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1.
Chinese Journal of Tissue Engineering Research ; (53): 2033-2038, 2018.
Article in Chinese | WPRIM | ID: wpr-698654

ABSTRACT

BACKGROUND: Currently, mesenchymal stem cells have been widely explored and applied in scientific research field, and many studies suggest that the underlying mechanism of mesenchymal stem cells mainly relies on its exosomes. OBJECTIVE: To isolate and identify human adipose-derived stem cells and its exosomes, and to identify their biological characteristics. METHODS: Human adipose tissue was digested with collagenase l, and adipose-derived stem cells were isolated and purified. Immunophenotype, osteogenic and adipogenic abilities of adipose-derived stem cells were identified. Exosomes were isolated by using ultrafiltration method. Morphology of exosomes was observed by Nanosight and electron microscope. Expression of proteins in exosomes was detected by antibody array method. RESULTS AND CONCLUSION: Adipose-derived stem cells exhibited long spindle-like or fibroblast-like appearance, expressed CD73, CD44, CD90, CD105 and had the potential to differentiate into many tissues, including bone and adipose tissues. The exosomes had the similar size, with the diameter of 30-150 nm. They possessed many proteins including FLOT1, ICAM, ALIX, CD81, CD63, ANXA5, TSG101, and so on. Findings from the present study indicate the successful isolation of exosomes from human adipose-derived stem cells.

2.
Chinese Journal of Hepatology ; (12): 826-830, 2014.
Article in Chinese | WPRIM | ID: wpr-337097

ABSTRACT

<p><b>OBJECTIVE</b>To study expression of regucalcin (RGN) and prohibitin (PHB) genes in cirrhotic rat liver and to investigate the related effects of compound glutathione inosine injection (CGII) intervention.</p><p><b>METHODS</b>Forty male Wistar rats were randomly divided into a control group (n=12) and a model group (n=28).The model was established by injecting sterile porcine serum (0.5 mL) into the rat abdominal cavity, twice weekly for 8 consecutive weeks; the control group rats were treated with physiological saline injection (0.5 mL) into the abdominal cavity with the same frequency and time span. During the modeling period, four rats from the model group were randomly selected at different time points to examine changes in liver pathology. Upon pathology confirmation of liver cirrhosis, the porcine serum injection was terminated. The remaining 24 rats in the model group were randomly divided into a fibrosis group and a CGII treatment group.The CGII group received CGII (intramuscular injection of 0.018 mL 100g-1 body weight) once a day for 6 continuous weeks; the fibrosis rats were treated with the same dosage of physiological saline with the same frequency and time span.Liver tissue morphology was examined by both hematoxylin-eosin and Masson's staining. RGN and PHB expression at the mRNA and protein levels in liver tissues were detected by real time RT-PCR and immunohistochemical staining, respectively.</p><p><b>RESULTS</b>Both the mRNA and protein expression levels of RGN and PHB were significantly lower in the liver tissues of the fibrosis group than in the control group.CGII intervention led to significant alleviation of the liver fibrosis severity; moreover, the mRNA and protein expression levels of RGN and PHB were significantly higher than those in the fibrosis group.</p><p><b>CONCLUSION</b>Down-regulation of regucalcin and prohibitin gene expression might contribute to the pathogenesis of liver cirrhosis.</p>


Subject(s)
Animals , Male , Rats , Calcium-Binding Proteins , Genetics , Down-Regulation , Gene Expression , Glutathione , Toxicity , Inosine , Intracellular Signaling Peptides and Proteins , Genetics , Liver Cirrhosis , Genetics , Rats, Wistar , Repressor Proteins , Genetics
3.
Chinese Journal of Hepatology ; (12): 589-592, 2012.
Article in Chinese | WPRIM | ID: wpr-296843

