Serum GP73 were increased in patients with fatty liver disease / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore the levels of serum GP73 in patients with fatty liver disease.</p><p><b>METHODS</b>The sera GP73 were determined by ELISA in 178 patients with fatty liver disease and 100 healthy controls.</p><p><b>RESULTS</b>Serum GP73 levels were significantly increased in patients with various fatty liver diseases(70.62 +/- 60.60 ng/ml), compared with those of control population (35.61 +/- 12.22 ng/ml). In patients with alcoholic fatty liver disease, acute liver injury, chronic hepatitis B, and non-alcoholic fatty liver disease, their serum GP73 concentration were 81.86 +/- 47.82 ng/ml, 82.77 +/- 77.73 ng/ml, 63.84 +/- 50.62 ng/ml, and 65.75 +/- 62.20 ng/ml, respectively. But no significant difference was found between these groups (P > 0.05). In 68 patients with F > or = 1.0 (71.46 +/- 66.48 ng/ml), 75 patients with F> or = 2.0 (69.58 +/- 62.31 ng/ml), and 34 patients with F3-F4 (71.65 +/- 43.89 ng/ml), there were also no marked differences was observed between these fatty groups (F = 0.02, P = 0.98).</p><p><b>CONCLUSION</b>Serum GP73 levels were increased in patients with different liver diseases, but its concentrations were seems not related with degree of fatty injury.</p>

Establishment of a detection method for hepatitis B virus large surface protein (HBV-lP) / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish enzyme-linked immunosorbent assay (ELISA) for detection of hepatitis B virus large surface protein(HBV-LP) in serum.</p><p><b>METHODS</b>A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of HBV-LP as the catalytic enzyme. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as sensitivity, specificity, stability and so on.</p><p><b>RESULTS</b>The detection limit was 5 ng/ml. Interassay and intra-assay RSD were both less than 10%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.98 and RSD lower than 10%.</p><p><b>CONCLUSION</b>Established ELISA for determination of serum HBV-LP has high sensitivity and repeatability. Enzyme-linked immunosorbent assay;</p>

Microplate chemiluminescence enzyme immunoassay for the quantitative analysis of tissue inhibitor of metalloproteinases I / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish microplate chemiluminescence enzyme immunoassay (CLEIA) for quantitative analysis of tissue inhibitor of metalloproteinases I (TIMP I) in human serum.</p><p><b>METHODS</b>A sandwich reaction was preformed with horseradish peroxidase(HRP) labeled monoclonal antibody of TIMP I as the catalytic enzyme and the H2O2-luminol as the luminescence reagent. Several physical and chemical parameters were studied and optimized such as immunoreaction conditions, the dilution ratio of TIMP I-HRP, luminescence reaction time and so on. In order to evaluate the method, recovery test, heat stabilization test and comparison test were carried out.</p><p><b>RESULTS</b>The linear range was 0. 2-12 ng/ml with r = 0.996. The detection limit was 0.12 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 100.6%, 96.5% and 106.5%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0. 998 and RSD lower than 6%. The detected results with CLEIA closely corresponded to those with imported ELISA in 60 patients sera with liver fibrosis.</p><p><b>CONCLUSION</b>Established CLEIA for quantity determination of serum TIMP I has high accuracy, sensitivity and repeatability.</p>

Cloning and expressing of golgi protein73 gene fragment and preparation of monoclonal antibodies against the recombinant protein / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To clone and express human Golgi glycoprotein73 protein, and prepare the monoclonal antibody (mAb) against the protein.</p><p><b>METHODS</b>GP73 gene was amplified from HepG2 cells by RT-PCR, then ligated with pQE31 to form recombinant plasmid pQE-GP73 and transformed into E. coli BL21. The protein induced by IPTG was purified by 6 x His-tag and used to immunize the BALB/c mice. The specific monoclonal antibodies (mAbs) were prepared by the cell fusion technique. Western Blot was used to detect specificity of mAbs.</p><p><b>RESULTS</b>The prokaryotic plasmid expressing the recombinant protein was constructed, and the GP73 recombinant protein was expressed and purified. Five hybridoma cell lines that secreted anti-GP73 mAbs were obtained. 2 of 5 mAbs were the IgG1 subtype. Western Blot indicated the mAbs showed specific combination with GP73 protein.</p><p><b>CONCLUSION</b>The GP73 recombinant protein is highly purified and has strong antigenicity. The anti-GP73 mAbs were prepared successfully.</p>

