2.Recognition of experimental animal model with kidney disease.
Yi-gang WAN ; Yan-ru HUANG ; Wei SUN ; Zhi-min MAO ; Xi-miao SHI ; Jian YAO
China Journal of Chinese Materia Medica 2014;39(21):4075-4081
Animal models with kidney disease are generally divided into two types. One belongs to the models which imitate human kidney disease by the artificial operations, such as anti-glomerular basement membrane antibody nephritis, Heymann nephritis, anti-Thyl. 1 antibody nephritis, BSA nephritis and puromycin nephropathy. The other one pertains to the models which make themselves kidney disease, and appear the pathological characteristics naturally as like as human, such as HIGA mice with IgA nephropathy and NZB/WF1 and MRL/1pr mice with lupus nephritis. In addition,the transgenic animal models with kidney disease can also be established by the modern molecular biologic techniques including gene knockout and siRNA transfection. As for the studies related with kidney disease in pharmacodynamics and pharmacology of Chinese herbal medicine (CHM), it is important to understand deeply the features of each animal model with kidney disease, and select accurately the proper models according to the different experimental objectives, and then, build the special models provided with the combination of disease with syndrome in traditional Chinese medicine (TCM). Therefore,it is the developmental direction for the further study to establish animal models with kidney disease, which should possess the characteristics of syndrome in TCM.
Animals
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Diabetic Nephropathies
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etiology
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Disease Models, Animal
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Humans
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Kidney Diseases
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etiology
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Medicine, Chinese Traditional
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Mice
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Streptozocin
3.TGF - β1 and HSP - 47 expression in pterygium
Hui, YAN ; Ru-Gang, PAN ; Sha-Sha, YAO ; Zhi-Rong, YANG
International Eye Science 2014;(7):1203-1205
AlM:To investigate the expression transforming growth factor-β1 (TGF-β1) and heat shock protein 47 (HSP-47), in pterygium and normal conjunctiva tissues, in order to study the roles of these cytokines played in the pathogenesis in human pterygium.
METHODS:The expression difference of TGF-β1, HSP-47 between human pterygium and normal conjunctive tissues were compared by immuno - histochemistry technique.
RESULTS:The positive expression of TGF-β1, HSP-47 was stronger than in normal conjunctive tissues ( P <0. 05), the TGF-β1 expressed in all layers of pterygium, especially in the squamous epithelium, in the inflammation cells and vascular endothelial cells also expressed. The HSP-47 showed higher expressed in the lamina propria layer of pterygium, and weakly expressed in epithelial layer, no obvious expression in normal conjunctive tissues.
CONCLUSlON:Over-expression of TGF-β1 and HSP-47 in pterygium compared to the normal conjunctiva tissues may play a critical role during the occurrence, development and invasion of the pterygium.
4.Effects and mechanisms of huangkui capsule ameliorating renal fibrosis in diabetic nephropathy rats via inhibiting oxidative stress and p38MAPK signaling pathway activity in kidney.
Zhi-min MAO ; Yi-gang WAN ; Wei SUN ; Hao-li CHEN ; Yan-ru HUANG ; Xi-miao SHI ; Jian YAO
China Journal of Chinese Materia Medica 2014;39(21):4110-4117
OBJECTIVETo demonstrate the effects and mechanisms of Huangkui capsule (HKC) on renal fibrosis in rats with diabetic nephropathy (DN).
