2.Inhibiting effects of three components of Astragalus membranaceus on oxidative stress in Chang Liver cells.
Jian LI ; Lin HAN ; Yu-fang MA ; Yi-fan HUANG
China Journal of Chinese Materia Medica 2015;40(2):318-323
The main objective of this research is to investigate the effects of astragaloside IV, calycosin separately glucoside, formononetin on oxidative stress in Chang Liver cells induced by H2O2. In the experiments, Chang Liver cells (a kind of normal human hepatocytes) were used as the research object, bifendate which has a clear hepatoprotective effect was used as the positive control drug, then the oxidative damage model of Chang Liver cells were established by H2O2. Cells were divided into six groups: blank control group, oxidative stress group, astragaloside IV group, calycosin separately glucoside group, formononetin group and positive control group. Then endogenous antioxidant system related indexes were detected by micro plate and colorimetric method; intracellular reactive oxygen species (ROS) were detected by DCFH-DA fluorescent probe; and the expressions of CYP2E1 were evaluated by liver microsomes, mRNA, and protein, respectively with spectrophotometry, Real-time PCR method, and Western blot technique. Results showed that H2O2 decreased antioxidant activity, and increased ROS level and expression of CYP2E1. The above oxidative stress status had been changed with protections of the three components of Astragalus membranaceus (compared with oxidative stress group, P < 0.05, P < 0.01), which taken as a whole had equivalent effects as the drug of positive control group( bifendate). Taken together, three Astragalus membranaceus ingredients all had significant or extremely significant inhibiting effects on oxidative damaged Chang Liver cells which were induced by H2O2, and the oxidative damage of Chang Liver cells had been relieved.
Astragalus membranaceus
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chemistry
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Cells, Cultured
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Cytochrome P-450 CYP2E1
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metabolism
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Humans
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Isoflavones
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pharmacology
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Liver
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drug effects
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Oxidative Stress
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drug effects
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Reactive Oxygen Species
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metabolism
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Saponins
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pharmacology
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Triterpenes
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pharmacology
3.Content Determination of Chlorogenic Acid,Protocatechuic Acid and Total Phenolics in Chaenomeles sinersis from Different Areas
Yu HAN ; Mo YANG ; Xiaomei XIE ; Jingjing CHENG ; Jian WANG
China Pharmacy 2015;(24):3399-3402,3403
OBJECTIVE:To establish a method for the content determination of chlorogenic acid,protocatechuic acid and total phenolics in Chaenomeles sinensis,compare the content of total phenolics and 2 phenolic acids from different areas. METHODS:Wavelength switching HPLC method was conducted to determine the chlorogenic acid and protocatechuic acid. The column was Shim-pack CLC-ODS(M)with the mobile phase of acetonitrile-0.1% phoephoric acid(15∶85,V/V),the detection wavelength was 259 nm(for protocatechuic acid)and 325 nm(for chlorogenic acid)and the switching time was 14 min. With the index of proto-catechuic acid,Folin-Ciocalteau colorimetric method was conducted to determine the total phenolics. RESULTS:The 2 methods of quantitative analysis showed that the precision,repeatability,recoveries and standard curves were all validated by methodology. The mass fraction of total phenolics was 0.87%-3.77% with the average of 2.16%;the chlorogenic acid was 0.053%-0.387% with the average of 0.192% and the protocatechuic acid was 0.024%-0.541% with the average of 0.087%. The order of total phenolics content in C. sinensis from different areas was Yunnan>Anhui Xuancheng>Sichuan>Hubei and the order of total amount of chlo-rogenic acid and protocatechuic acid from different areas was the same as the total phenolics. There were differences among the con-tents of C. sinensis from different areas,however,the positive correlation was found between the content of total phenolics and the total amount of chlorogenic acid and protocatechuic acid with the pearson correlation coefficient of 0.719(P<0.01).CONCLU-SIONS:The established method is simple,accurate and reproducible and can be used for the content determination of chlorogenic acid,protocatechuic acid and total phenolics in chaenomelis fructus.