ABSTRACT

To investigate the efficacy of interferon alpha 2 b plus ribavirin combination therapy in sixty-two patients with chronic hepatitis c (CHC) infection originating from a single blood donor. The 62 patients who developed CHC following blood transfusion from a known single infected donor were treated with interferon and ribavirin combination therapy for 48 weeks and followed-up for 96 weeks. The therapy regimen consisted of subcutaneous administration of 3-500 MIU interferon alpha 2 b every other day and daily oral administration of 0.6-1.0 g of ribavirin. Patients were monitored during treatment and in follow-up for sustained virological response (SVR), early virology response (EVR), treatment end virology response (ETVR), biochemical response of withdrawals, and side effects. The SVR rate was 83.9% (52/62). The EVR rate was 95.2% (59/62). The ETVR rate was 87.1% (54/62). The biochemical response rate after withdrawal of treatment was 100.0%. Eight patients developed mildly abnormal thyroid function as a result of the interferon therapy, but all were able to complete the antiviral treatment regimen under the care of endocrinologists. Younger age, relatively short course of disease, low viral load, and better compliance, but not sex, were correlated to curative effect of the combination therapy. Interferon alpha 2 b plus ribavirin combination therapy had a significant curative effect on a group of 62 CHC patients originating from a single case, with 52 of the patients showing SVR out to 96 weeks after therapy. Antiviral treatment is recommended for hepatitis C virus-positive patients to eradicate the virus and prevent disease progression.


Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Middle Aged , Young Adult , Antiviral Agents , Therapeutic Uses , Blood Donors , Drug Therapy, Combination , Follow-Up Studies , Hepacivirus , Genetics , Hepatitis C, Chronic , Drug Therapy , Virology , Interferon-alpha , Therapeutic Uses , RNA, Viral , Blood , Recombinant Proteins , Therapeutic Uses , Ribavirin , Therapeutic Uses , Transfusion Reaction , Treatment Outcome , Viral Load
4.
Chinese Journal of Hepatology ; (12): 497-502, 2012.
Article in Chinese | WPRIM | ID: wpr-261966

ABSTRACT

<p><b>OBJECTIVE</b>To study the protective effects of blueberry against rat immune hepatic fibrosis, specifically through the expression of hepatic cytochrome P450 2E1.</p><p><b>METHOD</b>Fifty Wistar rats were randomly divided into five study groups (n = 10 each): Group A: normal control group, Group B: hepatic fibrosis model group, Group C: preventive group administered blueberry juice, Group D: preventive group administered Fu-Fang-Bie-Jia-Ruan-Gan tablet, and Group E: preventive group administered a combination of blueberry juice and Fu-Fang-Bie-Jia-Ruan-Gan tablet. The hepatic fibrosis model was established by intraperitoneal injection of porcine serum once daily for 12 weeks. Simultaneously, rats in preventive groups (Groups C-E) were perfused with blueberry juice or Fu-Fang-Bie-Jia-Ruan-Gan tablet or combinations of blueberry juice and Fu-Fang-Bie-Jia-Ruan-Gan tablet, respectively, for 12 weeks. The normal control group was perfused with saline for 12 weeks. All animals were sacrificed at the end of the 12 weeks, and serum levels of alanine aminotransferase (ALT) were measured and activities of superoxide dismutase (SOD), malondialdehyde (MDA), and hydroxyproline (Hyp) in liver homogenates were determined. Pathology of hepatic fibrosis was evaluated by hematoxylin-eosin (HE) and Masson staining. Expression of CYP2E1 was detected by real-time RT-PCR, immunohistochemical techniques, and Western blotting.</p><p><b>RESULTS</b>Serum ALT levels were not significantly different in the control and treatment groups (F=4.056, P more than 0.05): A: 37.87+/-4.53 U/L, B: 49.23+/-9.81 U/L, C: 39.94+/-6.32 U/L, D: 40.50+/-5.70 U/L, and E: 38.24+/-8.43 U/L. Compared with Group B, the pathological stages of hepatic fibrosis were significantly reduced in the prevention groups (C-E) (F=95.097, P less than 0.05). Hyp and MDA in liver homogenates of groups C-E were significantly lower than those of Group B (Hyp: C: 472.68+/-44.14 mug/g, D: 416.12+/-39.38 mug/g, E: 429.51+/-55.14 mug/g vs. B: 603.16+/-68.92 mug/g, F=39.315, P less than 0.05; MDA: C: 0.83+/-0.06 nmol/mg, D: 0.96+/-0.08 nmol/mg, E: 0.85+/-0.06 nmol/mg vs. B: 1.24+/-0.15 nmol/mg, F=46.376, P less than 0.05). In contrast, SOD activities in Group C-E were significantly higher than those in Group B (C: 2.47+/-0.38 U/mg, D: 1.95+/-0.45 U/mg, E: 2.16+/-0.23 U/mg vs. B: 1.56+/-0.41 U/mg, F=25.557, P less than 0.05). Compared with Group A, the mRNA and protein expressions of CYP2E1 were increased in groups B-E, however the differences did not reach statistical significance (mRNA: F=0.897, protein: F=0.492, both P more than 0.05). The mRNA and protein expressions of CYP2E1 in groups C-E were lower than those of Group B, however the differences did not reach statistical significance (mRNA: F=0.897, protein: F=0.492, P more than 0.05).</p><p><b>CONCLUSION</b>Blueberry exhibits certain protective effects against porcine serum-induced hepatic fibrosis in rats. The expression of hepatic cytochrome P450 2E1 in rats with immune hepatic fibrosis is not significantly different from the normal rats. Blueberry has no effect on the expression of hepatic cytochrome P4502E1.</p>