Establishment of a quantitative detection method for Golgi protein73 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish enzyme-linked immunosorbent assay (ELISA) for quantitative detection of Golgi protein73 (GP73) in serum.</p><p><b>METHODS</b>A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of GP73 as the catalytic enzyme. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as linear range, sensitivity, specificity, stability and so on.</p><p><b>RESULTS</b>The linear range was 25-500 ng/ml. The detection limit was 18.5 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 95.3%, 92.6% and 103.7%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.98 and RSD lower than 10%.</p><p><b>CONCLUSION</b>Established ELISA for quantity determination of serum GP73 has high accuracy, sensitivity and repeatability.</p>

Quantitative detection of serum procollagen III N-terminal peptide chemiluminescence enzyme immunoassay / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish chemiluminescence enzyme immunoassay (CLEIA) for quantitative detection of procollagen III N-terminal peptide (P III NP) in serum.</p><p><b>METHODS</b>A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of P III NP as the catalytic enzyme and the luminol as the luminescence reagent. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as linear range, sensitivity, specificity, stability and so on. The CLEIA was compared with imported ELISA kits, by detecting clinical serum.</p><p><b>RESULTS</b>The linear range was 0.8-85 ng/ml. The detection limit was 0.5 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 96.2%, 91.2% and 101.1%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.99 and RSD lower than 6%. The detected results of clinical sera with CLEIA closely corresponded to those with imported ELISA.</p><p><b>CONCLUSION</b>Established CLEIA for quantity determination of serum P III NP has high accuracy, sensitivity and repeatability.</p>

Role of C6ORF120, an N-glycosylated protein, is implicated in apoptosis of CD4⁺ T lymphocytes / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Although CD4(+) T cell apoptosis and CD8(+) T cell responses have been extensively studied during HIV infection, how apoptosis signals being initiated in CD4(+) T cells still need to be elucidated. The present study was designed to characterize the function-unknown gene, C6orf120, and elucidates its primary role in tunicamycin-induced CD4(+) T apoptosis.</p><p><b>METHODS</b>The C6orf120 coding sequence was amplified from peripheral blood mononuclear cells (PBMCs) total RNA of AIDS patients. The DNA fragment was inserted into the pET-32a expression system, transformed into Escherichia coli, and preparation of C6ORF120 recombinant protein. The magnetic cell separation technology was used to prepare primary CD4(+) T cells and CD8(+) T cells. The primary T cells were cultured at 1 × 10(6) cells/ml, treated with 0, 0.1, 1, 10, 100, and 200 ng/ml of C6orf120 recombinant protein for 48 hours, then harvested for cell cycle and apoptosis analysis. Tunicamycin (0.5 µmol/L) was used to induce endoplasmic reticulum stress in Jurkat cells. The biomarker 78 KDa glucose-regulated protein (GRp78) and growth arrest and DNA damage (GADD) were used to evaluate endoplasmic reticulum stress of Jurkat cells.</p><p><b>RESULTS</b>We prepared C6ORF120 recombinant protein and its polyclonal antibody. Immunohistochemical analysis showed that C6orf120 mainly expressed in hepatocytes and cells in germinal center of lymph node. At concentration of 0.1, 1, 10, 100, and 200 ng/ml, C6orf120 recombinant protein could induce apoptosis of Jurkat cells and primary CD4(+) T cells, and promoting G2 phase of its cell cycle. Western blotting analysis showed that C6ORF120 recombinant protein increased the expression of GRp78 and GADD in Jurkat cells in vitro.</p><p><b>CONCLUSION</b>Our results suggested that C6ORF120 could induce apoptosis of CD4(+) T cells, at least in part, mediated with endoplasmic reticulum stress.</p>

