1.Caffeic acid (CA) protects cerebellar granule neurons (CGNs) from apoptosis induced by neurotoxin 1-methyl-4-phenylpyridnium (MPP~+)
Journal of Peking University(Health Sciences) 2004;0(01):-
Objective: To assess the effects of caffeic acid (CA) on MPP + induced cerebellar granule neurons (CGNs) apoptosis. Methods: CGNs were pretreated with caffeic acid at 55, 110 and 220 ?mol/L for 6 h, then treated with 100 ?mol/L MPP + for 24 h (concentration effect relationship). In addition CGNs were pretreated with caffeic acid at 110 ?mol/L for 0 h, 6 h, 12 h, and 24 h, respectively, then treated with 100 ?mol/L MPP + for 24 h (time response relationship). Besides, after treatment with MPP + for 24 h, CGNs were incubated with caffeic acid at 55, 110 and 220 ?mol/L,respectively. Cell viability was determined by 3 (4,5 dimethylthiazol 2 yl) 2,5 diphenyltetrazolium bromide (MTT) assay and caspase 3 activity was assayed by caspase 3 fluorometric assay kit. Results: MTT assay revealed that caffeic acid significantly inhibited cell viability decrease induced by MPP +, and caspase 3 fluorometric assay showed that caffeic acid efficiently suppressed caspase 3 activation in CGNs induced by MPP +. Conclusion: Caffeic acid (CA) can significantly protect CGNs from apoptosis induced by MPP + and may provide a useful therapeutic strategy for the treatment of Parkinson's disease.
2.Protective Effect of Qiweibaizhu Powder against HRV Infection of Intestinal Mucosal Epithelial Cells of Suckling Mice
Xiao JIANG ; Canrong WU ; Xuefei TIAN
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(05):-
Objective To investigate the protective effect of Qiweibaizhu Powder against HRV infection of intestinal mucosal epithelial cells of suckling mice.Methods The 5-day-old NIH mice were made HRV diarrhea model by oral infection,and randomly divided into six groups:normal group(NG),model group(MG),Qiweibaizhu Powder group(BG),ribavirin group(ZG),suckling mice in each group were instilled corresponding drugs 3 times/d(NG and MG with normal saline) through the mouth.5 d after treatment,suckling mice were killed,small intestine stool was taken to test HRV,and pathological changes in small intestinal mucosa were observed by HE staining.Results Intestinal faeces HRV clearance of BG was significantly better than MG(P
3.Study on Inhibition Effects of Freeze-thawing Leech Extract on HepG2 Cells
Yongliang GUO ; Xuefei TIAN ; Zhu XIAO
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(08):-
Objective To study the inhibition effect of leech extract on HepG2 cells. Methods Human hepatocellular cancer cell line HepG2 were treated with different concentrations of leech extracts which were extracted by method of freeze-thawing with liquid nitrogen and contrasted with that by method of water extracting and ethanol precipitating. The inhibition effects and cell morphous were examined by MTT assay and Acridine orange (AO) fluorescent staining method respectively. Result The 6~15 mg/mL drug concentrations of leech extract by method of freeze-thawing with liquid nitrogen had an obvious inhibition on proliferation of HepG2 cells in a dose-dependent manner, and the effect was better than that by method of water extracting and ethanol precipitating (P
4.Effect of Zijin Capsule on Inflammatory Factors of Bacillary Prostatitis Rats
Qing ZHOU ; Juqiao HE ; Xuefei TIAN
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(09):-
Objective To observe the mechanism of Zijin capsule on treating prostatitis. Methods Bacillary prostatitis rat model was made by injecting E.coil into prostate. The changes of serum TNF-?, IL-2, IL-8, IL-10 and mRNA expression of IL-8 and TNF-? in rat prostate were observed. Results Zijin capsule could reduce the serum TNF-?, IL-8 and prostate TNF-?, IL-8 mRNA expression in rat, increase the level of IL-2 and IL-10. Conclusion Zijin capsule could regulate the immune function of bacillary prostatitis rat model. Participation of immune regulation may be one of the treating mechanisms.
