1.Oxidative damage induced by sodium arsenite in SV-40-immortalized normal uroepithelial cells
Sheng-nan, LIU ; Fei, WANG ; Hui-hui, WANG ; Shu-hua, XI ; Gui-fan, SUN
Chinese Journal of Endemiology 2012;31(1):13-15
ObjectiveTo study the state of oxidative injury induced by sodium arsenite(NaAsO2) in SV-40-immortalized normal uroepithelial (SV-HUC-1 ) cells.Methods SV-HUC-1 cells were exposed to different concentrations of NaAsO2[0(control),1,2,4,8,10 μmol/L] for 24 h,intracellular reactive oxygen species (ROS) was determined by flow cytometry,and the content ofintracellular nitrotyrosine(NT) and the 8-Hydroxydeoxyguanosine (8-OHdG) levels of cell culture medium were detected by enzyme linked immunosorbent assay (ELISA).Results After 24 h treatment,ROS levels(81.76 ± 4.91,95.23 ± 2.17,126.61 ± 17.95,126.74 ± 27.77,114.18 ± 9.65) of SV-HUC-1 cells in the 1,2,4,8,10 μmol/L NaAsO2 exposure groups were significantly higher than those of the control group (69.84 ± 1.28,P < 0.05 or < 0.01 ),ROS levels and exposure dose were positively correlated significantly(r =0.818,P< 0.01); the content of NT in the 10 μmol/L NaAsO2 exposure group[(919.66 ± 206.33) μg/L] was significantly higher than that in the control group[ (238.19 ± 38.28)μg/L,P < 0.01 ],NT content and dye concentrations of arsenic also had dose-response relationship (r =0.617,P < 0.01); after 24 h the cells were treated with arsenic,no significant difference of 8-OHdG content in the culture medium was observed(F =2.127,P > 0.05 ).ConclusionNaAsO2 can cause SV-HUC-1 cell oxidative damage.
3.Determination and evaluation of arsenic speciation in urine of rat exposed to dimethyl arsenic
Lin, ZHANG ; Qing-shan, SUN ; Sheng-nan, LIU ; Ying-li, SONG ; Shu-hua, XI ; Gui-fan, SUN
Chinese Journal of Endemiology 2013;32(6):629-631
Objective To observe the distribution and metabolism of arsenic speciation in urine of rats exposed to different concentrations of dimethylaraenic acid (DMA) through drinking water.Methods Thrity six weaning Wistar rats were randomly divided into normal control,low-dose group and high-dose group,12 rats in each group(6 female and 6 male); average body weight of female rats was (60 ± 5)g,and male rats was (50 ± 5)g.All rats of the 3 groups were given DMA at concentrations of 0,100,200 mg/L,respectively,corresponding to their specific groups through drinking water for 10 weeks.Inorganic arsenic(iAs),monomethylarsenic acid(MMA),DMA and trimethylarsenic compound (TMA) in urine were measured by hydride generation trapping and ultrahypothermia coupled with atomic absorption spectrometry.Results After feeding for 10 weeks,the differences of rat urinary concentrations of iAs,MMA,DMA and TMA between normal control,low-dose group and high-dose group were statistically significant(x2 =25.441,25.942,25.751,17.767,all P< 0.01).Urinary concentrations of iAs,MMA and DMA(2.541,4.383,24.447 mg/L) of low-dose group were significant higher than those of normal control (0.784,0.000,0.743 mg/L,all P < 0.05) ; iAs,MMA,DMA and TMA(3.978,7.186,35.112,4.518 mg/L) of high-dose group were significantly higher than those of normal control(0.784,0.000,0.743,0.000 mg/L,all P < 0.05).The concentrations increased along with increasing doses of DMA concentrations in drinking water(all P < 0.05).Conclusions After rats are exposed to DMA,most of the DMA are excreted in unchanged form in urine and a small portion of DMA is metabolized into TMA.
4.Effect of cariporide on the expression of bcl-2 and bax genes after neck heart transplantation from non heart-beating rats caused by warm ischemia.
