1.Effect of genistein on lipid accumulation in H4Ⅱ E cell induced by oleic acid
Lingling QIN ; Tunhai XU ; Jingxin ZHOU ; Tonghua LIU
International Journal of Traditional Chinese Medicine 2013;(7):600-603
Objective To observe the effects of genistein(GEN)on oleic acid(OA)induced lipid accumulationin in H4 ⅡE cells and to discuss the possible mechanism of GEN in the pointof AMPK.Methods H4ⅡE cells were cultured in vitro.The control group(NOR),OA treatment group(MOD group)and GEN treatment group were established according to the experimental requirements.The effects of GEN on the proliferation of H4 Ⅱ E cells were measured with MTT assay.The intraeellular TG mass was quantified spectrophotometrically using TG test kit.Cell protein was determined by DCTM Protein Assay kit.The intracellular TG concentration which was used to evaluate lipid accumulation was corrected using protein content as an internal standard.Western blotting was applied to determine the expression of AMPK and P-AMPK (Thr172).Accordingly the phosphorylation levels of AMPK was by means of P-AMPK(Thr172)/AMPK.Results OA treatment can induce lipid accumulation in H4 Ⅱ E cells while GEN treatment can decrease the intracellular TG concentration through up-regulate the phosphorylation levels of AMPK,though the effect was blocked by Compound C that is the inhibitor of AMPK.Conclusion GEN has anti-accumulation of lipid effect in H4ⅡE cell.The mechanism of GEN protective effect is partially due to AMPK.
2.Effect of Gymnema Sylvestre on Protein Kinase B in Adipose Tissues of Insulin Resistance Mice
Lingling QIN ; Tonghua LIU ; Xiaohong MU ; Tunhai XU ; Wen SUN
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(5):1042-1047
This article was aimed to study effects and mechanisms of Gymnema sylvestre on protein kinase B (PKB) and its phosphorylation in adipose tissues of KKAy mice which were mainly characterized by insulin resistance (IR). A total of 18 KKAy mice were randomly divided into the diabetes model (DM) group and Gymnema sylvestre (GS) group according to body weight levels. And 9 normal C57BL/6J mice were used as the normal control (NC) group. Intragastric administration of medication was given to mice for 8 weeks. At the end of the experiment, all animals were tested for fasting plasma glucose (FPG) and fasting insulin level (Fins) for evaluation of insulin sensitivity index (ISI). Expressions of phosphoinositide-dependent kinase-1 (PDK1), PKB, P-PKB (Ser473), P-PKB (Thr 308) in adi-pose tissues of epididymis were determined. The expression of phosphatase and tensin homolog (PTEN) mRNA was also determined. The results showed that compared with the DM group, the GS group showed lower FPG and Fins, higher ISI. The expression of P-PKB (Ser473) phosphorylation and P-PKB (Thr 308) were increased, and the PDK1 and PTEN mRNA were decreased. It was concluded that GS can improve insulin sensitivity of KKAy mice through activating PKB by up-regulate the expression of P- PKB (Ser473) and its phosphorylation ratio and P- PKB (Thr 308) in adipose tissues.
3.Research on Etherification Reaction of Three Kinds of Isoflavones from Chickpeas
Bo LI ; Yongqiao LIU ; Haoxia SHI ; Tunhai XU ; Tonghua LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(5):786-790
This study was aimed to improve the drug activity of three kinds of isoflavones from Chickpeas.Biochanin A,formononetin and genistein were used as raw materials.Acetone was used as solvent.Potassium carbonate was used as catalyst.The etherification reaction was with 1,3-dibromopropane,1-bromopropane and 3-bromopropene.The results showed that 9 isoflavone ramifications were synthesized.This method was simple,easy to control with high yield.It was concluded that the product structure was confirmed by 1H-NMR,13C-NMR and ESI-MS analysis.It laid a foundation for the structural study basis in the further research of its drug activity.