5.Effect of aristolochic acid on human renal cells in vitro
Gongyao TANG ; Xuefei TIAN ; Yipu CHEN
Chinese Journal of Nephrology 1994;0(04):-
0. 05) . (2) AA in the concentrations of 80 and 160?g/ml significantly elevated LDH release rates of HK-2 and hRIFs ( P
6.Relationship between mast cells and renal interstitial fibrosis in interstitial nephritis
Zhao TAN ; Xuefei TIAN ; Hongrui DONG ; Yipu CHEN
Chinese Journal of Nephrology 1997;0(05):-
Objective To explore the potential relationship between tryplase-positive mast cells (MCs) infiltration and renal interstitial fibrosis in acute interstitial nephritis (AIN) and chronic interstitial nephritis (CIN). Methods Renal biopsy specimens from patients with AIN (n=11) and CIN (n=16) were studied and 11 patients in minimal change diseases (MM)were as controls. Histochemistry and immunohistochemistry staining assay were applied to delect the expression of tryptase, proteinase-activated receptor-2 (PAR-2), TGF-?1 and collagen type I (Col I )in the renal tissues. Immunofluo-rescence double-staining assay was used to assess the relationship among MCs, PAR-2-positive cells, and TGF-?1-positive cells in the renal interstitium respectively. Results MCs in AIN and CIN were significantly increased compared with those in controls and were mainly scattered in the fibrotic areas of renal interstitium. The relative immunostaining areas for PAR-2, TGF-?1 in the renal tubular epithelial cells (RTECs) and Col I were significantly larger in AIN and CIN than those in controls respectively (P
7.Preliminary studies on the potential relationship between mast cells and renal interstitial fibrosls in lupus nephritis
Zhao TAN ; Xuefei TIAN ; Yipu CHEN ; Hongrui DONG
Chinese Journal of Rheumatology 2002;0(03):-
Objective To explore the potential relationship between the mast cells (MCs) in renal interstitium and the renal interstitial fibrosis in lupus nephritis (LN). Methods Renal biopsy specimens from patients with types Ⅲ,Ⅳand Vof LN (n=10, respectively), and with minimal change diseases (n=11,as control) were evaluated. Immunohistochemistry staining and immunofluorescence double-staining were used to detect the amount of MCs, the expression of proteinase-activated receptor-2 (PAR-2), transforming growth factor-?1 (TGF-?1) and collagen type I (Col I ) in the renal tissues. Results The amount of MCs in renal interstitium, the positive areas of PAR-2 and TGF-?1 in the renal tubular epithelial cells (RTECs), the amount of PAR-2-positive cells and TGF-?1-positive cells in renal interstitium, and the positive areas of Col I in the renal inter stitium were all higher in three LN groups compared with those in control. Furthermore, among the three LN groups, the above-mentioned parameters were the highest in type Ⅳ and second in type Ⅲ.There were significant positive correlations between the amount of MCs in renal interstitium and the positive areas of PAR-2, TGF-?1 in RTECs as well as the positive areas of Col I in renal interstitium (r=0.513, 0.508, 0.611, respectively, P
8.Screening and analysis of Chinese herbs with liver channel tropism acting on human liver cancer cell differential expressed protein tyrosine kinase
Xuefei TIAN ; Xiaodi HUANG ; Qing ZHOU ; Zhiqi WANG ; Sha TIAN ; Yuan CHEN
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2015;22(3):227-233
Objective To screen the Chinese drugs with liver channel tropism acting on the characteristic differential expressed protein of human liver cancer cell protein tyrosine kinase (PTKs) system by protein chip technology, and to analyze the difference in different Chinese drugs with liver channel tropism in relation to PTKs regulation. Methods Forty BALB/C nude mice were chosen; a piece of subcutaneous tumor mass was implanted into the left lobe of liver parenchyma to reduplicate the orthotopically implanted tumor models. After modeling for 10 days, the tumorigenicity was confirmed, and all the nude mice models were divided into four groups; different