ABSTRACT
ObjectiveTo observe the effect of Huangqi Baihe granules on the hypoxia-inducible factor 1α （HIF-1α）/nuclear factor-κB （NF-κB）/NOD-like receptor hot protein domain related protein 3 （NLRP3） signaling pathway in a rat model of high altitude hypoxia. MethodSixty male SPF SD rats were randomly divided into blank group， model group， dexamethasone group （5 mg·kg-1）， and high， middle， and low-dose groups of Huangqi Baihe granules （4.1， 2.05， 1.025 g·kg-1）. Among them， each Chinese medicine group was administrated orally for continuously 14 d， once a day， and the dexamethasone group was injected intraperitoneally for continuously 3 d as the positive control group. On the 15th d， the model group， dexamethasone group， and high， middle， and low dose groups of Huangqi Baihe granules were exposed to the simulated high altitude， low pressure， and low oxygen environment in the animal low-pressure simulation cabin， and the exposure lasted for 3 d. Blood was collected from the abdominal aorta and serum was separated， and the brain tissue was taken after being killed. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in brain tissue. Enzyme-linked immunosorbent assay （ELISA） was used to detect the content of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β） in rat serum. Western blot was used to detect HIF-1α， NLRP3， phosphorylated nuclear factor-κB （p-NF-κB）， NF-κB， desquamation D （GSDMD）， and cysteine aspartate-specitis protein-1（Caspase-1） in rats of each group. The mRNA expression levels of HIF-1α， NLRP3， NF-κB p65， GSDMD， and Caspase-1 were detected by real-time quantitative polymerase chain reaction （Real-time PCR）. ResultThe results of HE staining showed that as compared with the normal group， the pathological sections of brain tissues in the model group showed that pyramidal cells were loosely arranged and distributed in disorder， with different sizes. Compared with the model group， the pathological changes in pyramidal cells in the dexamethasone group and high and middle-dose groups of Huangqi Baihe granules were reduced. The results of ELISA showed that as compared with the normal group， the content of TNF-α， IL-6， and IL-1β in the serum of rats in the model group was significantly higher （P<0.01）. Compared with the model group， the content of TNF-α， IL-6， and IL-1β in the serum of rats in the dexamethasone group and high and middle-dose groups of Huangqi Baihe granules decreased significantly （P<0.05， P<0.01）. The results of Western blot showed that as compared with the normal group， the relative protein expression levels of HIF-1α， NLRP3， p-NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of the model group were significantly higher （P<0.01）. As compared with the model group， the relative expressions of HIF-1α， NLRP3， p-NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of rats in the dexamethasone group and the high-dose group of Huangqi Baihe granules were significantly decreased （P<0.05， P<0.01）. The relative protein expression levels of HIF-1α， NLRP3， p-NF-κB p65， and Caspase-1 in the brain tissue of rats in the middle-dose group of Huangqi Baihe granules decreased significantly （P<0.01）， and the relative protein expression of HIF-1α in the brain tissue of rats in the low-dose group of Huangqi Baihe granules was reduced （P<0.05）. The Real-time PCR analysis showed that as compared with the normal group， the mRNA expression levels of HIF-1α， NLRP3， NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of the model group were significantly increased （P<0.01）. As compared with the model group， the mRNA expression levels of HIF-1α， NLRP3， NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of rats in the dexamethasone group were significantly decreased （P<0.01）. The mRNA expression levels of HIF-1α， NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of rats in the high-dose group of Huangqi Baihe granules decreased significantly （P<0.01）. The mRNA expression levels of HIF-1α， NLRP3， and Caspase-1in the brain tissue of rats in the middle-dose group of Huangqi granules decreased （P<0.05， P<0.01）. ConclusionThe protective effect of Huangqi Baihe granules on acute brain injury in low-pressure hypoxic rats may be related to the HIF-1α/NF-κB/NLRP3 signaling pathway.
ABSTRACT
Objective To determine the optimum extraction and purification technology of Huangqi Baihe Granules (HBG) by using orthogonal design and combination empowerment based on G1-entropy method, so as to provide a reference for the industrial production. Methods With the ethanol extraction amount and paste-forming rate of calycosin7-O-β-D-glucopyranoside, hesperidin, crude polysaccharide as evaluation index, and with the extraction times, extraction time and the water adding amount as investigate factors, then the combination empowerment method based on G1-entropy method and orthogonal design were used to optimize the extraction technology of HBG. The same combined methods were used to optimize the purification technology of HBG with retention rate and removal rate of impurity of ethanol extract of calycosin7-O-β-D-glucopyranoside, hesperidin and crude polysaccharide, as evaluation index, and with the membrane pore diameter, operating pressure and filtration temperature of multi-channel tubular ceramic ultrafiltration membrane as investigate factors. Results The test results showed that the optimum extraction technology was as follows: Extracted 2 times with 20 times the amount of water and each for 75 min. The optimum ultrafiltration technology was as follows: Multi-channel tubular inorganic ceramic membrane at 50 nm, operating pressure at 0.10 MPa, filtration temperature at 45 ℃. Under such condition, there was no significant difference between verification groups of three batches. Conclusion The optimized extraction and purification process is stable and feasible by verification, which can provide experimental basis for industrial production of HBG.
ABSTRACT
Objective To optimize the vacuum drying process of extract paste of Huangqi Baihe Granules (HBG) and evaluate the physical quality of powder. Methods With drying temperature, vacuum degree, and material thickness as independent variables, the comprehensive evaluating indexes of content of calycosin 7-O-β-D-glucopyranoside, hesperidin, crude polysaccharide, ethanol extraction amount, and drying rate as response values, Box-Behnken design-response surface methodology and G1-entropy method were used to optimize the vacuum drying process. The similarity of fingerprints between extract powder dried by the optimized technology and extract paste was compared. Additionally, the properties of powder were evaluated comprehensively with nine physical indicators, including relative homogeneity index, bulk density, tap density, interparticle porosity, compressibility, Hausner ratio, angle of repose, moisture content, and hygroscopicity. The physical fingerprint of powder were established to evaluate the quality consistency of different batches of extract powder. Results The optimal drying parameter was as follows: the drying temperature was 68 ℃, the vacuum degree was 0.07 MPa, the material thickness was 6 mm. Three verification experiments were carried out under these conditions and the average comprehensive evaluating indexes of vacuum drying was 91.05, which was close to the model prediction 91.87, and the relative error was 0.89%. Compared with the extract paste, the similarity of fingerprint of extract powder were more than 0.91. The similarity of chemical and physical fingerprint of three batches of extract powder were higher than 0.99. Conclusion The optimized vacuum drying technology of extract paste of HBG is stable and feasible.