4.Study on Synergistic Hypoglycemic ActivityIn Vitroof Three Kinds of Isoflavone in Chickpea and Its Derivatives of Ethoxylation
Pengshou LI ; Ying WEI ; Xiaojuan SHI ; Bo LI ; Dongchao WANG ; Tunhai XU ; Tonghua LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(6):1176-1183
Through the ethoxylation structure modification of three isoflavones which were genistein, biochanin A and formononetin in chickpea, the hypoglycemic activity and the synergistic hypoglycemic activity were studied. Ethoxylation structure modification was given on three kinds of isoflavones. The hypoglycemic activity of three kinds of isoflavones and their derivatives were studied. The synergistic hypoglycemic activity of the compounds was also studied. The insulin-resistance HepG2 cell was selected as the model of hypoglycemic activity screening. The results showed that four ethoxylation products were synthesized. And the hypoglycemic activity of genistein was better than biochanin A and formononetin with significant difference (P < 0.05). The maximum dosage of three isoflavones was compared with the positive drug metformin hydrochloride. And the effect was not as good as metformin hydrochloride. There was significant difference (P < 0.05, orP < 0.01). There was no significant difference of effect between the compound a and the compound d (P > 0.05). Effects of the compound b and compound c were not as good as the compound a and compound d. There was statistical difference (P < 0.05). The activity of compound d was significantly improved than the parent compound (P < 0.05). There was no statistical difference on the hypoglycemic activity between combination 5 and metformin hydrochloride (P > 0.05). It was concluded that the combinations of compounds played synergistic effects with better hypoglycemic effect. It provided a basis for developing hypoglycemic agents with the intellectual property rights.
5.Research on Content Determination Methods ofTang-Nai-Kang Granules
Ying WEI ; Xiaojuan SHI ; Dongchao WANG ; Pengshou LI ; Bo LI ; Linyi LI ; Tunhai XU ; Tonghua LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(6):1169-1175
This article was aimed to study the content determination methods ofTang-Nai-Kang (TNK) granules. The content of rosemary acid in TNK granules was determined by HPLC method, using C18 chromatographic column (150 mm×4.6 mm, 5μm), the mobile phase of 0.1% trifluoroacetic acid - methanol (60:40),λ of 330 nm. The theoretical plate number was more than 2 000. The standard curve method was used in the determination. Contents of ginsenosides Rg1, Re and Rb1 were determined by HPLC method, using C18 chromatographic column (150 mm× 4.6 mm, 5μm), the mobile phase of water-acetonitrile, gradient elution method,λ of 203 nm. The theoretical plate number was more than 2 000. The standard curve method was used in the determination. The results showed that rosmarinic acid was linear in the range of 0.300-0.500 mg·mL-1 with good linearity. The regression equation wasY = 1E + 07X -7 334,R2 = 0.999. Its sample recovery was 97.32% (RSD 1.25%). Ginsenoside Rg1 was linear in the range of 0.138-0.220 mg·mL-1 with good linearity. The regression equation wasY = 2E + 06X +34 864,R2 = 0. 999. Ginsenoside Re was linear in the range of 0.126-0.250 mg·mL-1 with good linearity. The regression equation wasY = 2E + 06X + 39 879,R2 = 0. 999. Ginsenoside Rb1 was linear in the range of 0.132-0.220 mg·mL-1 with good linearity. The regression equation wasY = 2E + 06X + 39 070,R2 =0. 999. Their sample recoveries were Rg1 97.97% (RSD 1.83%), Re 101.80% (RSD 1.83%), Rb1 98.35% (RSD 1.82%). It was concluded that the content determination method established for the quality control of TNK granules was simple, fast and reliable.
6.Study on Extraction Conditions of Total Flavonoids from Fruit ofVernonia esculenta Hemsl.
Meili SUN ; Lei LI ; Heyu WANG ; Jing ZHANG ; Tunhai XU ; Tonghua LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(5):1108-1112
Spectrophotometry and single factor test were used in the investigation of effects on extraction results of total flavonoids from the fruit ofVernonia esculenta Hemsl. from aspects of ethanol concentration, frequency of extraction, length of extraction time, and the solid-liquid ratio. The extraction conditions were optimized by orthogonal test. The results showed that the optimum extraction conditions were ethanol concentration of 70%, extracted for 4 times, 1 h for each time, and the solid-liquid ratio of 1:20. Under these extraction conditions, the content of total flavonoids in the fruit ofVernonia esculenta Hemsl. was 103.55 mg·g-1.
