1.Simultaneous determination of five nucleosides in Houtoujun Tablets by RP-HPLC
Haiming TANG ; Deqin WANG ; Huiye ZHANG
Chinese Traditional Patent Medicine 2017;39(5):964-967
AIM To establish an RP-HPLC method for the simultaneous content determination of five nucleosides in Houtoujun Tablets (mycelia of Hericium erinaceum).METHODS The analysis of aqueous extract of this drug was performed on a 26 ℃ thermostatic Zorbax SB-Aq C18 column (250 mm ×4.6 mm,5 μm),with the mobile phase comprising of methanol-0.1% phosphoric acid flowing at 0.5 mL/min in a gradient elution manner,and the detection wavelength was set at 260 nm.RESULTS Cytidine,adenine,uridine,adenosine and guanosine showed good linear relationships within the ranges of 0.007-0.169,0.005-0.115,0.032-0.797,0.032-0.792 and 0.004-0.103 μg,whose average recoveries (n =6) were 99.4% (RSD =2.0%),98.3% (RSD =1.8%),98.0% (RSD =2.8%),101.4% (RSD =1.9%) and 101.7% (RSD =1.2%),respectively.CONCLUSION The contents of five nucleosides in Houtoujun Tablets from different manufacturers exhibit obvious differences,to which we should pay attention.
2.Cardioprotection of orientin against myocardial ischemia reperfusion injury through induction of autophagy
Liya LIU ; Huiye WANG ; Xiulan HUANG
Chinese Pharmacological Bulletin 2016;32(4):542-547
Aim To investigate the role of autophagy in the cardioprotection by orientin and the relative molec-ular mechanisms in cell apoptosis and autophay. Meth-ods The isolated ischemia reperfusion ( I/R ) heart model was built firstly. The Experiments were divided into seven groups:control group, I/R group, different concentrations of orientin-treated group ( 1. 0 mg · kg-1 ,2. 0 mg· kg-1 ,4. 0 mg · kg-1 ) , autophagy in-hibitor group and resveratrol group. Hemodynamic in-dex were recorded by PowerLab, the activities of myo-cardial enzymes were detected through Biochemical an-alyzer, the ultrastructure changes and autophagosomes in myocardial cells were detected by electron microsco-py, the apoptosis was detected by TUNEL, and LC3 and Beclin1 protein levels of left ventricle were meas-ured by Western blot. Results Orientin at middle and high concentrations(2 and 4 mg·kg-1 ) induced auto-phagy shown by increased the number of autophago-somes, LC3-Ⅱ/LC3-Ⅰ ratio and Beclin 1 expression after ischemia-reperfusion. The induction of autophagy by orientin was correlated with enhanced cardiac func-tion and decreased apoptosis. But wortmannin, a kind of autophagy inhibitor, significantly attenuated the ori-entin-induced autophagy and increased apoptosis. Con-clusion The cardioprotection of orientin against myo-cardial ischemia reperfusion injury may be mediated through the inhibition of apoptosis and induction of au-tophagy.
3.Differentiation on Original Plants of Xi-Huang-Cao through DNA Barcodes
Huiye ZHANG ; Feng LIU ; Deqin WANG ; Ruoting ZHAN ; Xinye MA ; Weiwen CHEN
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(7):1487-1490
This study was aimed to identify original plants of Xi-Huang-Cao (XHC) through DNA barcodes. The nu-cleotide sequence information of rbcL, psbA-trnH, matK and ITS2 regions were abstracted using standardized man-ners from 41 samples (including Rabdosia serra, R. lophanthoides and R. lophanthoides var. graciliflora). Sequencing efficiency of each marker was calculated. Species identification capability was tested on the basis of TaxonGap. The results showed that sequence success rates were 100% for rbcL, 90.2% for psbA-trnH, 87.8% for ITS2, and 70.7%for matK. Three markers (rbcL, psbA-trnH and ITS2) were competent for species discrimination (not for subspecies). It was concluded that rbcL can be the preferred barcode for XHC because of its convenience and efficacy.
4.Establishment and comparison of 2 mouse models of kainic acid induced epilepsy
Can TENG ; Meiling YU ; Guohui JIANG ; Huiye FAN ; Faxiang WANG
Journal of Army Medical University 2024;46(7):661-669
Objective To investigate and analyze the behavioral and pathological differences in early-stage mouse models of epilepsy established by 2 different administration routes for kainic acid(KA),intracerebroventricular(ICV)injection and intraperitoneal(IP)injection.Methods A total of 100 male C57BL/6N wild-type(WT)mice(20~22 g)were randomly divided into ICV+normal saline(NS)control group(n=10),ICV+KA model group(n=40),IP+NS control group(n=10)and IP+KA model group(n=40).The ICV+KA model group was given 600 nL of KA(0.5 mg/mL)via ICV injection,and the IP+KA model group was injected with different dose of KA(25 mg/kg).Two control groups were administered equal volumes of NS via corresponding routes.After 3 d of modeling,the evaluation of behavioristics,molecular biology(including Western blotting),and neuropathological assessments(including FJB staining,TUNEL staining and immunofluorescence staining)were performed.Results No epileptic seizures were observed in both 2 control groups,while exhibited seizures were observed in both model groups.The mortality rates of the IP+KA group and the ICV+KA group were 47.50%and 65.00%respectively,while the success rates of modeling were 80.00%and 60.00%respectively.Compared with the IP+KA group,the ICV+KA group showed a significant increase in success rate and a significant reduction in mortality rate.FJB and TUNEL staining results showed that,compared with the IP+KA group,the severity of neurodegeneration and apoptotic changes in the hippocampus of the ICV+KA group were more significant(P<0.05).Compared with the IP+KA group,there was also a significant difference in the expression of apoptotic proteins in the hippocampus of the ICV+KA group(P<0.05).Immunofluorescence results showed that the astrocytes and microglia in the hippocampus and cortex of the ICV+KA and IP+KA groups were significantly activated compared with the control groups(P<0.05),but the activation of glial cells in the hippocampus and cortex of the ICV+KA group was stronger than that of the IP+KA model group(P<0.05)and the activation levels in the ICV+KA group were higher than in the IP+KA model group(P<0.01).Moreover,expression levels of GFAP and Iba-1 proteins in the hippocampus and cortex were higher in the ICV+KA group than the IP+KA group(P<0.05).Conclusion Two routes of KA administration are effective in construct epilepsy models.The mice with ICV administration route show a higher success rate and lower mortality rate,and more significant neuropathological damage and glial cell activation.