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.

ObjectiveTo clarify the differences in the efficacy and mechanism of different processed products of Atractylodes chinensis rhizoma by the pharmacodynamics and metabolomics studies of raw， bran-fried and rice water-processed products on rats with spleen deficiency. MethodSixty male SD rats were randomly divided into blank group， model group， raw product group（3.75 g·kg^-1）， bran-fried product group（3.75 g·kg^-1）， rice water-processed product group（3.75 g·kg^-1） and Shenling Baizhusan group（6.7 g·kg^-1）， with 10 rats in each group. The method of excessive fatigue+improper diet was used to establish a spleen deficiency model in rats. After the end of modeling， except for the blank and model groups， each dosing group was given the corresponding drug suspension， the immune organ coefficients of each group of rats were examined， the levels of interleukin-6（IL-6）， tumor necrosis factor-α（TNF-α）， immunoglobulin G（IgG）， amylase（AMS）， motilin（MTL）， gastrin（GAS）， Na⁺-K⁺-adenosine triphosphatase（ATPase）， aquaporin 2（AQP2）， AQP3 and AQP8 in rats were measured by enzyme-linked immunosorbent assay（ELISA）. Ultra high performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry（UPLC-Q-TOF-MS） combined with orthogonal partial least squares-discriminant analysis（OPLS-DA） were used to search for biomarkers in the plasma samples of spleen-deficient rats by using two criteria［P<0.05 and variable importance in the projection（VIP） value>1］， and to compare the different modulatory effects of the three decoction pieces on the splenic-deficient biomarkers， and metabolic pathway analysis was conducted through the Kyoto Encyclopedia of Genes and Genomes（KEGG） database. ResultCompared with the blank group， the thymus index and spleen index of rats in the model group were significantly decreased（P<0.05）， the levels of IL-6， TNF-α， IgG and AQP2 were significantly increased（P<0.05）， the levels of AMS， GAS， MTL， AQP3， AQP8 and Na⁺-K⁺-ATPase were significantly decreased（P<0.05）. Compared with the model group， raw products， bran-fried products and rice water-processed products all increased thymus index and spleen index（P<0.05）， decreased IL-6， TNF-α， IgG and AQP2 levels（P<0.05）， and increased AMS， GAS， MTL， AQP3， AQP8 and Na⁺-K⁺-ATPase levels to different degrees. A total of 176 differential metabolites were screened in the model group compared with the blank group， of which 75， 72 and 84 biomarkers were called back by the raw products， bran-fried products and rice water-processed products， respectively（P<0.05， P<0.01）. Raw products of A. chinensis rhizoma mainly affected glycine， serine and threonine metabolism. Bran-fried products mainly affected alanine， aspartate and glutamate metabolism， D-arginine and D-ornithine metabolism. Rice water-processed products mainly affected glycine， serine and threonine metabolism， alanine， aspartate and glutamate metabolism， citrate cycle， thiamine metabolism， D-arginine and D-ornithine metabolism. ConclusionRaw products， bran-fried products and rice water-processed products of A. chinensis rhizoma all have good spleen strengthening effects， among which the effects of bran-fried products and rice water-processed products were stronger. Meanwhile， raw products has the strongest dryness， followed by bran-fried products， and the weakest dryness of rice water-processed products. The three decoction pieces are able to significantly modulate metabolic abnormalities in spleen-deficient rats， and the mechanism may be related to amino acid metabolism such as glycine， serine and threonine metabolism as well as alanine， aspartate and glutamate metabolism.