Relationship of CD4+ CD25hi regulatory T (Treg) cells to disease progression in HIV-infected patients / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Analyzing the relationships between peripheral blood CD4+ CD25hi regulatory T (Treg) cells and peripheral blood immune status or plasma HIV-lviral load in HIV-infected individuals,so as to determine whether Treg were related to the progression of HIV-infected disease.</p><p><b>METHODS</b>116 HIV-infected patients in different stages and 21 healthy control individuals were included in this study. The CD4+ and CD8+ T cell counts were determined by a standard 4-color flow cytometry technique. The Treg cells were examined with 3-color immune staining flow cytometry. The plasma HIV-1 viral load was detected by real time PCR.</p><p><b>RESULTS</b>The frequencies of Treg cells decreased in HIV-infected individuals with high CD4+ T cell counts( > 300/microl) compared with normal controls. With the progression of disease the frequencies of Treg cells were raised gradually, until were increased in HIV-infected individuals with low levels of CD4+ T cell counts ( < 100/microl). In addition, the frequencies of Treg cells were inversely related to CD4+ T cell counts and CD4+ /CD8+ ratio, data showed a statistically significant (respectively, r = -0.564, P < 0.001; r = -0.377, P < 0.001). Furthermore, the proportions of Treg cells were closely related to plasma HIV-1 RNA viral load (r = 0.514, P < 0.001).</p><p><b>CONCLUSION</b>CD4 CD25hi Treg cells should be a kind of important cells participating the immunopathogenesis of AIDS. It may play different roles in different stages of HIV-infected disease. The exact mechanism of Treg cells in the progression of the HIV-infected disease needs to be investigate further.</p>

Different plasma levels of interleukins and chemokines: comparison between children and adults with AIDS in China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The immunological differences between children and adults with AIDS in China are not well documented. Th1/Th2 cytokines and chemokines are two types of immune factors intimately involved in disease progression of HIV-1 infection. This study aimed to identify changes in plasma levels of Th1/Th2 cytokines inerleukin (IL)-18, IL-16, IL-10 and chemokines regulated on activation, normal T cell expressed and secreted (RANTES), stromal cell-derived factor-1 (SDF-1) and monocyte chemoattractant protein-1 (MCP-1) in HIV-1-infected children and adults in China.</p><p><b>METHODS</b>Seventy-five children with AIDS and 35 adult AIDS patients were recruited and clinical data were collected. CD4(+) T lymphocyte counts were measured by flow cytometery and plasma HIV RNA levels were measured by quantitative RT-PCR. Plasma levels of IL-18, IL-10, IL-16, RANTES, MCP-1, SDF-1alpha and SDF-1beta were quantified by enzyme-linked immunosorbent assay. The levels of beta2-microglobulin (beta2-MG) and soluble Fas (sFas) were measured to validate the level of humoral and cellular immune activation.</p><p><b>RESULTS</b>The mean levels of all cytokines in pediatric and adult AIDS patients were significantly higher than in their healthy controls (P < 0.01). The mean levels of these cytokines were higher in pediatric patients than in adult patients (P < 0.05, except for SDF-1alpha and beta2-MG). Some of the cytokine levels in patients younger than 6 years old was higher than in older children and adults with AIDS (IL-10, IL-18, SDF-1alpha, MCP, RANTES and sFas, P < 0.05). Levels of IL-18, IL-10, RANTES and beta2-MG of pediatric patients increased as the levels of viral load increased (P < 0.05).</p><p><b>CONCLUSIONS</b>Abnormal immune activation can be measured in Chinese pediatric and adult patients with AIDS, and is higher in children than in adult patients. The cytokines levels coincide with disease progression of AIDS, but have no direct relationship with total CD4(+) T cell count.</p>