Subject(s)
Animals , Male , Rats , Blueberry Plants , Cytochrome P-450 CYP2E1 , Metabolism , Drugs, Chinese Herbal , Pharmacology , Fruit , Immune System Diseases , Metabolism , Liver Cirrhosis, Experimental , Metabolism , Pathology , Malondialdehyde , Metabolism , Plant Extracts , Pharmacology , Rats, Wistar , Superoxide Dismutase , Metabolism
5.
Chinese Journal of Hepatology ; (12): 349-351, 2011.
Article in Chinese | WPRIM | ID: wpr-290596

ABSTRACT

<p><b>OBJECTIVE</b>To find out clinical characteristics and natural history of post transfusion HCV infection.</p><p><b>METHODS</b>83 subjects who have received the blood from a same blood donor from January 1998 to July 2002 were investigated by the method combining cross-sectional study with retrospective study. HCV-antibody, HCV RNA, liver function, abdomen B-ultrasound and Fibroscan were detected.</p><p><b>RESULTS</b>The HCV-antibody were all positive. The HCV RNA of 56 out of the 83 cases were positive. The chronicity rates of hepatis C were 76.3% (29/38) in male patients and 60.00% (27/45) in female patients respectively, without significant difference (X² = 2.99, P = 0.11). The average age of the HCV RNA positive patients was (36.54 ± 14.37) years old. The average age of the HCV RNA negative patients was (27.43 ± 12.51) years old. A significant difference (T = -2.41, P = 0.018) existed between. The HCV genotype was type1b. Among the HCV RNA positive patients,10 cases were with mild asthenia, anorexia and abdominal distention, 9 cases with increased serum ALT, 12 cases.with chronic hepatitis and 1 case was diagnosed with decompensated liver cirrhosis.</p><p><b>CONCLUSION</b>The clinical manifestations of HCV infection are occult and chronic. The chronicity rate is related to gender and the age when infection was caught.</p>


Subject(s)
Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Young Adult , Age Factors , Blood Donors , Cross-Sectional Studies , Follow-Up Studies , Hepacivirus , Hepatitis C , Virology , Hepatitis C, Chronic , Virology , Retrospective Studies , Sex Factors , Transfusion Reaction
6.
Chinese Journal of Hepatology ; (12): 656-660, 2010.
Article in Chinese | WPRIM | ID: wpr-360873

ABSTRACT

<p><b>OBJECTIVE</b>To study the protective effect of Blueberry against rat hepatic fibrosis and the effect of Blueberry on HO-1 expression patterns.</p><p><b>METHODS</b>A total of 45 SD rats were randomly divided into five groups namely control group (group A), model group (group B), blueberry group (group C), Dan-shao-hua-xian (DSHX) capsule group (group D) and blueberry +Dan-shao-hua-xian group (group E). Fibrous liver models in rats were induced by subcutaneous injection of CCl4 and high-lipid/low-protein diet for 8 weeks except the control group which accepted saline alone. The level of alanine aminotransferase (ALT) in serum was examined. The activities of superoxide dismutase (SOD) and malondialdehyde (MDA) in liver homogenates were determined. by the xanthine oxidase method and the thiobarbituric acid method. The pathology of hepatic fibrosis was evaluated by hematoxylin and eosin (H and E) staining. The Expression of HO-1 was detected by real-time RT-PCR, immunohistochemical techniques and western blotting.</p><p><b>RESULTS</b>Serum ALT levels in every prevention group was lower than the group B [(149.44+/-16.51), (136.88+/-10.07), (127.38+/-11.03) vs (203.25+/-31.62) U/L, F = 92.498, P < 0.05], the SOD of liver homogenate in prevention group was significantly higher and the MDA was lower compared with the group B [SOD: (1.36+/-0.09), (1.42+/-0.13), (1.50+/-0.15) vs (1.08+/-0.19) U/mg, F = 13.671, P < 0.05; MDA: (0.294+/-0.026), (0.285+/-0.025), (0.284+/-0.028) vs (0.335+/-0.056) nmol/mg, F = 20.809, P < 0.05]. The pathological stages of hepatic fibrosis were all significantly reduced in prevention group (Chi2 test = 24.956, P < 0.05). Compared with group A, the mRNA and protein expressions of HO-1 were elevated (F = 4.549, 22.926, P < 0.05) in group B and increased in group C-E, but there is no significant difference existed.</p><p><b>CONCLUSION</b>Blueberry may have preventive and protective effects on CCl4-induced hepatic fibrosis by reducing lipid peroxidation. However, these effects may not be related to the activation of HO-1 during long-term of CCl4.</p>