METHODRats were randomly divided into 5 groups, the sham-operated group (Sham group, n = 5), the vehicle-given group (Vehicle group, n = 7), the low dose of HKC-treated group (L-HKC group, n = 7), the high dose of HKC-treated group (H-HKC group, n = 7) and the lipoic acid (LA)-treated group (LA group, n = 7). DN models were induced by intraperitoneal injection of streptozotocin (STZ,35 mg x kg(-1)) twice and unilateral nephrectomy. After models were successfully established, the rats in HKC and LA groups were daily administrated with HKC suspensions (0.75, 2 g x kg(-1)) or LA suspensions (60 mg x kg(-1)) respectively, and at the same time, the rats in Vehicle group were daily administrated with distilled water (2 mL) for 8 weeks. All rats were sacrificed at the end of week 8 to collect blood and renal tissues. UAlb, renal function, renal fibrotic morphologic characteristics, as well as oxidative stress (OS)-related markers, the protein expressions of the key signaling molecules in p38 mitogen-activated protein kinase (p38MAPK) signaling pathway, fibrogenic cytokines and inflammatory factors were examined respectively.
RESULTHKC, similar to LA, improved the general state of health, body weight, UAlb, BUN, UA and Alb in DN model rats. Of note, renal fibrosis was ameliorated in HKC groups,especially in H-HKC group which was better than that in LA group. In addition, HKC not only improved the main indexes of OS in the kidney like LA, but also down-regulated the protein expressions of phosphorylated-p38MAPK (p-p38MAPK), transforming growth factor (TGF)-β1 and tumor necrosis factor(TNF)-α in the kidney, whereas, LA only decreased the protein expression of TNF-α in the kidney in DN model rats.
CONCLUSIONHKC, similar to LA, has the actions of anti-OS in vivo. Moreover, HKC could attenuate renal fibrosis by suppressing the activation of p38MAPK signaling pathway and the protein expressions of fibrogenic cytokines and inflammatory factors in the kidney in DN model rats, which is different from LA.
Abelmoschus ; chemistry ; Animals ; Capsules ; Diabetic Nephropathies ; drug therapy ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Fibrosis ; Kidney ; drug effects ; pathology ; MAP Kinase Signaling System ; drug effects ; Male ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors
5.Effects and mechanisms of multi-glycoside of Tripterygium wilfordii improving glomerular inflammatory injury by regulating p38MAPK signaling activation in diabetic nephropathy rats.
Yan-Ru HUANG ; Yi-gang WAN ; Wei SUN ; Zhi-min MAO ; Qing ZHAO ; Xi-miao SHI ; Jian YAO
China Journal of Chinese Materia Medica 2014;39(21):4102-4109
OBJECTIVETo explore the effects and mechanisms of multi-glycoside of Tripterygium wilfordii (GTW) on improving glomerular inflammatory lesion in rats with diabetic nephropathy (DN).
METHODDN model was induced by unilateral nephrectomy and intraperitoneal injection of STZ (35 mg x kg(-1)) twice. The rats were randomly divided into 3 groups, the sham-operated group (Sham group, n = 5), the vehicle-given group (Vehicle group, n = 5 ) and GTW-treated group (GTW group, n = 5). After the model was successfully established, the rats in GTW group were daily oral administrated with GTW suspension (50 mg x kg(-1) x d(-1)), meanwhile, the rats in Vehicle group were daily oral administrated with distilled water (2 mL) for 8 weeks. From the beginning of the administration, all rats were killed 8 weeks later. Blood and renal tissues were collected,and then UAlb, renal function, glomerular morphology characteristics and glomerular macrophages (ED1 + cells) infiltration, as well as the protein expressions of inflammatory cytokines including tumor necrosis factor(TNF)-α and interleukin(IL)-lβ, and the key molecules in p38MAPK signaling pathway including p38 mitogenactivated protein kinase (MAPK), phosphorylated p38 (p-p38MAPK) and transforming growth factor(TGF)-β1 were investigated respectively.
RESULTGTW not only ameliorated the general state of health and body weight,but also attenuated UAlb, glomerulosclerosis, the infiltration of glomerular ED1 + cells and the protein expressions of TNF-α, IL-1β, p-p38MAPK and TGF-β1 in the kidney in DN model rats.
