to chronic pain (pain transition) is a new strategy for treating chronic pain. The present study aimed to investigate the role of K+-Cl- Cotransporter Isoform 2 (KCC2) and γ-aminobutyric acid receptor type A (GABAAR) in the spinal cord dorsal horn (SCDH) in pain transition and the intervention effect of electroacupuncture (EA), and to understand the mechanism of EA in preventing acute and chronic pain transition in the spinal center.
A rat model of hyperalgesic priming (HP) was established by injecting carrageenan (Car) into the plantar area of rats, followed by the injection of prostaglandin E2 (PGE2) into the dorsal foot 7 days later. The GABAAR agonist (muscimol) and KCC2 activator (CLP257) were intrathecally injected for three consecutive days after PGE2 injection. EA was applied at a frequency of 2/100 Hz to the bilateral foot Zusanli (ST36) and Kunlun (BL60). A von Frey filament was used to detect the pain threshold in each group of rats. Western blotting (WB) and immunofluorescence (IF) were used to detect GABAAR and KCC2 expression in each rats group. By combining EA intervention with a KCC2 inhibitor (VU0240551), we explored the mechanism of pain transition of EA regulation of GABAAR and KCC2 expression in SCDH.
The HP model was established by injecting mice with Car/PGE2. Compared to the normal saline (NS) + NS and NS + PGE2 groups, the pain threshold of the Car + PGE2 group decreased significantly 48 hours after PGE2 injection (P < 0.01). The WB results indicated that intrathecal injection of a GABAAR agonist upregulated GABAAR expression in the SCDH of HP model rats (P < 0.05). WB and IF results revealed that intrathecal injection of the KCC2 activator significantly increased GABAAR and KCC2 expression in the SCDH of HP model rats (P < 0.01) and that GABAAR and KCC2 were co-expressed in the same SCDH cells. Compared to the Car + PGE2 group, EA intervention significantly increased MWTs from 48 to 72 hours after the first injection and 4, 24, and 48 hours after the second injection (P < 0.01). EA upregulated GABAAR and KCC2 expression in the SCDH of rats with HP (P < 0.05). Intrathecal injection of the KCC2 inhibitor blocked the analgesic effect of EA in HP model rats (P < 0.01).
In SCDH, KCC2 expression was downregulated, causing downregulation of GABAAR expression and resulting in pain transition. EA upregulates KCC2 and GABAAR expression and prevents pain transition.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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