1.Effects of Acidic Fibroblast Growth Factor on Hippocampal Astrocytes Injury Induced by Gentamicin:Role of Mitogen-Activated Protein Kinase Signal Pathway
Li SHEN ; Juen HUANG ; Taiping HUANG ; Hui SHEN ; Xiaokun LI
Chinese Journal of Rehabilitation Theory and Practice 2016;22(3):270-273
Objective To explore the mechanism of the protection of acidic fibroblast growth factor (aFGF) for hippocampal astrocytes from injury induced by gentamicin. Methods Hippocampal astrocytes were isolated from newborn (24 hours) Sprague-Dawley rats, puri-fied, and identified with glial fibrillary acidic protein (GFAP) immunofluorescence. The third generations were cultured for 3 days and divid-ed into 3 groups:control group was cultured routinely, injury group was cultured with 2.0 g/L gentamicin for 24 hours, and protection group was cultured with 4.25μg/L aFGF for 24 hours and then cultured with 2.0 g/L gentamicin for 24 hours. Western blotting was adopted to de-tect the expressions of P38, extracellular signal-regulated kinase (ERK)1/2 and c-Jun N-terminal kinase (JNK)1/2. Results Hippocampal as-trocytes were culturated successfully with the purity above 95%. The ERK1 increased in the injury group compared with the control group (P<0.05). Compared with the injury group, the p38 increased (P<0.05) and the ERK1 decreased (P<0.05) in the protection group. There was no significant difference among others (P>0.05). Conclusion The mitogen-activated protein kinase signal pathway, especially P38 and ERK1, may associate with the protection of aFGF for hippocampal astrocytes from injury induced by gentamicin.
2.Protective effects of bFGF on rats’renal tubular epithelial cells damaged by gentamicin in vitro
Juen HUANG ; Quanyong HUANG ; Weiping CHEN ; Zuding LIANG
Chinese Pharmacological Bulletin 1987;0(02):-
Aim To study the protective effects of bFGF on rats’ renal tubular epithelial cells injured by gentamicin in vitro.Methods After renal cortexes were grinded,filtered and digested with 0.25% trypsin,renal tubular epithelial cells were cultured.The cell types of epithelial cells were identified by cytokeratin 18 immunocytochemistry.The injury models of epithelial cells of renal tubules were established by gentamicin and then the protective roles of bFGF on injured renal tubular epithelial cells were observed. Results ①By observing the morphology of the epithelial cells and identifying the cell types with cytokeratin 18 antibody,comparing GM group with control group,CAT,Na+,K+-ATP enzyme activity decreased,while NAG activity and MDA level increased(P0.05),but the differences of MDA was still significant(P
3.Protective effect of modified acidic fibroblast growth factor on renal ischemia/reperfusion injury of rats
Wenjuan HU ; Juen HUANG ; Xiaokun LI ; Jian XIAO
Chinese Pharmacological Bulletin 1987;0(02):-
Aim To study the protective effect of modified acidic fibroblast growth factor (MaFGF) on renal ischemia/reperfusion injury of rats. Methods 32 Wistar rats were divided randomly into four groups;false operative control group, model control group, 8 ?g?kg-1 of MaFGF and 16 ?g?kg-1 of MaFGF groups with 8 animals in each group. Renal pathological changes were observed by light and electron microscope. Results As shown in pathology, compared with model control group, MaFGF group could significantly reduce edema, number of leukocytes appearing in the interstitium ,distention and destroy of renal tissues. Under EM, the ultrostructure(including microvilli, mitochondria and lysosomes etc.) of the tubular epithelial cells of MaFGF groups appeared lighter in injuries or even near normal. Conclusion MaFGF has a role in attenuation of renal damage or failure after ischemia-reperfusion injury of the kidney.
4.Experimental research on the role of acidic fibroblast growth factor in the survival and proliferation of renal tubular epithelial cells in vitro
Limin LIU ; Huagang LIU ; Juen HUANG ; Xiaokun LI ; Jian XIAO
Chinese Pharmacological Bulletin 1986;0(06):-
Aim To explore effects of acidic fibroblast growth factor(aFGF)on the growth of renal tubular epithelial cells cultured in vitro. Methods Renal tubular epithelial cells in rats were gained through digestion by pamcreatin. aFGF was added into cultures of renal tubular epithelial cells, as experimental groups. The number of cells was counted and the shape of cells was observed at 12,24,48 and 72 h. Results Renal tubular epithelial cells were successfully obtained from kidneys. After treated for 72 h,the renal tubular epithelial cells showed different proliferation in experimental and control groups. There was no obvious difference at 12 h, but there was statistical difference between the two groups at 24 h,48h and 72 h(P
5.AN IMMUNOELECTRON MICROSCOPIC STUDY ON THE SYNAPTIC RELATIONSHIP BETWEEN DOPAMINERGIC AND GABAergic NEURONS IN THE BASOLATERALNUCLEUS OF THE RAT AMYGDALA
Xiaoyun QIN ; Juen HUANG ; Naixiang LUO ; Bojun WANG ; Changwen JIANG
Acta Anatomica Sinica 1957;0(04):-
Objective To explore the relationship between dopaminergic axon terminal and GABAergic neurons and explore the neuroanatomic mechanism of their effects in schizophrenia. Methods The co-location and the association of dopaminergic axon terminal and GABAergic neurons in the basolateral nucleus (BL) of rat amygdala were examined by using double labeling immunoelectron microscopic techniques. Dopaminergic axon terminal and GABAergic neurons were labeled with the antidopamine (anti-DA) and the anti-glutamic acid decarboxylase (anti-GAD) antibodies respectively. Results 43% of the DA-input synapses was observed to relate directly or indirectly to GAD-immunoreactive(IR) dendritic structures(DA/GAD), which includes single (38%), convergent (30%), serial (20%), and axoaxonic (12%) contact types.And 57% of the DA-input synapses was found to associate with unlabeled elements (DA/UE), which includes unlabeled perikarya (10%), dendrites (82%) and axons (8%). All of the synapses that show DA-IR terminals profiles were found to be symmetric (inhibitory) synapses.Conclusion These results suggest that the dopaminergic system in the BL of rat amygdala controls the mediations of the GABAergic interneurons via symmetric synapses. In addition, the dopaminergic axon terminal associates with the glutamatergic projection neurons and exerts influence on its activity.