CONCLUSIONBy means of DN model rats, we demonstrated that GTW has the protective effect on renal inflammatory damage in vivo via inhibiting inflammatory cells infiltration and inflammatory cytokines expression. Furthermore, GTW could improve renal inflammatory lesion through down-regulating the expressions of the key signaling molecules in p38MAPK pathway such as p-p38MAPK and TGF-β1 ,and inhibiting the activation of p38MAPK signaling in the kidney.
Animals ; Diabetic Nephropathies ; drug therapy ; Disease Models, Animal ; Glomerulonephritis ; drug therapy ; Glycosides ; pharmacology ; MAP Kinase Signaling System ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; analysis ; Tripterygium ; chemistry ; p38 Mitogen-Activated Protein Kinases ; physiology
6.Regulative mechanisms of oxidative stress in kidney in diabetic nephropathy and interventional effects of Chinese herbal medicine.
Zhi-Min MAO ; Yan-Ru HUANG ; Yi-Gang WAN ; Hao-Li CHEN ; Xi-Miao SHI ; Xian-Jie MENG ; Jian YAO
China Journal of Chinese Materia Medica 2014;39(19):3707-3712
In the development of diabetic nephropathy (DN), reactive oxygen specie (ROS) over much in vivo leads to oxidative stress(OS)-related renal injuries, which are characterized by the structural and functional changes in glomerular and renal tubular cells in morphology. The regulative approaches of OS involve the several signaling pathways, in which, both p38 mitogen-activated protein kinase (MAPK) signaling pathway and adenosine monophosphate-activated protein kinase (AMPK) signaling pathway play the important roles as the target of anti-oxidants. The interventional actions of Chinese herbal compound prescriptions and the extracts of single Chinese herbal medicine (CHM) on OS in the kidney in DN include regulating the balance between ROS and antioxidants, reducing the production of AGEs, inhibiting the expression of growth factors and intervening the activity of signaling pathways.
Animals
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Diabetic Nephropathies
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drug therapy
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metabolism
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Drugs, Chinese Herbal
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administration & dosage
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Humans
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Kidney
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drug effects
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metabolism
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Oxidative Stress
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drug effects
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Signal Transduction
;
drug effects
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Treatment Outcome
7.Effects of electroacupuncture on circadian rhythm of temperature and melatonin in depression rats model induced by chronic stress.
Hai-Jiang YAO ; Hong-Tao SONG ; Yu-Ping MO ; Ting-Ting ZHANG ; Xiang-Bo HAN ; Zhi-Gang LI
Chinese Acupuncture & Moxibustion 2014;34(7):685-689
OBJECTIVETo observe the effects of electroacupuncture (EA) on circadian rhythm of temperature and melatonin (MT) in depression rats model induced by chronic stress, so as to explore the biological mechanism of EA for depression.
METHODSTwenty-four SD rats were randomly divided into a control group, a model group and an EA group, 8 cases in each one. Rats in the control group were treated with normal diet for 21 days without any treatment. In the model and EA group, rat model was established by chronic unpredictable stress combined with solitarily feeding method, and rats in the EA group was treated with EA at "Baihui" (GV 20), "Yintang" (GV 29) 1 h before stress stimulation everyday, 2 Hz in frequency and intensity was favorable with the head of rat slightly shivering. The needles were retained for 20 min, once a day for totally 21 days. After EA treatment, open-field experiment was adopted to observe the behavioral improvement; the rats temperatures were monitored at six time points (2:00, 6:00, 10:00, 14:00, 18:00, 22:00) and orbital blood sampling was collected. The level of serum MT was tested by enzyme linked immunosorbent assay. The circadian rhythm changes of temperature and serum MT in each group were compared.
RESULTSThe numbers of horizontal movement and vertical movement in the model group were obviously lower than those in the control group (both P < 0.05), while those in the EA group were significantly improved compared with those in the model group (both P < 0.01). The circadian rhythm of temperature and MT disappeared in the model group, which was improved into normal level after EA treatment.