Fan-Yan LUO ; Sheng-Xi CHEN ; Lin WANG
Journal of Central South University(Medical Sciences) 2008;33(6):507-511
OBJECTIVE:
To detect the expression of bcl-2 and bax genes after heterotopic heart transplantation in rats that died of warm ischemia, and to explore the effect of cariporide on the protection of the ratos non heart-beating donors.
METHODS:
One hundred and twelve clearing Sprague-Dawley male rats were divided into 7 groups at random (each group contained 16 rats): the control group (Group C), the groups of transplanted hearts after 10, 30, and 45 min of asystolia (Group S10,S30,and S45), and the groups of transplanted hearts after 10,30, and 45 min of asystolia and infused with cariporide(Group SH10,SH30, and SH45).The experimental groups were sacrificed totally by warm ischemia, and heterotopic heart transplantation was processed by the Cuff method. The heart samples of S10,SH10,S30, and SH30 groups were taken at 48 hours after the transplantation, and the heart samples of S45, and SH45 groups were taken just after transplantation. The expression of bcl-2 and bax genes were detected by RT-PCR.
RESULTS:
The death of rats was affirmed when cardiac electric waves vanished after 9~11 minutes of transsection of abdominal aorta. On the RT-PCR test, the expression of bcl-2 gene was the highest and ROD value was maximum in the control group. The expression of bax gene was the lowest and ROD value was minimum in the control group. The ROD value of bcl-2 genes in S10 and S30 groups was less than that in SH10 and SH30 group. The ROD value was just the opposite, and there was stastistical difference (P<0.05).There was no statistical difference between Group S45 and Group SH45 (P>0.05).
CONCLUSION
The model of heteroto-pic neck heart transplantation is a convenient animal model for the cardiac muscle protection. Cariporide can suppress the apoptosis of cardiac muscle cells in rats (within 30 min) after death caused by warm ischemia.
Animals
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Apoptosis
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drug effects
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Guanidines
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pharmacology
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Heart Arrest
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Heart Transplantation
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methods
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Ischemia
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physiopathology
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Male
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Myocardium
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pathology
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Neck
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Proto-Oncogene Proteins c-bcl-2
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biosynthesis
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Random Allocation
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Rats
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Rats, Sprague-Dawley
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Sulfones
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pharmacology
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Tissue Donors
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Transplantation, Heterotopic
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bcl-2-Associated X Protein
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biosynthesis
5.Auricular acupuncture for primary insomnia: a systematic review based on GRADE system.
Hui-Juan TAN ; Ying LAN ; Fu-Sheng WU ; Han-Dan ZHANG ; Li WU ; Xi WU ; Fan-Rong LIANG
Chinese Acupuncture & Moxibustion 2014;34(7):726-730
The clinical efficacy and safety of auricular acupuncture (AA) for treatment of primary insomnia was evaluated. After a comprehensive retrieval in domestic and foreign databases, literatures were strictly screened and Revman 5.2 software was applied to perform a Meta-analysis on eligible randomized controlled trials (RCTs). The evidence quality was assessed with GRADE profiler 3.6 software. As a result, 8 articles were included involving 894 patients. Compared among AA and sham AA, placebo AA, blank control, there was significant difference in Pittsburgh sleep quality index (PSQI) [WMD = -3.48, 95% CI (-3.96, -3.00)], sleep latency LWMD = -10.14, 95% CI (-17.16, -3.12)] and sleep awakening times [WMD = -9.98, 95% CI (-1.10,-0.48)]. Compared between AA and western medication, there was significant difference in PSQI [WMD = -3.62, 95% CI (-4.59, -2.65)]. The evidence quality was moderate in AA vs. sham AA, placebo AA or blank control, while that of the rest was extremely low. No reports of adverse events were described in all studies. In conclusion, for the treatment of primary insomnia, AA could effectively improve sleep quality, but due to the low evidence quality, cautious attitude should be taken on this conclusion, and clinical trials with large sample and high quality were needed in the further.
Acupuncture, Ear
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Humans
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Randomized Controlled Trials as Topic
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Sleep
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Sleep Initiation and Maintenance Disorders
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physiopathology
;
therapy
6.Protection of cadaver lungs of non-heart-beating donor lung transplantation in rats.