Subject(s)
Animals , Male , Rats , Blueberry Plants , Chemistry , Carbon Tetrachloride , Toxicity , Drugs, Chinese Herbal , Pharmacology , Heme Oxygenase (Decyclizing) , Blood , Liver Cirrhosis, Experimental , Blood , Malondialdehyde , Blood , Rats, Sprague-Dawley
7.
Chinese Journal of Endemiology ; (6): 604-607, 2010.
Article in Chinese | WPRIM | ID: wpr-642553

ABSTRACT

Objective To identify the differential serum proteins in patients with hepatic injury resulting from coal-burning type of arsenism. Methods Six serum samples were collected from patients with liver injury resulting from coal-burning type of arsenism and healthy subjects(control gruop) in endemic arsenism area. Twodimensional gel electrophoresis(2-DE) was performed to separate serum proteins, after silver staining, the differential expression of proteins were analyzed and then identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS). Results The 2-DE map of serum protein patterns of patients and normal control were established successfully. The results showed that there were an average of (824 ± 31 ) spots and (782 ± 42) spots on 2-DE matching of the patients and control groups and the matching rate was 94.9%(782/824). From these two groups 49 differential protein spots were identified, of which over 3 times the difference in the expression of 30 protein spots were singled out and MALDI-TOF-MS analysis was carried out. Ten proteins were identified. Upregulated expression was observed in alpha-2-macroglobulin, B-cell receptor-associated protein, keratin 1,apolipoprotein A-I, and down-regulated expression was observed in haptoglobin, α2-heremans-schimid-glycoprotein,mitogen-activated protein kinase 4, zinc finger protein 323, ZAP-70 and SP40 in the patient group. Conclusions The well-resolved and reproducible 2-DE serum patterns of patients are established and some differentially expressed proteins are characterized. Whether these proteins of differential expression are serum markers for liver injury resulting from coal-burning type of arsenism need to be further verified.

8.
Chinese Journal of Endemiology ; (6): 101-104, 2009.
Article in Chinese | WPRIM | ID: wpr-643278

ABSTRACT

Objective To summarize the prevention and control of endemic arsenism in Qianxinan state of Guizhou Province in 30 years,so as to provide scientific basis for prevention and control of arsenic poisoning. Methods According to the documentary information of endemic arsenism in Qianxinan state of Guizhou Province of 1976-2005.and the data from samples monitoring of published article,to discuss the general situation of prevention and control by time sequence.Results There were 3 large outbreaks in Xingren County in 1976.in Jiaole District in 1991,in Xingyi City and Anlong County in 1992-1994 with 877,1548 and 594 patients respcctively.The arsenic quantities of various areas of burning-coal were 3361,2130,352 and 2150 mg/kg,which were obviouslv above national standards(1 00 mg/kg).The arsenic quantities of samples were 1.34,1.34,0.71 and 0.39 mg/L in urine,and 53.50,53.50,58.60 and 12.60 mg/kg in hair.The national standards of the two samples were 0.17 mg/L and 0.28 mg/kg.The dead causes of 236 arsenism decedent were 45.3%(107/236)cancer,26.3% (62/236)hepatocirrhosis ascites,5.9%(14/230)lung-cardiopathy in 1992-2000.Other systemic damages were found.Conclusions Reducing the use of high-arsenic coal is crucial in prevention of endemic arsenism.Only the long-term mechanism is established and the strong prevention and control measures are formulated,the occurrence of endemic arsenism can be effectively controlled.