CONCLUSIONThe electroacupuncture has regulation effects on circadian rhythm of temperature and melatonin in depression rat model induced by chronic stress.
Animals ; Circadian Rhythm ; Depression ; metabolism ; physiopathology ; therapy ; Electroacupuncture ; Humans ; Male ; Melatonin ; metabolism ; Rats ; Rats, Sprague-Dawley
8.Study on membrane injury mechanism of total alkaloids and berberine from Coptidis Rhizoma on Aeromonas hydrophila.
Dong-fang XUE ; Zong-yao ZOU ; Biao CHEN ; Yan-zhi WANG ; Hao WU ; Xiao-li YE ; Xue-gang LI
China Journal of Chinese Materia Medica 2015;40(9):1787-1792
To explore the antibacterial activity and mechanism of total alkaloids and berberine from Coptidis Rhizoma on Aeromonas hydrophila, and determine the effect of total alkaloids and berberine from Coptidis Rhizoma on minimum inhibitory concentrations, permeability and fluidity of cell membrane, conformation of membrane proteins and virulence factors of A. hydrophila. The results showed that both total alkaloids and berberine from Coptidis Rhizoma had antibacterial activities on A. hydrophila, with minimum inhibitory concentrations of 62.5 and 125 mg · L(-1), respectively. Total alkaloids and berberine from Coptidis Rhizoma could increase the fluidity of membrane, change the conformation of membrane porteins and increase the permeability of bacteria membrane by 24.52% and 19.66%, respectively. Besides, total alkaloids and berberine from Coptidis Rhizoma significantly decreased the hemolysis of exotoxin and the mRNA expressions of aerA and hlyA (P < 0.05, P < 0.01), the secretion of endotoxin and the mRNA expression of LpxC (P < 0.05, P < 0.01). The results suggested that the antibacterial activity of total alkaloids and berberine from Coptidis Rhizoma on A. hydrophila may be related to the bacteria membrane injury. They inhibited the bacterial growth by increasing membrane lipid fluidity and changing conformation of membrane proteins, and reduced the secretion of virulence factors of A. hydrophila to weaken the pathogenicity.
Aeromonas hydrophila
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drug effects
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genetics
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metabolism
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Alkaloids
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pharmacology
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Anti-Bacterial Agents
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pharmacology
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Bacterial Proteins
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genetics
;
metabolism
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Bacterial Toxins
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biosynthesis
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Berberine
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pharmacology
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Cell Membrane
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drug effects
;
genetics
;
metabolism
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Coptis
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chemistry
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Drugs, Chinese Herbal
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pharmacology
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Membrane Fluidity
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drug effects
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Rhizome
;
chemistry
9.Regulative mechanisms of mammalian target of rapamycin signaling pathway in glomerular hypertrophy in diabetic nephropathy and interventional effects of Chinese herbal medicine.
Jing-Jing YANG ; Yan-ru HUANG ; Yi-gang WAN ; Shan-mei SHEN ; Zhi-min MAO ; Wei WU ; Jian YAO
China Journal of Chinese Materia Medica 2015;40(16):3125-3131
Glomerular hypertrophy is the main pathological characteristic in the early stage of diabetic nephropathy (DN), and its regulatory mechanism is closely related to mammalian target of rapamycin (mTOR) signaling pathway activity. mTOR includes mTOR complex 1 (mTORC1) and mTOR complex 2(mTORC2), in which, the upstream pathway of mTORC1 is phosphatidylinositol-3-kinase (PI3K)/serine-threonine kinase(Akt)/adenosine monophosphate activated protein kinase(AMPK), and the representative signaling molecules in the downstream pathway of mTORC1 are 4E-binding proteins(4EBP) and phosphoprotein 70 S6Kinase(p70S6K). Some Chinese herbal extracts could improve cell proliferation via intervening the expressions of the key molecules in the upstream or downstream of PIK/Akt/mTOR signaling pathway in vivo. As for glomerular mesangial cells(MC) and podocyte, mTOR plays an important role in regulating glomerular inherent cells, including adjusting cell cycle, energy metabolism and matrix protein synthesis. Rapamycin, the inhibitor of mTOR, could suppress glomerular inherent cell hypertrophy, cell proliferation, glomerular basement membrane (GBM) thickening and mesangial matrix deposition in model rats with DN. Some Chinese herbal extracts could alleviate glomerular lesions by intervening mTOR signaling pathway activity in renal tissue of DN animal models or in renal inherent cells in vivo and in vitro.