Expression, purification and polyclonal antibody preparation for a novel gene BC097361 / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To express and purify of the BC097361 recombinant protein, and to prepare the BC097361 specific rabbit polyclonal antibody.</p><p><b>METHODS</b>BC097361 cDNA was ligated into the prokaryotic expressive vector pET-32a (+), and the resulting plasmid was transformed into E.coli BL21 (DE3). The protein expression was induced with IPTG and the protein was analyzed with SDS-PAGE and western blotting. The expressed product was purified using Ni+ affinity column chromatography.Then the purified pET-32a (+) -BC097361 fusion protein was used to immunize New Zealand rabbits to gain polyclonal antibody. The specificity and potency of polyclonal antibody were evaluated by Western blot and ELISA.</p><p><b>RESULTS</b>The BC097361 fusion protein was highly expressed.The protein was mainly in the inclusion body. ELISA indicated the titer of polyclonal antibody more than 1:320000. The high specificity was confirmed with Western blot.</p><p><b>CONCLUSIONS</b>The recombinant BC097361 fusion protein and the BC097361 specific polyclonal antibody will be valuable tools for the investigation on the biological function of BC097361.</p>

Impact of highly active antiretroviral therapy on plasma MCP-1 and MSP in AIDS patients / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To study the effect of highly active antiretroviral therapy (HAART) on plasma levels of MSP and MCP-1 in AIDS patients.</p><p><b>METHODS</b>Forty Chinese AIDS patients were treated with HAART for 3 months and 84 German AIDS patients with HAART for 3 to 6 years. The pre-treatment and post-treatment plasma levels of MSP and MCP-1 were measured by enzyme-linked immunosorbent assay (ELISA), and their correlations with CD4+ cell counts and viral loads were analyzed.</p><p><b>RESULT</b>The mean levels of MCP-1 were significantly higher and MSP were significantly lower in HIV-infected patients compared with the HIV-negative controls (P <0.01). After HAART for three months, there were no significant changes in the levels of these cytokines. But after long-term HAART (for 3 to 6 y), the level of MCP-1 was increased and that of MSP decreased significantly (P<0.01). There was a negative correlation between MSP and MCP-1 levels, and the same for MSP level and CD4+ cell counts; while there was a positive correlation between MCP-1 levels and CD4+ cell counts.</p><p><b>CONCLUSION</b>The changed plasma levels of MSP and MCP-1 are associated with HIV-1 infection and HAART may reverse the levels of these two cytokines.</p>

The association of TGF beta1 and AT1R gene polymorphisms with hereditary susceptibility and clinical phenotype of HBV-induced liver cirrhosis / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the association of TGF beta1 and AT1R gene polymorphisms with hereditary susceptibility and clinical phenotype of HBV-induced liver cirrhosis.</p><p><b>METHODS</b>Peripheral blood samples were collected from 102 patients with HBV-induced liver cirrhosis and 106 healthy blood donors. The polymorphisms of the promoter site -509C/T of TGF beta1 and 1166A/C of AT1R gene were determined by PCR-RFLP.</p><p><b>RESULTS</b>The frequency of the homozygote CC of -509C/T of TGF beta1 gene in the group of liver cirrhosis was higher than that the control group (P<0.05); and the frequency rate of homozygote CC was higher in group C than in group A and group B of liver cirrhosis (P<0.05), but there was no significant difference in allele frequency among these group (P>0.05). There was no significant difference in genotypes and allele frequency of AT1R gene 1166A/C between the liver cirrhosis group and the control group (P>0.05).</p><p><b>CONCLUSION</b>The polymorphism of the promoter site -509C/T of TGF beta1 gene is associated with hereditary susceptibility to liver cirrhosis and severity of HBV-induced liver cirrhosis; the polymorphism of AT1R gene 1166A/C is not associated with hereditary susceptibility to HBV-induced liver cirrhosis.</p>

Comparison of immunological profiles between pediatric and adult patients with AIDS in China / 中国医学科学院学报

