ABSTRACT
This paper was aimed to study the effect of ethyl acetate extracts of Gentianella acuta on the gene and protein of insulin significant signal IRS-1 and Akt in insulin resistance (IR) HepG2 cells.The CCK-8 method was used to detect the HepG2 cell activity.HepG2 cells of human liver cancer were cultured with high concentration insulin (10-6 mol· L-1)for 36 hours to establish IR cell model.According to the results of CCK-8,the control group,model (IR) group,ethyl acetate extracts of Gentianella acuta IR + 50 μg· mL-1,IR + 500 μg· mL-1 group,and the metformin group were divided.Glucose consumption was measured with a glucose assay kit.The expressions of IRS-1 and Akt gene in IR HepG2 cells were detected by RT-PCR after 6-hour using of ethyl acetate extracts of Gentianella acuta.Western blot was used to detect the expression of IRS-1 and Akt protein after 6-hour using of ethyl acetate extracts of Gentianella acuta.The results showed that when the concentration of ethyl acetate extracts of Gentianella acuta was 500 μg· mL-1,the survival rate reached 95％.When the concentration was higher than 500 μg· mL-1,the survival rate decreased.Compared with the IR group,the IR + 50 μg· mL-1 group and the IR + 500 μg· mL-1 group promoted glucose consumption of IR HepG2 cells,but its effect was less than that of the metformin hydrochloride group.The expression of IRS-1 and Akt in IR HepG2 cells was significantly increased by using RT-PCR in the group of IR + 50 μg· mL-1 and IR + 500 μg·mL-1 compared with the IR group after 6-hour using of ethyl acetate extracts of Gentianella acuta.The expression of IRS-1 and Akt protein in the group of IR + 50 μg· mL-1 and IR + 500 μg· mL-1 was significantly higher than that in the IR group after 6-hour medication detected by western blot.It was concluded that the ethyl acetate extracts of Gentianella acuta can increase the expression of IRS-1,Akt gene,the expression of IRS-1 and Akt protein in HepG2 cells,which may be the mechanism of IR improvement.
ABSTRACT
Through comprehensively characterizing components in blood after oral administration of Tang-Bi-Kang (TBK) granules by UPLC-ESI-MSn,this study was aimed to explain the pharmaceutical material basis of TBK initially.UPLC-LTQ-Orbitrap was used under both positive and negative ion modes of electrospray ionization.The blank serum and rat serum after oral administration of TBK were analyzed.Components in rat serum were identified and characterized based on ion fragment information,evenelectron law,nitrogen rule and so on.Reference data was used to establish the UPLC-ESI-MSn method.The results showed that after oral administration of TBK granules,15 components were detected in the serum,of which 13 components were taken as the prototype to blood and 2 metabolites.It was concluded that constituents of TBK granules in rat serum were generated from compatibility of all herbal medicines.In rat serum,most of the components had been absorbed by rat's metabolism;a few were absorbed as the prototype.This research provided references for pharmacodynamic material basis and metabolism of TBL granules in vivo.
ABSTRACT
This paper was aimed to study the albiflorin activity of Tang-Bi-Kang (TBK) granules in protecting Schwann cells (SCs) of rat's sciatic nerve.The establishment of SCs oxidative stress model was the condition of 150 mmol×L-1 Dglucose with different concentrations.And the incubation time was 48 h.The experiment groups were the high-dose,middle-dose and low-dose (100 μM,20 μM,4 μM) albiflorin group,the model group,the vitamin C (100 μM) group,and the normal group.Flow cytometry was used to detect the content of ROS in SCs.Fluorescence microscope was used to observe the condition of ROS fluorescence in SCs.And CCK-8 was used to detect the cell activity.The results showed that by using CCK-8 to detect cell proliferation,after 48 h,there was a significant difference between the model group and the normal group (P＜0.01);the albiflorin group compared with the model group (P＜0.01).It indicated that albiflorin can promote the proliferation of SCs.Detecting the ROS fluorescence content,it showed that compared with the model group (Glu 150 mM),the 100 μM,20 μM,and 4 μM albiflorin group,it was P＜0.01 for each group.It showed that albiflorin could relieve the ROS in SCs and alleviate oxidative stress.It was concluded that albiflorin can increase the proliferation of SCs and improve the state of oxidative stress with the protection of SCs.
ABSTRACT
Objective To establish the HPLC fingerprints of Tangbikang Granules; To scientifically evaluate and effectively control the quality of Tangbikang Granules; To ensure its production stability. Methods HPLC was performed on the column of Germany Merck RP-18 endcapped (250 mm × 4.6 mm, 5 μm) with mobile phase of acetonitrile-0.1% formic acid water; column temperature was 40 ℃; flow rate was 1.0 mL/min; detection wavelength was 240 nm; volume injection was 20 μL. Fingerprint Similarity Evaluation Software (edition 2004A) of Chinese Pharmacopoeia Commission was used to evaluate the similarity of the 10 batches of Tangbikang Granules, and to analyze the correlations of 9 ingredients in Tangbikang Granules. Results Wogonoside was used as the reference peak, and the common mode for the HPLC fingerprints was set up. The similarities of the 10 batches of Tangbikang Granules were above 0.930, and altogether 25 common peaks in the chromatograms were found, of which 18 peaks were assigned to Chinese materia medica in Tangbikang Granules. Conclusion The method has good separability and is accurate and simple, which can provide references for the quality control of Tangbikang Granules.