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OBJECTIVE To investigate the effects and mechanism of polysaccharides from Hedyotis diffusa （HDP） on isoniazid （INH）-induced liver injury. METHODS Healthy transgenic zebrafish with liver-specific fluorescence were divided into normal group， model group （4 mmol/L INH）， HDP low-concentration group （4 mmol/L INH+50 mg/mL HDP） and HDP high- concentration group （4 mmol/L INH+100 mg/mL HDP）. After grouping treating， the liver fluorescence area， fluorescence intensity and pathological changes of liver tissue were observed. Human liver L02 cells were divided into normal group， model group （4 mmol/L INH）， HDP low-concentration group （4 mmol/L INH+2 mg/mL HDP）， and HDP high-concentration group （4 mmol/L INH + 4 mg/mL HDP）. After grouping treating， the cell viability was detected， and the levels of alanine transaminase （ALT）， aspartate transaminase （AST）， and the content of glutathione （GSH） as well as the expression levels of silent information regulator 1 （Sirt1）， nuclear factor-erythroid 2-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1） and quinone oxidoreductase 1 （NQO1） proteins were detected. RESULTS Compared with the model group， the HDP low- and high-concentration groups showed varying degrees of increase in the fluorescence area and fluorescence intensity （except for HDP low-concentration group） of zebrafish liver （P＜0.05 or P＜0.01）， and the characteristics of liver injury and necrosis had been improved to varying degrees. Compared with model group， the survival rate of L02 cells， the content of GSH （except for HDP low-concentration group）， the protein expression levels of Sirt1 （except for HDP low-concentration group）， Nrf2， NQO1， HO-1 （except for HDP low-concentration group） were significantly increased in HDP low- and high-concentration groups （P＜0.05 or P＜0.01）， and the levels of ALT and AST （except for HDP low-concentration group） were significantly decreased （P＜0.05）； the number of survival cells significantly increased， while the number of damaged or dead cells significantly decreased. CONCLUSIONS HDP has a potential protective effect against INH-induced liver injury， the mechanism of which may be associated with activating Sirt1/Nrf2 signaling pathway， improving mitochondrial function and enhancing antioxidant capacity.
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OBJECTIVE：To establish HPLC fingerprint of different products of suet oil-baked Epimedium brevicornum ，and to screen the optimal baking technology. METHODS ：HPLC method was adopted. Using icariin as reference ，HPLC fingerprints of 22 batches of samples were drawn. The similarity was evaluated by using Similarity Evaluation System of TCM Chromatographic Fingerprint（2012 edition），and common peaks were confirmed. HCA ，PCA and OPLS-DA analysis were performed by SIMCA 14.1 statistical software. Taking variable importance in the project ＞1 as criteria ，biomarkers affecting the quality difference of suet oil-baked E. brevicornum were screened ；using mass marker as index ，the baking technology was screened by baking technology. RESULTS：There were 18 common peaks in 22 batches of samples. The similarities were between 0.831 and 0.991. Totally 7 common peaks were identified ，i.e. epimedin A ，epimedin B ，epimedin C ，icariin，sagittatoside A ，sagittatoside B ，baohuoside Ⅰ. The 22 batches of samples were clustered into two categories ，S19-S22 were clustered into category Ⅰ and S 1-S18 were clustered into category Ⅱ. The category Ⅱ was sub-clustered into category Ⅱa（S15-S18），category Ⅱb（S10-S14），category Ⅱc （S1-S9）；the result of PCA analysis was consistent with above results. OPLS-DA showed that the biomarkers affecting the quality difference were icariin ，sagittatoside B and baohuoside Ⅰ. The results of kinetic studies showed that the content of icariin when baked at 180 ℃ for 25 min or 190 ℃ for 20 min，that of baohuoside Ⅰ when baked at 180 ℃ for 30 min or 190 ℃ for 15 min and that of epimedin B when baked at 210 ℃ for 18 min were the highest ；according to above results ，the optimal baking technology was baking at 180 ℃ for 25-30 min or 190 ℃ for 15-20 min. CONCLUSIONS ：Established fingerprint is stable ， reliable and reproducible. The multivariate statistical analysis can be used for the changes of chemical components in E. brevicornum under different baking condition and preliminary selection of baking technology.
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Objective:To discuss the importance and significance of training base establishment for clinical pharmacy of traditional Chinese medicine ( TCM) . Methods:The urgency and necessity of training base establishment for TCM clinical pharmacy was stated based on the status and problems of TCM clinical pharmacy, and the important significance of training base establishment of TCM clini-cal pharmacy was clearly put forward. Results:The establishment of TCM clinical pharmacy training base with the aim of training pro-fessional talents for TCM clinical pharmacy can provide clinical pharmaceutical care for patients with higher quality, which surely will promote the development of TCM clinical pharmacy and the pharmaceutical care level. Conclusion:It is extremely urgent and signifi-cant to set up TCM clinical pharmacy training base, which is the important topic needed to be solved in TCM clinical pharmacy work.