Qing-hua HU ; Sheng-xi CHEN ; Fan-yan LUO ; Guo-yong ZHU
Journal of Central South University(Medical Sciences) 2008;33(4):353-358
OBJECTIVE:
To investigate the methods to alleviate lung injury of non-heart-beating donor to attain better structure and function.
METHODS:
Sixty-four Sprague-Dawley rats were randomly divided into 4 groupsú a heart-beating donor group(HBD group), a non-heart-beating donor group without protective measures(NHBD-N group), a non-heart-beating donor group with continuous mechanical ventilation or with topical cooling on cadaver lung (NHBD-V group and NHBD-I group). After the transplantation, lung compliance,pulmonary function,wet/dry ratio of lung and content of energy metabolism were compared among the 4 groups.
RESULTS:
Due to the longer warm ischemic period, NHBD lungs suffered more injuries than HBD lungs. However, compared with NHBD-N group, the wet/dry ratio of the lung in NHBD-V group and NHBD-I group was lower(5.28+/-1.24,4.21+/-0.85,4.14+/-1.33,P<0.01),the lung injury index (14.35+/-3.21,11.28+/-3.26,10.41+/-2.85, P<0.01)and the count of white blood cells(425.60+/-86.47,316.30+/-56.24,295.50+/-70.26, P<0.01) were milder, while the lung compliance and preservation of energetic metabolte were better in the NHBD-V group and the NHBD-I group.
CONCLUSION
Continuous mechanical ventilation or topical cooling may protect the NHBD lung during the warm ischemic period.
Animals
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Cadaver
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Heart Arrest
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metabolism
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physiopathology
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Lung
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metabolism
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physiopathology
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Lung Compliance
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Lung Transplantation
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methods
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Male
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Organ Preservation
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methods
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Random Allocation
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Rats
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Rats, Sprague-Dawley
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Time Factors
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Tissue Donors
7.A study of the relation between interval of the injury and the reaction of neurons, glial cells after experimental brain contusion.
Lian Fang SONG ; Fan Tang MENG ; Xi Yu LIU ; Sheng Hai XIA ; Guang Xun RAO
Journal of Forensic Medicine 2001;17(3):132-136
OBJECTIVE:
To observe the change of c-fos protein(Fos) and nerve growth factor receptor (NGFR) staining in the brain of rat after experimental brain contusion.
METHODS:
Immunohistochemistry of c-fos and NGFR were applied to investigate the brain contusion.
RESULTS:
(1) The expression of Fos protein could be observed at 0.5 h after injury and then increased with the prolonging of time. By 3 h after injury, the positive staining cells could be detected massively not only in and round the wound site but also in other areas of the whole ipsilateral cortex. The stains decreased 6-12 h later and could hardly be detected 1 d after the brain contusion. The control-experiment is negative. (2) NGFR positive staining cells could be found round the wound area 1 d postlesion. At 3 d following injury, a peak of massive positively stained cells appeared both in number and in intensity, showing significant differences compare with that of 1 d after damage (P < 0.01). 5 d later the positive express declined slowly. The express in the control-rat is negative.
CONCLUSION
There is a rule that the expression of Fos and NGFR positive staining changes with time going after brain contusion, which will be of great value in estimation of brain injury time. Detection of Fos can be used for time deduction in earlier period after injury, while NGFR in later period. They are also very important for distinguishing between antemortem or postmortem injury.
Animals
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Brain Concussion/complications*
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Brain Injuries/pathology*
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Female
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Immunohistochemistry
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Male
;
Neuroglia/metabolism*
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Neurons/metabolism*
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Proto-Oncogene Proteins c-fos/metabolism*
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Rats
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Rats, Wistar
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Receptor, Nerve Growth Factor/metabolism*
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Time Factors
8.Influence of ginsenoside Rg1, a panaxatriol saponin from Panax notoginseng, on renal fibrosis in rats with unilateral ureteral obstruction.