7.Determination of Three Xanthone Glycosides in Gentianella Acuta by HPLC
Qian WEI ; Tonghua LIU ; Xiaojuan SHI ; Pengshou LI ; Ying WEI ; Tunhai XU
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(5):1060-1064
To develop a method for determining 3 Xanthone glycosides (1, 5-dihydroxy- 3-methoxyxanthone 8-O-β-D-glucopyranoside(F1), 1-hydroxy-3, 4-dimethoxyxanthon 7-O-β-D-glucopyranoside (F2), 1, 8-dihydroxy-3, 4-dimethoxyxanthone 5-O-β- D-glucopyranoside (F3) of Gentianella acuta by HPLC. The Thermo syncronis C18 (4.6 mmí250 mm, 5 μm) was used for simultaneous determination of 3 Xanthone glycosides in G. acuta. Isocratic elution with water and acetonitrile was 75.5:24.5. The flow rate was 1.0 mL·min-1 and the detection wavelength was set at 254 nm. The column temperature was 35℃. The linear concentration ranges of F1,F2,F3 were 14.06 ~ 281.28 μg·mL-1 (R2=0.999 7),0.56~11.16 μg·mL-1 (R2=0.999 8),0.46~9.20 μg·mL-1 (R2=0.999 9), respectively; The average recov-eries (n = 9) were 99.70% (RSD=1.06%),99.78% (RSD=1.21%),100.28% (RSD=1.15%), respectively. The method is simple, accurate and sensitive, and can be used for the quality control of G. acuta as a reference.
8.Simultaneous Determination of Three Flavonol Glycosides in the Stems and Leaves of Tribulus terrestris L . by HPLC
Haishui GUI ; Tonghua LIU ; Lingling QIN ; Pengshou LI ; Chunna LI ; Yangyang LIU ; Tunhai XU
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(5):1055-1059
In this paper, three higher content of flavonol glycosides from the stems and leaves of Tribulus terrestris L. were determined by reversed phase HPLC. The results showed that there was good liner relationship between the peak areas and the sample concentration at the ranges of 0.011 2~0.280 0 μg (r=0.999 6), 0.064 8~2.592 0 μg (r=0.999 8) and 0.018 4~0.460 0 μg (r=0.999 8) for isorhamnetin-3-O-β-D-gentiobioside-7-O-β-D-glucoside, quercetin-3-O-β-D-gentiobioside and isorhamnetin-3-O-β-D-gentiobioside respectively. The verage recovery rates (n=9) of three flavonol glycosides compounds were 100.15%(RSD=1.32%), 100.02% (RSD=1.14%), 99.77% (RSD=1.16%), respectively. This method is considered to be simple, accurate, stable, good precision and reproducibility, which is available for the quality control and evaluation of T. terrestris L.
9.Research Progress on Chemical Components and Mechanism of Guava Leaf in Type 2 Diabetes Treatment
Xiangyu GUO ; Tonghua LIU ; Yandi ZHU ; Wen SUN ; Ying DUAN ; Tunhai XU
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(5):1029-1034
Guava leaf (GL) has been widely used as an herbal medicine for diabetes treatment in tropical and sub-tropical regions. GL has the function of drying dampness and fortifying the spleen, clearing heat and resolving toxins. Modern pharmacological study showed that GL contained flavonoid, phenolic acids, terpene, sesquiterpenoids and other chemical compounds, which regulated hyperglycemia. In recent years, the clinical and basic research on GL has received certain progress. This article reviewed effective components of GL, as well as research progress on clini-cal and basic research of type 2 diabetes.
10.Study on Albiflorin Activity in Protection of Schwann Cells in Rats' Sciatic Nerve
Dongchao WANG ; Ying WEI ; Jiaqi GAO ; Yibing LIU ; Lingxia QU ; Yongqiao LIU ; Tunhai XU ; Tonghua LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(5):780-785
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.