4.The mechanism of hyperintense signal at interpeduncle region on diffusion weighted imaging
Hongbin HAN ; Jian WANG ; Junhao YAN ; Jincai PI ; Yu DOU
Chinese Journal of Radiology 2008;42(9):936-940
Objective To study the mechanism of hyperintensed signal at interpeduncle region in meseneephalon on DWI with diffusion sensitive gradient(Gd)applied perpendicular to the slice direetion (DWIs).Methods MRI scanning was centered at interpeduncle region.and the protocols were as follows:(1)With electrocardiograph(ECG)triggering,DWIs was acquired during systole or diastole periods respectively.(2)To observe the shape of hyperintense signal,DWIs with various slice directions was applied,which was set parallel to skull base,along Y axis,or along the oblique line between Y and Z axis with 30°to Z axis.(3)Fiber tracking was performed with the hyperintense signal as"seed"region,fractional anisotropy(FA)and ADC were derived from(multiple directions diffusion weighted imaging,MDDW).FA and ADC of hyperintease region were compared with those of the nearby fiber by using pair t-tesL Results Signal intensitv of hyperintense region on DWIs were 2296.28±38.19 and 153.81±37.91 for b=0 and b=1000 at systole period.while they were 295.36±38.84 and 154.03±37.52 at diastole period.No staffstical difference wag demonstrated between them(t=1.34,0.62,P>0.05).The ADC valus on DWIs acquired during systole and diastole periods were(6.07±2.20)×10-4and(6.69±1.44)×10-4 mm2/s respectively.There was rio statistical difference between them(t=0.94,P>0.05).DTI fiber tracking verified that the hyperintense region located at the decussation of superior eerebelum peduncle(SCP)in mesencephalon.It was in long and narrow heart shape or rectangular shape on DWIs.The shape depended on the direction of Gd The ADC value derived from MDDW at hyperintense region and the fiber below were(10.61±3.42)×10-4 and(9.24±2.21)×10-4 mm2/s respectively.No statistical difference was demonstrated between them(t=0.61.P>0.05).While,the FA at hyperintensed region 0.43±0.13 was less than that of the fiber below 0.61±0.08(t=8.32.P<0.05).Conclusions The shape of the hyperintense region on DWIs depends on the direction of Gd.The cardiac period has no effects on the hyperintense signal at peduncles,which may be attributed to the anisotropy of SCP decussation in mesencephalon.
7.Diagnostic value of radionuclide imaging combined with routine CT in detecting hepatic focal nodular hyperplasia
Xue-min, L(U) ; Shu-hong, YU ; Jian-kui, HAN
Chinese Journal of Nuclear Medicine 2011;31(4):250-254
Objective To investigate radionuclide imaging and routine CT in diagnosing hepatic focal nodular hyperplasia (FNH) and the combined diagnostic value of the two modalities. Methods Thirty-two patients with hepatic FNH were retrospectively studied. All patients underwent routine CT scan. Twenty-four patients were examined by 99Tcm-sulfur colloid (SC) hepatic planar scintigraphy and SPECT/CT imaging, and then patients who had abnormal foci underwent 99Tcm-diethyl iminodiacetic acid (EHIDA) triple-phase hepatobiliary imaging. x2 -test of four-table or Fisher exact probabilities in 2 × 2 table was applied for statistical analysis. Results Of all 32 patients pathologically diagnosed as FNH with single solitary nodule, 25 were classified as classic type and the rest 7 as non-classic type. Although routine CT found all hepatic lesions, only 15 cases were diagnosed pathologically as FNH classic type but the rest were either misdiagnosed or left as indeterminate. On radionuclide imaging (hepatic colloid scintigraphy plus triple-phase hepatobiliary images), 11 patients with big foci (with maximal diameter >3 cm) out of 24 patients were correctly diagnosed as FNH, with 7 diagnosed as classic type FNH and 4 as non-classic. Other 13 patients were either misdiagnosed or simply missed. The diagnosing rates of routine CT and radionuclide imaging were60.0% (15/25) and 38.9% (7/18) for FNH classic type, 0/7 and 4/6 for non-classic type,50.0% (10/20) and 73.3% (11/15) for big foci, 41.7% (5/12) and 0/9 forsmall foci (with maximal diameter≤3 cm), respectively. The total diagnosing rate of radionuclide imaging combined with routine CT was significantly higher than that of routine CT or radionuclide imaging alone ( x2 = 4. 48, P < 0. 05;x2 =4.27, P <0.05 ). Conclusion Radionuclide imaging in combination with routine CT may improve the diagnostic accuracy for hepatic FNH patients.