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Objective To study and compare the metabolism of Hedyotis diffusa flavonoid extract in intestinal flora from human, normal rats and pseudo germ-free rats in vitro. Methods Intestinal flora of human, normal rats and pseudo germ-free rats were incubated with Hedyotis diffusa extract, rutosid, isoquercitrin and quercitrin under anaerobic conditions at 37℃.High-performance liquid chromatography with diode-array detection ( HPLC-DAD ) was used for the analysis of components and metabolic products.The trends of metabolic transformation of the various components were analyzed according to the changes in chromatographic peak areas at different incubation time points. Results The components of Hedyotis diffusa extract and its monomer were quickly metabolized by human and normal rat intestinal flora and the main metabolite was quercetin.However, they were hardly metabolized by intestinal flora from pseudo germ-free rats. The metabolism effect of intestinal flora was weaker on components than on the corresponding monomers. Conclusion By comparing flavonoids metabolism in intestinal flora from human, normal rats and pseudo germ-free rats, the important role of intestinal microflora in the metabolism of flavonoids is further confirmed.
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Objective To explore an UPLC method for simultaneous content determination of the five nucleosides (cytidine, uridine, adenine, thymidine and adenosine) in Hedyotis diffusa and its adulterants; To compare the content differences.Methods The analysis was performed on a BEH C18 column (2.1 mm×50 mm, 1.7 μm) by UPLC eluted with acetonitrile and water in gradient mode. The flow rate was 0.5 mL/min; the detection wavelength was set at 254 nm; the column temperature was set at 30℃.Results Five nucleosides have good linear relationship, precision, stability, and repeatability according to the requirements of the methodology determination. The recoveries were among 98.7%–101.5%. Five nucleosides in Hedyotis diffusa and its adulterants from different areas were determined by the UPLC method.Conclusion The method is certified to be simple, rapid, accurate and reliable, which can be used for the determination of nucleosides in Hedyotis diffusa and its adulterants.
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Herba Hedyotis Diffusae has many kinds of pharmacological activities such as anti-tumor, antioxidant, immune regulation and so on. This article collected and reported the related literature at home and abroad in recent years, found that pharmacological active ingredients of Herba Hedyotis Diffusae show consistency of polysaccharides and flavonoids to varying degrees, and summarized the pharmacological effects and extraction and purification process. The author believed that, the study about Herba Hedyotis Diffusae has made relatively great achievements, but the research mainly focused on polysaccharides or flavonoids, and efficacy of Herba Hedyotis Diffusae was not in enough in-depth exploration. The indicator of extraction and purification process is relatively single. This study estimated the mechanism of Herba Hedyotis Diffusae from the aspects of absorption and metabolism of intestinal flora and relationship between medicine structure and exert efficacy, hoping to optimize the extracting processes by multiple indexes, and provided references for further exploitation and utilization of Herba Hedyotis Diffusae.
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Objective To build a method for content determination of Hypericin in Guanye Jinsitao drop pills by HPLC.Methods The column was Kromasil C18 (150 mm×4.6 mm, 5μm), mobile phase was acetonitrile-0.1% phosphoric acid (16∶84), UV detection was performed at 360 nm, the flow rate was 1.0 mL/min, and the detector drift tube temperature was set at 30℃.Results The calibration curves were linear in the range of 0.132-1.188μg for Hypericin (r=0.999 9). The average recovery was 99.55% (RSD=0.78%).Conclusion The method is simple, accurate, reliable, and can be used for the quality control of Hypericin in Guanye Jinsitao drop pills.
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Objective To develop an assay of HPLC for the content determination of dryocrassine in Guanye Jinsitao drop pills. Methods The column was Kromasil-C18 (150 mm×4.6 mm, 5μm);mobile phase was acetonitrile-chloroform-isopropanol-0.1% phosphoric acid (52∶8∶30∶10);flow rate was 1.0 mL/min;UV detection was performed at 285 nm;detected drift tube temperature was set at 25 ℃. Results The calibration curves were linear in the range of 0.236-2.124 μg for dryocrassine (r=0.999 9). The average recovery rate of dryocrassine was 99.64% (RSD=1.90%). Conclusion The method is simple and accurate. It is suitable for the determination of dryocrassine in Guanye Jinsitao drop pills.