Xi-Sheng XIE ; Man YANG ; Heng-Cuang LIU ; Chuan ZUO ; Zi LI ; Yao DENG ; Jun-Ming FAN
Journal of Zhejiang University. Science. B 2008;9(11):885-894
Total saponins of Panax notoginseng (PNS) have been shown to ameliorate renal interstitial fibrosis. Ginsenoside Rg1, a panaxatriol saponin, is one of the major active molecules from PNS. The present study was undertaken to investigate the effect of ginsenoside Rg1 on renal fibrosis in rats with unilateral ureteral obstruction (UUO). The rats were randomly divided into 3 groups: sham-operation (n=15), UUO (n=15) and UUO with ginsenoside Rg1 treatment (n=15, 50 mg per kg body weight, intraperitoneally (i.p.) injected). The rats were sacrificed on Days 7 and 14 after the surgery. Histological examination demonstrated that ginsenoside Rg1 significantly inhibited interstitial fibrosis including tubular injury as well as collagen deposition. alpha-smooth muscle actin (alpha-SMA) and E-cadherin are two markers of tubular epithelial-myofibroblast transition (TEMT). Interestingly, ginsenoside Rg1 notably decreased alpha-SMA expression and simultaneously enhanced E-cadherin expression. The messenger RNA (mRNA) of transforming growth factor-beta1 (TGF-beta1), a key mediator to regulate TEMT, in the obstructed kidney increased dramatically, but was found to decrease significantly after administration of ginsenoside Rg1. Further study showed that ginsenoside Rg1 considerably decreased the levels of both active TGF-beta1 and phosphorylated Smad2 (pSmad2). Moreover, ginsenoside Rg1 substantially suppressed the expression of thrombospondin-1 (TSP-1), a cytokine which can promote the transcription of TGF-beta1 mRNA and the activation of latent TGF-beta1. These results suggest that ginsenoside Rg1 inhibits renal interstitial fibrosis in rats with UUO. The mechanism might be partly related to the blocking of TEMT via suppressing the expression of TSP-1.
Actins
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biosynthesis
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Animals
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Cadherins
;
biosynthesis
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Collagen Type I
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genetics
;
metabolism
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Fibronectins
;
genetics
;
metabolism
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Ginsenosides
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pharmacology
;
Immunohistochemistry
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Male
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Nephritis, Interstitial
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drug therapy
;
genetics
;
metabolism
;
pathology
;
Panax notoginseng
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chemistry
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RNA, Messenger
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biosynthesis
;
genetics
;
Random Allocation
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Rats
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Rats, Sprague-Dawley
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Reverse Transcriptase Polymerase Chain Reaction
;
Smad2 Protein
;
biosynthesis
;
Thrombospondin 1
;
biosynthesis
;
genetics
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Transforming Growth Factor beta1
;
biosynthesis
;
genetics
;
Ureteral Obstruction
;
metabolism
;
pathology
9.Ginsenoside Rb1, a panoxadiol saponin against oxidative damage and renal interstitial fibrosis in rats with unilateral ureteral obstruction.
Xi-sheng XIE ; Heng-chuan LIU ; Man YANG ; Chuan ZUO ; Yao DENG ; Jun-ming FAN
Chinese journal of integrative medicine 2009;15(2):133-140
OBJECTIVETo investigate the possible protective effect and mechanism of ginsenoside Rb1 against oxidative damage and renal interstitial fibrosis on rats with unilateral ureteral obstruction (UUO).
METHODSIn total, 80 male rats were randomly divided into 4 groups, 20 in each group: the sham operated group (SOR), UUO group, UUO with ginsenoside Rb1 treatment group (treated with intraperitoneal injection of 50 mg/ kg daily) and UUO with Losartan treatment group (as the positive control, treated with 20 mg/kg by gastrogavage per day). The rats were randomly sacrificed on day 3, 7 and 14 after surgery, respectively. The histopathologic changes of renal interstitial tissues were observed with Masson staining. The mRNA of transforming growth factor beta 1 (TGF-beta 1), collagen I and fibronectin were reversed transcribed and quantified by Real-time PCR. Enzyme-linked immunosorbent assay was used to quantitatively detect TGF-beta 1 and 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels. P47phox protein expression was assessed by immunohistochemistry and Western blot analysis.
