Effects of Guhong injection on ATPase activity and inflammatory response after cerebral ischemia/reperfusion injury in rats / 中国中西医结合急救杂志

ABSTRACT

Objective To investigate the influence of Guhong injection (GHI) on ATPase activity and inflammatory response after cerebral ischemia/reperfusion (I/R) injury in rats, and evaluate its protective effects on cerebral I/R injury. Methods Seventy-two male Sprague-Dawley (SD) rats were divided into sham group, I/R group, nimodipine group, and the low-dose (2.5 mL/kg, GHI-L), medium-dose (5.0 mL/kg, GHI-M), and high-dose (10.0 mL/kg, GHI-H) of GHI groups according to the random number table method, with 12 rats in each group. The middle cerebral artery occlusion (MCAO) model was established by the intraluminal suture method to prepare the model of focal cerebral ischemia, and reperfusion was performed after 1.5 hours of occluding the middle cerebral artery; the sham group had the same operation process except inserting the nylon thread. The injection of drug in various drug-treated groups was carried out via a tail vein at 0, 12, 24 hours after the onset of reperfusion, while the sham group and I/R group received the same amount of normal saline. At 12 hours after last drug administration, the scores of neurological deficit symptoms were evaluated; the cerebral infarction was observed by triphenyl tetrazolium chloride (TTC) staining; the Na+-K+-ATPase and Ca2+-ATPase activities in the brain tissue were measured by phosphorus determination; the contents of interleukin-6 (IL-6), monocyte chemotactic factor-1 (MCP-1), nitric oxide (NO) in serum were detected by enzyme linked immunosorbent assay (ELISA). Results Compared with the sham group, the neurological function score was significantly decreased, the cerebral infarction was serious, the activities of ATPase was obviously decreased, and the levels of serum inflammatory factors were significantly increased in I/R group. Compared with the I/R group, the neurological function scores were significantly increased in GHI-L group, GHI-M group, GHI-H group and nimodipine group (9.03±0.63, 10.54±2.55, 12.33±1.87, 12.06±1.89 vs. 8.17±1.05, all P < 0.05), the volumes of cerebral infarction were obviously reduced [(18.51±1.80)%, (15.98±1.34)%, (8.61±1.16)%, (8.09±0.96)% vs. (26.52±2.07)%, all P < 0.01], the activities of ATPase were significantly increased [Na+-K+-ATPase (μmol·mg-1·h-1):5.10±0.30, 5.34±0.26, 6.19±0.17, 5.86±0.31 vs. 3.98±0.35, Ca2+-ATPase (μmol·mg-1·h-1): 3.68±0.44, 4.43±0.29, 5.03±0.27, 4.17±0.30 vs. 1.87±0.46, all P < 0.01], and the levels of serum inflammatory factors were decreased obviously [IL-6 (ng/L): 51.61±5.55, 43.88±4.05, 39.71±2.22, 41.28±2.66 vs. 60.11±6.61, MCP-1 (ng/L): 227.82±7.07, 201.58±13.10, 177.23±10.46, 126.80±8.49 vs. 296.01±12.85, NO (μmol/L): 54.48±3.23, 46.84±2.69, 41.15±2.80, 48.62±2.34 vs. 65.25±3.88, all P < 0.05]. Conclusions GHI not only can improve the energy metabolism of brain tissue in a dose-dependent manner, but also inhibit the inflammatory cascade of damage after cerebral I/R in rats, which might be its protective mechanism on cerebral ischemia injury.