8.Effect of Electroacupuncture on the Expression of Adenosine A1Receptor in Hypothalamus and Spinal Cord of Adjuvant Arthritis Rats
Jing HAN ; Jian WANG ; Xiaohua YU ; Fudong WU
Shanghai Journal of Acupuncture and Moxibustion 2016;(1):71-75
Objective To observe the effect of adenosine A1 receptor on pain and electroacupuncture analgesia, and to explore the action mechanism of electroacupuncture in analgesia.Method Adjuvant arthritis rats were taken as the study subjects. 24 rats were randomized into a normal group, a model group and an electroacupuncture (EA) group, 8 rats in each group. The pain threshold was evaluated by using thermal radiation method, immunohistochemical method and real-time fluorescence quantitative PCR were adopted to observe the expression of adenosine A1 receptor in hypothalamus and spinal cord.Result One day after modeling, the pain thresholds of right hind paw in the model group and EA group were significantly changed compared to that before modeling in the same group (P<0.01). The pain thresholds of right hind paw in the model group and EA group were significantly different from that in the normal group one day after modeling (P<0.01). 7 d after modeling, the pain threshold of right hind paw in the model group was still significantly lower than that before modeling in the same group (P<0.01), and it was significantly different from that in the normal group and EA group (P<0.01). The pain threshold was significantly enhanced in the EA group 7 d after modeling, and was significantly different from that of 1 d after modeling in the same group (P<0.01). The positive cell expression was lower in the model group and was significantly different from that in the normal group (P<0.01). The expression of adenosine A1 receptor in the EA group was markedly higher than that in the model group (P<0.01). The expression of adenosine A1 receptor in hypothalamus and spinal cord of the model group was significantly lower than that of the normal group (P<0.01). The expression of adenosine A1 receptor in hypothalamus and spinal cord of EA group was markedly higher than that of the model group (P<0.01).Conclusion EA can up-regulate the expression of adenosine A1 receptor in hypothalamus and spinal cord of adjuvant arthritis rats.
9.Hyperbaric oxygen for suppressing glial scar formation and inflammation after a stab wound to the cerebral cortex
Na YIN ; Yu WANG ; Yuanyuan HAN ; Huiyan HE ; Jian ZUO
Chinese Journal of Physical Medicine and Rehabilitation 2013;(6):442-446
Objective To observe any influence of hyperbaric oxygen (HBO) treatment on the formation of glial scars,and to explore how HBO suppresses the inflammatory reaction to injury.Methods A total of 96 healthy,adult,male,Sprague-Dawley rats were used to model cerebral puncture injury.They were then randomized into a control group and a treatment group,with 48 rats in each group.The treatment group received HBO treatment,while the control group received no special treatment.At 1,3,7,14 and 28 days after the puncture injury,the rats' right brain tissues were harvested and immunohistochemical staining was employed to compare the changes in number of astrocytes and microglial cells around the injury in the two groups.The level of tumor necrosis factor α (TNF-α) and interleukin 1 β (IL-1β) in the cerebral tissue was examined using ELISA.Results Among the control group the average wound areas after 7,14 and 28 days were (2.73 ± 0.05)μm2,(3.42 ± 0.18)μm2 and (2.41 ± 0.09) μm2,a significant reduction after 28 days compared with 7 and 14 days.The corresponding average wound areas of rats in the treatment group were (2.78±0.12)μm2,(2.59 ±0.08)μm2 and (1.20 ±0.06)μm2.There the average wound area had decreased significantly after 14 days,and the further reduction after 28 days was also significant.The numbers of GFAP-positive astrocytes at 14 and 28 days had increased significantly compared with after 7 days in both the control group and the treatment group.The average number of GFAP-positive astrocytes in the control group at 28 days had decreased significantly compared with after 14 days.Compared with the control group at the same time points,the number of GFAP-positivc astrocytes in the treatment group was significantly less.After modeling,the number of ionized calcium-binding adapter molecule Ⅰ (Ibal)-positive microglial cells increased significantly,but there was a significant decrease in both the control and treatment groups by 7 days.The average number of Ibal-positive microglial cells in the treatment group was significantly less than in the control group at all of the time points.Compared with the first day after modeling,the TNF-α concentration of the controls at 3 and 7 days was significantly higher,but by the 7th day it was significantly lower than it had been after 3 days.The average IL-1β concentration in the control group and TNF-α concentration in the treatment group had increased by day 3,but then decreased by day 7.The IL-1β concentration of the treatment group declined gradually.The average TNF-α and IL-1 β concentrations of the treatment group were significantly lower than those of the control group at all of the time points.Conclusion HBO treatment has a relatively good curative effect on cerebral puncture injury.It can accelerate wound healing and reduce the formation of glial scars.Its mechanism could be related to the deactivation of astrocytes and microglia cells and reducing the levels of cell factors that promote inflammation.