Background
Methods
Animals and treatments
Intrathecal implantation of catheter
STZ treatment
Tactile allodynia
Cold allodynia
Tail-flick test
Spinal cord preparation
Quantitative autoradiography
Confocal microscopy
Quantification of Iba-1 immunolabelling
SYBR green-based quantitative RT-PCR
Sequences | Position | Gen Bank | |||||
---|---|---|---|---|---|---|---|
18S forward | 5' | TCA ACT TTC GAT GGT AGT CGC CGT | 3' | 363 | - | 386 | X01117 |
18S reverse | 5' | TCC TTG GAT GTG GTA GCC GTT TCT | 3' | 470 | - | 447 | |
B1 receptor forward | 5' | GCA GCG CTT AAC CAT AGC GGA AAT | 3' | 367 | - | 391 | NM_030851 |
B1 receptor reverse | 5' | CCA GTT GAA ACG GTT CCC GAT GTT | 3' | 478 | - | 454 | |
IL-1β forward | 5' | TGT CAC TCA TTG TGG CTG TGG AGA | 3' | 247 | - | 270 | NM_031512 |
IL-1β reverse | 5' | TGG GAA CAT CAC ACA CTA GCA GGT | 3' | 411 | - | 388 | |
TNF-α forward | 5' | ATG ATC CGA GAT GTG GAA CTG GCA | 3' | 160 | - | 183 | NM_012675 |
TNF-α reverse | 5' | AAT GAG AAG AGG CTG AGG CAC AGA | 3' | 257 | - | 234 | |
TRPV1 forward | 5' | GCA CAA TGG GCA GAA TGA CAC CAT | 3' | 575 | - | 598 | NM_031982 |
TRPV1 reverse | 5' | GGC ATT GAC AAA CTG CTT CAG GCT | 3' | 656 | - | 633 |
Drugs
Statistical analysis
Results
Influence of microglia inhibitors and B1R antagonists on STZ-induced hyperglycemia
Effect of microglia inhibitors on Iba-1 microglial immunoreactivity
Effect of microglia inhibitors on mRNA levels of B1R, TRPV1, IL-1β and TNF-α in STZ thoracic spinal cord
gene mRNA/18S mRNA (fold change) | ||||
---|---|---|---|---|
B1R | TRPV1 | IL-1β | TNF-α | |
Control | 1.0 ± 0.4 | 1.0 ± 0.1 | 1.0 ± 0.6 | 1.0 ± 0.6 |
STZ | 27.3 ± 8.7 * | 3.1 ± 0.4 * | 77.9 ± 19.6 * | 6.0 ± 2.1 ** |
STZ + Fluorocitrate | 2.7 ± 1.1 | 2.3 ± 0.3 + | 10.1 ± 7.9 | 0.9 ± 0.4 + |
STZ + Minocycline | 3.0 ± 1.3 | 2.6 ± 0.2 + | 8.9 ± 4.2 | 2.1 ± 0.7 + |
Effect of fluorocitrate on B1R specific binding sites in STZ thoracic spinal cord
Effect of microglia inhibitors on thermal hyperalgesia mediated by des-Arg9-BK
Effect of microglia inhibitors and B1R antagonists on allodynia induced by STZ
Effect of microglia inhibitors on enhanced tactile allodynia induced by des-Arg9-BK in STZ-treated rats
Tactile allodynia (g) | ||||||
---|---|---|---|---|---|---|
0 min | 5 min | 10 min | 15 min | 30 min | 60 min | |
Control rats
| ||||||
Vehicle | 9.7 ± 0.3 | 9.7 ± 0.3 | 9.3 ± 0.7 | 9.7 ± 0.3 | 9.7 ± 0.3 | 9.7 ± 0.3 |
DABK (9.6 nmol, i.t.) | 9.7 ± 0.3 | 9.7 ± 0.3 | 9.3 ± 0.7 | 9.7 ± 0.3 | 9.7 ± 0.3 | 9.7 ± 0.3 |
SSR240612 (10 μg, i.t.) + DABK | 10.7 ± 0.3 | 9.7 ± 0.3 | 9.3 ± 0.7 | 9.7 ± 0.3 | 9.7 ± 0.3 | 9.7 ± 1.3 |
FLU (1 nmol, i.t.) + DABK | 9.7 ± 0.3 | 9.7 ± 0.3 | 9.3 ± 0.7 | 9.7 ± 0.3 | 9.7 ± 0.3 | 9.7 ± 0.3 |
Mino (10 mg/kg, i.p.) + DABK | 9.7 ± 0.3 | 9.7 ± 0.3 | 9.3 ± 0.7 | 9.7 ± 0.3 | 9.7 ± 0.3 | 9.7 ± 0.3 |
Tactile allodynia (g)
| ||||||
0 min | 5 min | 10 min | 15 min | 30 min | 60 min | |
STZ-treated rats
| ||||||
Vehicle | 6.3 ± 0.3*** | 6.0 ± 0.5*** | 6.3 ± 0.3*** | 6.3 ± 0.3*** | 6.3 ± 0.3*** | 6.0 ± 0.5*** |
DABK (9.6 nmol, i.t.) | 6.0 ± 0.5*** | 5.3 ± 0.4*** | 4.3 ± 0.3++ *** | 5.3 ± 0.4*** | 6.0 ± 0.0*** | 5.7 ± 0.3*** |
SSR240612 (10 μg, i.t.) + DABK | 9.3 ± 0.3+++ | 9.3 ± 0.0+++ | 9.3 ± 0.0+++ | 9.3 ± 0.3+++ | 9.7 ± 0.3+++ | 9.3 ± 0.0+++ |
FLU (1 nmol, i.t.) + DABK | 9.7 ± 0.3+++ | 9.7 ± 0.3+++ | 9.7 ± 0.3+++ | 9.7 ± 0.3+++ | 9.3 ± 0.4+++ | 9.3 ± 0.4+++ |
Mino (10 mg/kg, i.p.) + DABK | 9.7 ± 0.3+++ | 9.0 ± 0.4+++ | 9.3 ± 0.4+++ | 9.3 ± 0.4+++ | 8.7 ± 0.4+++ | 9.3 ± 0.4+++ |