9.
Chinese Journal of Endemiology ; (6): 390-394, 2009.
Article in Chinese | WPRIM | ID: wpr-642310

ABSTRACT

Objective To study the expression and its significance of Platelet-derived growth factor(PDGF) and Collagen type Ⅰ (Col- Ⅰ) mRNA in hepatic tissue in mice exposed to arsenic in drinking water. Methods Fifty mice were divided into control group, sodium arsenite group(iAs3+ group, 300 mg/L) and sodium arsenate group (iAs5+ group, 300mg/L) at random. The mice were sacrificed after 10 months for liver function test (ALT, AST and GLB examined by automatic biochemical analyzer) and pathologic examinations. Masson staining and immunohistochemical SABC methods were used. Through the result of Masson staining fibrosis semi-quantitative analysis of the fibrosis number was carried out in the per unit area. Using immuonhistochemical SABC methods, the expression of PDGF and Col- Ⅰ was detected. Results ①Serum ALT was statistically significant among control group [(36.67±3.51) U/L], iAs3+ group[(61.46±13.85) U/L] and iAs5+ group[(43.31±4.21) U/L, F= 6.56, P < 0.05], especially between the control and iAs3+ group (P < 0.05) ;Serum AST had a statistical significance among control group [(135.00 ± 20.42)U/L], iA3+ group [(510.86 ± 59.01)U/L] and iAs5+ group[(258.93 ± 22.40) U/L, F= 83.36, P < 0.05] ;Serum GLB had no statistical significance among control group [(20.86 ± 0.61)g/L], iAs3+ group [(26.94± 3.73)g/L] and iAs5+ group[(24.59 ± 5.27)g/L, F = 2.80, P < 0.05]. ②Masson staining revealed notable liver cell necrosis, regeneration, infiltration of inflammatory cells in portal area. Masson staining Semi-quantitative results showed significantly increasing size of fibrosis in model group. Compared with control group(0.069 ± 0.013) and iAs5+ group(0.143 ± 0.122), fiber hyperplasia of iAs3+ group(0.192 ± 0.108) had statistical significance (F = 1.91, P < 0.05). ③Immunohistochemistry results showed higher expression of PDGF in the portal area, sepetal and preisinus cells in model group. The heavy expression of Col- Ⅰ distributed in the surrounding of blood vessels and bile duet and along portal wall extensting to the liver in model group. Semi-quantitative results showed that PDGF among control group(202.79 ± 7.46), iAs3+ group(174.38 ± 7.71) and iAs5+ group(177.64 ± 7.81) had a statistical significance(F= 102.91, P < 0.05);Col-Ⅰ had a statistical significance among control group(200.11 ± 7.46), iAs3+ group (176.47±10.20) and iAs5+ group (177.378 ±7.948, F = 78.51, P < 0.05). Conclusions Exposed in oral drinking arsenic, mice can develop hepatic injury and hepatic fibrosis. The PDGF and Col-Ⅰ play significant effects on the process of hepatic fibrosis caused by oral drinking arsenic exposed mice.

10.
Chinese Journal of Endemiology ; (6): 162-164, 2009.
Article in Chinese | WPRIM | ID: wpr-642236

ABSTRACT

Objective To observe expression of the keratin 10 (K10), c-myc and cox2 in buccal mucous membrane exfoliated cells, and changes of liver function of coal-burning arsenism patients, in order to explorate coal-burning arsenic poison circumference biology symbol. Methods The buccal mucous membrane exfoliated cells were collected from both arsenism patients and normal controls. Real-time PCR was employed to detect the changes of K10, c-myc, cox2 genes expression in these cells. Simultaneously, circumference venous blood was extracted and examinated liver function. Results K10 was found obviously overexpressed in arsenism patients(3.60±0.94) compared to control(1.82±0.68), the difference had statistics significance(t=2.15, P<0.05), c-mye and cox2 did not found obviously changed(c-myc: 3.50±2.77,3.39±2.07; cox2:5.90±1.40,4.73±1.91; t=1.26,1.65, P> 0.05). The serum ALT of patients(25.83±2A5) obviously increased than control(36.86±1735, t=2.55, P<0.05). Conclusions Expression of K10 gene in buccal mucous membrane cells may be regarded as sensitive molecular markers for skin pathologic changes in arsenic patients. The liver is a sensitive target organ of inorganic arsenic.