RESULTSIn the UUO model, the obstructed kidney showed typical features of progressive renal tubulointerstitial fibrosis, and the levels of TGF-beta1, collagen I and fibronectin increased (P<0.05). As compared with the UUO group, ginsennoside Rb1 significantly inhibited the interstitial fibrosis including tubular injury and collagen deposition, and decreased the levels of TGF-beta1 (P<0.05). Ginsenoside Rb1 also inhibited the heme oxygenase (HO-1) and 8-OHdG, two markers of oxidative stress (P<0.05). Moreover, ginsenoside Rb1 suppressed the expression of p47phox, a subunit of nicotinamide adeninedinucleotide phosphate (NADPH) oxidase (P<0.05).
CONCLUSIONGinsenoside Rb1 can obviously inhibit renal interstitial fibrosis in rats with UUO, its mechanism possibly via against the oxidative damage and suppressing TGF-beta1 expression.
Animals ; Deoxyguanosine ; analogs & derivatives ; urine ; Drug Evaluation, Preclinical ; Fibrosis ; genetics ; metabolism ; prevention & control ; Gene Expression Regulation ; drug effects ; Ginsenosides ; therapeutic use ; Heme Oxygenase (Decyclizing) ; metabolism ; Kidney ; drug effects ; metabolism ; pathology ; Kidney Diseases ; etiology ; genetics ; pathology ; prevention & control ; Male ; Models, Biological ; NADPH Oxidases ; genetics ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Saponins ; therapeutic use ; Transforming Growth Factor beta1 ; genetics ; metabolism ; Ureteral Obstruction ; complications ; drug therapy ; genetics ; metabolism
10.Ginsenoside R(g1) inhibit transdifferentiation in rat renal tubular epethelial cells induced by TGF-beta1.
Xi-sheng XIE ; Heng-chuan LIU ; Hui-juan LI ; Jun-ming FAN
China Journal of Chinese Materia Medica 2008;33(17):2136-2141
OBJECTIVETo investigate the effects of ginsenoside R(g1) on the transdifferentiation of rat renal tubular epethelial cells induced by transforming growth factor-beta1, (TGF-beta1).
METHODCultured normal rat renal tubular epethelial cells (NRK-52E) were divided into control group, TGF-beta1-induced group and treated with ginsenoside R(g1) at different concentration (10, 20, 40 mg x L(-1)) group. The morphology of tubular epithelial-myofibroblast transdifferentiation induced by TGF-beta1 was observed through light microscope. alpha-SMA and E-cadherin protein expression were assessed by immunohistochemistry and western blot analyses. alpha-SMA, collagen I and and fibronectin gene expression were assessed by real-time quantitative chain reaction. Enzyme-linked immunosorbent assay was used to quantitatively detect collagen I and fibronectin in the supernatant.
RESULT10 mg x L(-1) TGF-beta1 could induce the transdifferentiation of tubular epithelial myofibroblast, showing fibroblast-like in morphology, with significantly enhanced expression of alpha-SMA, depressed expression of E-cadherin and increased secretion of fibronectin and collagen I (P < 0.05). Compared to TGF-beta1-induced group, ginsenoside R(g1) partly abrogated the alpha-SMA expression and E-cadherin depression triggered by TGF-beta1 in tubular epithelial cells in a dose-dependent manner (P < 0.05). Meanhile, ginsenoside R(g1) blocked morphologic transformation of tubular epithelial cells and decreased levels of collagen I and fibronectin (P < 0.05).
CONCLUSIONGinsenoside R(g1) could inhibit TGF-beta1 induced the tubular epithelial-myofibroblast transdifferentiation and decreased levels of collagen I and fibronectin in NRK52E.
Animals ; Cadherins ; genetics ; metabolism ; Cell Line ; Cell Transdifferentiation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; cytology ; drug effects ; Gene Expression ; drug effects ; Ginsenosides ; pharmacology ; Kidney Tubules ; cytology ; drug effects ; Panax ; chemistry ; Rats ; Transforming Growth Factor beta1 ; pharmacology