6.Protective effect of basic fibroblast growth factor on renal tubular epithelial cell toxicity induced by gentamicin
Rongshi HUANG ; Juen HUANG ; Yanhui CEN ; Guozhen HE ; Feilan ZHAO ; Rong FAN
Chinese Journal of Tissue Engineering Research 2010;14(2):240-244
BACKGROUND: In vivo experiments have confirmed that fibroblast growth factor can effectively protect gentamicin-induced renal tubular epithelial cell injury, but the effect on the in vitro cultured cells is still rare. OBJECTIVE: To explore the mechanisms of basic fibroblast growth factor (bFGF) at different concentrations on preventing nephrotoxity mediated by genamicin on the primarily cultivated renal tubular epithelial cell models. METHODS: By use of enzyme and mesh screening, renal tubular epithelial cells were isolated from Kunming mice and purified, adjusting the cell concentration of 1×10~8/L, then cell suspension was moved to a 96-well culture plate and divided into different groups for culture: blank control group: normal culture; gentamicin group: 10, 30, 50 μL/hola (ie, 400, 1 200, 2 000 U/holes)arerecorded as G1, G2, G3; bFGF group: 20, 50, 80 μL/hole (ie, 90,225, 360 ng/hole) are recorded as B1, B2, B3; gentamicin plus bFGF intervention group: after adding bFGF 12 hours, then added gentamicin 12 hours, assigned into 9 dose subgroups, namely, G1B1, G1B2, G1B3, G2B1, G2B2, G2B3, G3B1, G3B2, G3B3, each subgroup contained four-hole complex. Cell morphology and quantity was observed. RESULTS AND CONCLUSION: Gentamicin showed a dose-dependent effect on the renal tubular epithelial cell injury, epithelial cells in the medium and high concentration groups exhibited shrinkage, rounded, swelling, poor adhesion, severely damaged cytoplasm and structural disorder. In the low concentration group, the number change of cells was not obvious, and fibroblasts began to appear; In the bFGF groups, cells were full, exhibited strong refraction, the cell number increased significantly, these manifestations were significant in 50 μL/hole concentration, and there was no significant difference compared with 80 μL/hore concentration; in case of gentamicin plus bFGF intervention, cells with low concentrations of gentamicin had no obvious damage to cells, which increased, the damaged cells collapse was reduced in the group of low concentration of gentamicin, cell shrinkage and poor adhesion were slightly relieved, high concentrations of bFGF intervention could yield to good cell morphology, but high concentrations of gentamicin caused cell swelling and necrosis of injury, which could not be improved by any concentrations of bFGF intervention. 50 μL/hole bFGF has antagonistic effect on the nephrotoxicity mediated by medium and low concentrations of gentamicin, but has no protection on high concentration of gentamicin-induced nephrotoxicity.
8.Protective effects of aFGF on gentamicin-induced injuries of rats'renal tubular epithelial cells in vitro
Juen HUANG ; Huijie WANG ; Huagang LIU ; Haiyan LIU ; Xiaokun LI ; Jian XIAO
Chinese Pharmacological Bulletin 2003;0(11):-
Aim To study protective effects of aFGF on rats' renal tubular epithelial cells injured by gentamicin in vitro.Methods After renal cortice were grounded,filtered and digested with 0.25%trypsin,renal tubular epithelial cells were cultured.The injury models of epithelial cells of renal tubules were established by gentamicin and then the protective roles of aFGF in injured renal tubular epithelial cells were observed.Results ① By observing the morphology of the epithelial cells and comparing GM group with control group,CAT、SOD、Na~+,K~+-ATP enzyme activities decreased,while NAG activity and MDA level increased(P
9.Protective effects of (-)-epigallocatechin-3-gallate on renal ischemia-reperfusion injury
Gang LIANG ; Juen HIANG ; Xiao ZHANG ; Xingguang LIAO ; Ying XU ; Renbin HUANG ;
Chinese Pharmacological Bulletin 1987;0(01):-
AIM To study the protective effects of ( ) epigallocatechin 3 gallate (EGCG) against renal ischmia reperfusion injury. METHODS The renal ischemia reperfusion injury model in rat was made by means of ligating bilateral renal arteries for 60 min and then reperfusion. 10 mg?kg -1 and 40 mg?kg -1 EGCG were given by intravenation before and after operation respectively. Spectrophotometric assay were used to measure the contents of serum creatinine(Scr), blood urea nitrogen (Bun) in serum and the activity of superoxide dismutase (SOD) and Ca 2+ ATPase ,the contents of malondialdehyde (MDA), reactive oxygen species (ROS) in renal hemogenate from rats. Renal pathologic changes were also observed. RESULTS Compared with model control group , 40 mg?kg -1 EGCG group was found of significant inhibition in changes of Scr, Bun,MDA,ROS and significant increases in activity of SOD and Ca 2+ ATPase.The renal morphological injury of EGCG groups were slighter than that of model control group. CONCLUSION EGCG has protective effects aganist renal ischemia reperefusion injury via its antioxidant activity and intracellular caclcium reduction.