Chinese medicine extracts were injected intra-peritoneally into corresponding treatment groups respectively, and the methods of treatment in the 4 groups were as follows: In liver channel tropism drug pair of Huayu Xiaozhen group, rhizoma sparganii and curcuma zedoary with dosage containing crude drug 4.5 g·kg-1·d-1 was given, in liver channel tropism drug pair of Gongdu Sanjie group, the extract of centipede and scorpion with dosage containing crude drug 0.3 g·kg-1·d-1 was intra-peritoneally injected, in spleen channel tropism drug pair control group, astragalus and rhizoma atractylodis macrocephalae with dosage containing crude drug 6.3 g·kg-1·d-1 was given, and in the model group, equal amount of 0.9% normal saline was intra-peritoneally injected. After the drug treatment for 3 weeks, the mice were sacrificed and the liver cancer tissue was obtained; after its total protein was extracted, protein chip technology was applied to screen the Chinese drug pair with liver channel tropism acting on differential expressed PTKs of human liver cancer cells, and the results were analyzed by cluster analysis.Results After the end of the experiment, there were 6 animals in Huayu Xiaozhen drug pair group, 5 in Gongdu Sanjie drug pair group, 5 in control drug pair group and 7 in model group. The protein chip screening results showed that the adjusting fold greater > 1.50 or < 0.67 was defined as the difference with statistical significance. Compared with model group, there were 42 up-regulated expressions of PTKs in Huayu Xiaozhen drug pair group, including 29 receptor tyrosine kinases (RTKs) and 13 non-receptor protein tyrosine kinases (nrPTKs) of which the erythropoietin having adjusting fold over 5.0 produced liver cell receptor B1 (EphB1), epidermal growth factor receptor (ErbB2, ErbB4) etc. 3 RTKs; there were 7 RTKs with adjusting fold 3.0 - 5.0 including EphA1, EphA3, EphA7, fibroblast growth factor receptor 2-α (FGFR2-α), hepatocyte growth factor receptor (HGFR), macrophage colony-stimulating factor receptor (M-CSFR) and vascular endothelial growth factor receptor 2 (VEGFR2), and 2 nrPTKs with adjusting fold 3.0 - 5.0 were non-receptor tyrosine kinase BMX (BMX) and Janus kinase 1 (JAK1). In the Gongdu sanjie drug pair group, there were 23 up-regulated expressions: 15 RTKs and 8 nrPTKs, and 6 down-regulated expressions; the RTKs with adjusting fold 3.0 - 5.0 were ErbB4, M-CSFR and 1 nrPTKs that was megakaryocyte-associated tyrosine kinase (MATK). In the control drug pair group, there were 28 up-regulated and 8 down-regulated expressions. The results of cluster analysis showed that in Huayu Xiaozhen drug pair group, there were 17 differential expressed PTKs in accord with the screen criteria of which 9 were RTKs [receptor tyrosine kinase-like orphan receptor 2 (ROR2), stem cell factor receptor (SCFR), anaplasia lymphoma kinase (ALK), platelet-derived growth factor receptor β (PDGFR-β), insulin-like growth factor-IR receptor (IGF-IR), ErbB2, ErbB3, EphB1 and EphA2], 1 nrPTKs [fps/fes related tyrosine kinase (FER)] and 7 PTKs 3 RTKs (M-CSFR, FGFR2-α, EphA3) and 4 nrPTKs [acetate kinase 1 (ACK1), bruton tyrosine kinase (Btk), non-receptor tyrosine kinase ABL1 (ABL1) and BMX]. In Gongdu Sanjie drug pair group, there were 7 differential expressed PTKs in accord with the screen criteria 5 RTKs (M-CSFR, FGFR1, ROR2, EphB1, ErbB2) and 2 nrPTKs [src-related kinase lacking C-terminal regulation and N-terminal myristylation sites (SRMS), FER]. Conclusions The drug pair of centipede and scorpion with Gongdu Sanjie action possesses a more effective anti-HCC role than the drug pair of rhizoma sparganii and curcuma zedoary with Huayu Xiaozhen action, the mechanism is possibly via the regulation of PTKs signal pathway. The liver channel tropism drug pair of rhizoma sparganii and curcuma with action of promoting blood circulation and removing blood stasis possibly has an independent anti-HCC effective pathway outside the PTKs signal system.