11.
Chinese Journal of Preventive Medicine ; (12): 29-32, 2007.
Article in Chinese | WPRIM | ID: wpr-290237

ABSTRACT

<p><b>OBJECTIVE</b>To probe into the situation and significance of p16 gene CPG island methylation in patients with arseniasis caused by coal-burning pollution.</p><p><b>METHODS</b>DNA was extracted using the Phenol-Chloroform method from leukocytes of 51 patients suffered from coal-burnt arsenism and 52 healthy volunteers. The quantity of the DNA was determined by UV spectrophotometry. Target DNA was denatured by NaOH, then the single strand DNA was modified by sodium bisulfite, converting all unmethylated (but not the methylated) cytosines to uracil. Subsequently a nested amplification with primers specific for methylated versus unmethylated DNA was performed, and PCR products were detected by gel electrophoresis.</p><p><b>RESULTS</b>Hypermethylation of the p16 CPG island was presented in 94.1% of the patients suffering from coal-burnt arsenism and in 73.1% of the healthy volunteers. There was statistical difference (P < 0.05) between them.</p><p><b>CONCLUSIONS</b>Methylation of p16 gene CPG island should have important pertinence in the metabolism of coal-burnt arsenism.</p>


Subject(s)
Humans , Arsenic Poisoning , Blood , Genetics , China , Coal , CpG Islands , DNA Methylation , Genes, p16
12.
Acta Physiologica Sinica ; (6): 190-196, 2007.
Article in Chinese | WPRIM | ID: wpr-258671

ABSTRACT

The present study was designed to observe the expressions of bone morphogenetic protein-7 (BMP-7) and inhibitory Smads in kidney of rats with diabetic nephropathy (DN), and explore the possible mechanism of DN. Male Wistar rats weighing 180-220 g were single injected with streptozocin (STZ, 55 mg/kg body weight) for 2, 4, 8 and 16 weeks to induce DN. Blood glucose, kidney weight/body weight and 24-hour urine protein in the control and DN rats were examined; the expressions of BMP-7, Smad6 and Smad7 were detected by using immunohistochemical techniques, Western blot and real-time PCR. The results showed that blood glucose and 24-hour urine protein in DN rats were higher than that in the control rats and kidney weight/body weight was also elevated in DN rats, especially in 16-week STZ-induced rats. The expressions of BMP-7 and Smad6 proteins in DN rats were elevated, while BMP-7 mRNA expression was increased 2 weeks after STZ injection and decreased 16 weeks after STZ injection. The expressions of Smad7 protein and mRNA were elevated in DN rats 2 weeks after STZ injection and decreased 16 weeks after STZ injection. In addition, the expressions of transforming growth factor-beta1 (TGF-beta1) and collagen type I (COL-I) mRNA were increased in DN rats. These results suggest in the early stage of DN, increase in BMP-7 and inhibitory Smad expression may play a role in the feedback regulation and restrain the development of DN.


Subject(s)
Animals , Male , Rats , Bone Morphogenetic Protein 7 , Genetics , Metabolism , Collagen Type I , Metabolism , Diabetes Mellitus, Experimental , Metabolism , Diabetic Nephropathies , Metabolism , Kidney , Metabolism , Pathology , RNA, Messenger , Genetics , Metabolism , Random Allocation , Rats, Wistar , Smad6 Protein , Genetics , Metabolism , Smad7 Protein , Genetics , Metabolism , Transforming Growth Factor beta1 , Metabolism
13.
Chinese Medical Sciences Journal ; (4): 243-249, 2007.
Article in English | WPRIM | ID: wpr-243520