Animals
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Diabetic Nephropathies
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drug therapy
;
enzymology
;
genetics
;
pathology
;
Drugs, Chinese Herbal
;
administration & dosage
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Humans
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Hypertrophy
;
drug therapy
;
enzymology
;
genetics
;
pathology
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Kidney Glomerulus
;
drug effects
;
metabolism
;
pathology
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Signal Transduction
;
drug effects
;
TOR Serine-Threonine Kinases
;
genetics
;
metabolism
10.Neurogenesis by activation of inherent neural stem cells in the rat hippocampus after cerebral infarction.
Bo ZHANG ; Ren-zhi WANG ; Zhi-gang LIAN ; Yang SONG ; Yong YAO
Chinese Medical Sciences Journal 2009;24(1):41-45
OBJECTIVETo investigate the changes of neural stem cells (NSCs) in the rat hippocampus after cerebral infarction (CI) and to evaluate the neurogenesis caused by the activation of NSCs.
METHODSCI models of rats were made and rats were assigned to 6 groups: sham-operated, 1 day, 3 days, 7 days, 14 days, and 28 days after CI. The dynamic expression of bromodeoxyuridine (BrdU), polysialylated neural cell adhesion molecule (PSA-NCAM), glial fibrillary acidic protein (GFAP), and neuronal nuclear antigen (NeuN) were determined by immunohistochemistry and immunofluorescence staining. BrdU was used to mark the proliferated NSCs. PSA-NCAM was used to mark the plasticity of activated NSCs. GFAP and NeuN were used to mark the differentiated NSCs.
RESULTSCompared with the controls, the number of BrdU+ cells in the hippocampus increased significantly at 1 day after CI (P < 0.05), reached peak at 7 days after CI (P < 0.05), decreased but still elevated compared with the controls at 14 days after CI (P < 0.05), and nearly unchanged at 28 days after CI. The number of BrdU+/PSA-NCAM+ cells increased significantly at 7 days after CI (P < 0.05), reached peak at 14 days after CI (P < 0.05), and decreased but still elevated compared with the controls at 28 days after CI (P < 0.05). The number of BrdU+/PSA-NCAM+ cells was equal to 60% of the number of BrdU+ cells in all the same period. The number of BrdU+/NeuN+ cells in the hippocampus increased significantly at 14 days after CI (P < 0.05) and reached peak at 28 day after CI (P < 0.05). The number of BrdU+/GFAP+ cells in the hippocampus nearly unchanged after CI.
CONCLUSIONCI can stimulate the proliferation of inherent NSCs, and most proliferated NSCs may differentiate into neurons and represent neural plasticity.
Adult Stem Cells ; cytology ; metabolism ; Animals ; Bromodeoxyuridine ; metabolism ; Cell Nucleus ; pathology ; Cerebral Infarction ; metabolism ; pathology ; Dentate Gyrus ; cytology ; metabolism ; Glial Fibrillary Acidic Protein ; metabolism ; Hippocampus ; cytology ; metabolism ; Male ; Nerve Tissue Proteins ; metabolism ; Neural Cell Adhesion Molecule L1 ; metabolism ; Neurogenesis ; physiology ; Neurons ; cytology ; metabolism ; Rats ; Rats, Wistar ; Sialic Acids ; metabolism