9.Study on Anti-lung Cancer Efficiency of Centipede Extracts in Vitro and Vivo Experiments
Yuan CHEN ; Xiaojia AI ; Zhiqi WANG ; Sha TIAN ; Qing ZHOU ; Gang PEI ; Xuefei TIAN
Chinese Journal of Information on Traditional Chinese Medicine 2016;23(5):61-63
Objective To investigate efficiency of centipede extracts on apoptosis induction, proliferation inhibition to Human A549 cell line and growth suppression of subcutaneous transplanted sarcoma in nude mice. Methods Centipede extracts prepared by enzymolysis and acetone precipitation methods were used to treat human lung cancer A549 cell line. Proliferation inhibition was evaluated by MTT assay and half inhibit concentration (IC50) was calculated. Cell morphological change and apoptosis were detected by flow cytometry and Hoechst stain. The subcutaneous transplanted sarcoma models were prepared with nude mice and randomly divided into model group, control group and centipede extracts group, with 10 mice in each group. Changes of tumor volume, quality and anti-tumor rate were observed.Results In vitro experiment, proliferation of A549 cells was inhibited with dose-dependency and IC50 value was 0.603 mg/mL. The G0/G1 phase of cells was down regulated and G2/M and S phase cells were up-regulated. The apoptotic character cells were been found by Hoechst stain. In vivo experiment, the tumor weight and volume decreased significantly compared with model control group, with statistical significance (P<0.01).Conclusion The centipede extracts shows dose-dependent anti-proliferative effect on A549 cells, which can induce apoptosis by arresting A549 cells at G2/M phase and suppressing growth of subcutaneous transplanted sarcoma of lung cancer in nude mice.
10.Green fluorescent protein expression in vascular endothelial growth factor 165 gene transfected bone marrow mesenchymal stem cells
Yang YANG ; Zhengde HUANG ; Xuefei TIAN ; Desheng ZHOU ; Xinhui LI ; Hua HU ; Yanhong YANG
Chinese Journal of Tissue Engineering Research 2013;(36):6381-6387
BACKGROUND:Vascular endothelial growth factor is a potent angiogenesis and permeability inducible factor. Vascular endothelial growth factor 165 and vascular endothelial growth factor 121 are mainly expressed in vivo, with a strong role of angiogenesis.
OBJECTIVE:To observe the feasibility of vascular endothelial growth factor 165 gene transfected bone marrow mesenchymal stem cel s to differentiate into vascular endothelial cel s.
METHODS:Bone marrow derived mesenchymal stem cel s were isolated and col ected from 50 g Sprague-Dawley rats,and identified by flow cytometry. The plasmid pGLV-EF1a carrying a vascular endothelial growth factor 165 gene was transfected to the mesenchymal stem cel s using lentiviral. Expression of green fluorescent protein was observed under a fluorescence microscope.
RESULTS AND CONCLUSION:After 12 hours of transfection, expression of green fluorescent protein was observed, increased at 48 hours, peaked at 72 hours and gradual y declined thereafter. Results prove that vascular endothelial growth factor 165 gene transfected bone marrow mesenchymal stem cel s have the expression of green fluorescent protein, indicating successful transfection. It is feasible to induce bone marrow mesenchymal stem cel s to differentiate into vascular endothelial cel s.