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the role of transforming growth factor-beta1 (TGF-beta1)/Smad4 pathway in development of renal fibrosis in streptozotocin (STZ)-induced diabetic nephropathy (DN) rats and explore its possible mechanism.</p><p><b>METHODS</b>Male Wistar rats weighing 180-220 g were divided into 5 groups: group A (normal control), group B [diabetes mellitus (DM) 2 weeks], group C (DM 4 weeks), group D (DM 8 weeks), and group E (DM 16 weeks). Except for the normal control group, other groups were induced DM by single injection of STZ (55 mg/kg) respectively. Blood glucose level, serum creatinine, and 24-hour urine protein were examined. Expressions of TGF-beta1 and Smad4 protein and mRNA in kidney were detected using immunohistochemical technique, Western blot, and real-time PCR. mRNA expressions of stromelysin-1 (MMP-3), tissue inhibitor of metalloproteinase-1 (TIMP-1), and collagen In in kidney were also detected by real-time PCR.</p><p><b>RESULTS</b>The levels of blood glucose, serum creatinine, and 24-hour urine protein in rats of group B, C, D, and E were higher than those of the control group. With the progression of renal fibrosis, the expressions of TGF-beta1 and Smad4 protein and mRNA in kidney of diabetic rats elevated. In addition, the renal MMP-3 mRNA expression diminished in diabetic rats, while TIMP-1 and collagen III mRNA increased.</p><p><b>CONCLUSIONS</b>In STZ-induced diabetic rats, the TGF-beta1/Smad4 appears to play an important role in renal fibrosis of DN. The increased expression of TGF-beta1 and Smad4 might result in the transcriptional regulation of downstream target genes of TGF-beta1/Smad4 pathway, which contributes to the progression of renal fibrosis in diabetic rats.</p>


Subject(s)
Animals , Male , Rats , Base Sequence , DNA Primers , Diabetes Mellitus, Experimental , Metabolism , Down-Regulation , Extracellular Matrix , Metabolism , Kidney , Metabolism , Rats, Wistar , Signal Transduction , Smad4 Protein , Genetics , Metabolism , Transforming Growth Factor beta1 , Genetics , Metabolism
14.
Chinese Journal of Hepatology ; (12): 285-287, 2003.
Article in Chinese | WPRIM | ID: wpr-344420

ABSTRACT

<p><b>OBJECTIVE</b>To study the expression of connective tissue growth factor (CTGF) mRNA and transforming growth factor beta 1 (TGFbeta1) mRNA in immunity-induced liver fibrosis rats and the effect of HanDanGanLe on them.</p><p><b>METHODS</b>Male wistar rats were given intraperitoneal injection of porcine serum twice a week. At the beginning, the rats in HanDanGanLe-treatment group were feed with HanDanGanLe for precaution. The rats were killed after twelve weeks, then CTGF mRNA and TGFbeta1 mRNA were detected in liver samples with in situ hybridization, and the formation of liver fibrosis was observed with HE stain. The semi-quantitative</p><p><b>RESULTS</b>of the two genes expression were analysed along with the stages of hepatic fibrosis.</p><p><b>RESULTS</b>Typical liver fibrosis developed in the model group rats, and the positive stain of CTGF mRNA and TGFbeta1 mRNA increased, which was distributed in the areas where fibrosis occurred. There was obvious correlation between the expression strength of CTGF mRNA and TGFbeta1 mRNA (r = 0.799, P < 0.05). In the rats receiving HanDanGanLe, CTGF mRNA expression index decreased markedly (12.5+/-2.3 vs 28.8+/-1.4, t = 5.208, P < 0.01), so did TGFbeta1 mRNA expression index (25.4+/-3.2 vs 37.3+/-5.4, t = 5.655, P < 0.01). There was also significant correlation between the scores of CTGF mRNA expression and the stages of hepatic fibrosis (rs = 0.822, 0.808 in the model group and HanDanGanLe-treatment group, P < 0.05).</p><p><b>CONCLUSIONS</b>The expression of CTGF mRNA and TGFbeta1 mRNA is correlated closely with hepatic fibrosis degree. HanDanGanLe can effectively prevent the expression of CTGF and TGFbeta1. One of the mechanisms of the intervention may be its blocking the intracellular signalling pathways involved in liver fibrogenesis.</p>


Subject(s)
Animals , Male , Rats , Connective Tissue Growth Factor , Drugs, Chinese Herbal , Pharmacology , Immediate-Early Proteins , Genetics , Intercellular Signaling Peptides and Proteins , Genetics , Liver Cirrhosis, Experimental , Drug Therapy , Metabolism , RNA, Messenger , Genetics , Rats, Wistar , Signal Transduction , Transforming Growth Factor beta , Genetics , Transforming Growth Factor beta1
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