<p><b>OBJECTIVE</b>To compare the immunological profiles of pediatric and adult patients with AIDS in China.</p><p><b>METHODS</b>Totally 103 pediatric AIDS patients, 38 adult patients, 88 healthy children, and 72 healthy adults were enrolled. CD4 + T lymphocyte counts were determined by four-color flow cytometer and HIV-RNA levels were measured in EDTA plasma by quantitative reverse-transcription polymerase chain reaction (RT-PCR). Plasma levels of interleukin (IL)-10, IL-16, IL-18, regulated upon activation, normal T-cell expressed and secreted (RANTES), monocyte chemoattractant protein 1 (MCP-1), stromal cell-derived factor-(SDF-1) alpha, SDF-1 beta, and macrophage stimulate protein (MSP) were quantified by enzyme-linked immunosorbent assay (ELISA). The levels of beta 2-microglobulin (beta 2-MG) and soluble Fas (sFas) were measured to indicate the activation of immune system.</p><p><b>RESULTS</b>The mean CD4 + T cell count in pediatric patients with AIDS was significantly lower than in healthy children (P < 0.01), as between the adult AIDS patients and healthy adults (P < 0.01). The mean levels of these cytokines in pediatric patients were significantly higher than in healthy children (P < 0.01). The level of MSP in adult patients was significantly lower than in healthy adults and other cytokines were significantly higher (P < 0.01). The mean levels of these cytokines, except SDF1 alpha and beta 2-MG, were significantly higher in pediatric patients than in adult patients (P < 0.01).</p><p><b>CONCLUSION</b>Abnormal immune activation is induced in both pediatric and adult patients with HIV-1 infection. The level of immune activation is higher in pediatric patients than in adult patients.</p>

Effects of traditional Chinese medicine on CD4 + T cell counts and HIV viral loads during structured treatment interruption in highly active antiretroviral therapy / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the impacts of traditional Chinese medicine (TCM) on CD4 + T cell counts and human immunodeficiency virus (HIV) viral loads during the course of structured treatment interruption (STI) in highly active antiretroviral therapy (HAART).</p><p><b>METHODS</b>Nineteen HIV/ADIS patients were treated for 14 months as follows: initiated with zidovudine/lamivudine + efavirdine for 6 months, then discontinued the therapy and treated with TCM instead for 2 months. HAART was then reinitiated for another 3 months, and then discontinued and replaced with TCM for another 3 months. The changes of CD4 + T cell counts and HIV viral loads were measured.</p><p><b>RESULTS</b>During the first STI of HAART, 43.8% of patients had no viral rebounds one month later, and 62.6% had stable or increased immune functions; 18.8% had no viral rebounds two months later, and 43.8% had stable or increased immune functions. Changes of viral loads were not significantly different between these two months (P = 0.097), while CD4 + T cell counts significantly decreased two months later compared with one month later (P = 0.043). During the second STI of HAART, 33.3% of patients had no viral rebounds one month later, and 64.3% had stable or increased immune functions; 13.3% had no viral rebounds 3 months later and 46.6% had stable or increased immune functions. Changes of viral loads had significant difference (P = 0. 017), while CD4 + T cell counts at month 12 elevated significantly compared with the baseline (P = 0.014).</p><p><b>CONCLUSIONS</b>TCM can suppress the viral rebounds during STI-HAART, maintain immune functions. However, this effect may decrease along with the prolongation of STI-HAART.</p>

The effects of angiotensin II on gene expression of hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>Our previous investigation demonstrated that angiotensin II could induce proliferation and differentiation of hepatic stellate cells, and also could up-regulate its extracellular matrix synthesis. The objective of this study was to determine the effects of 10(-5) mol/L angiotensin II on gene expression of hepatic stellate cells.</p><p><b>METHODS</b>After incubation with 10(-5) mol/L angiotensin II for 48 hours, the cultured hepatic stellate cells were collected. The mRNA and total protein were obtained from cell lysate and then suppression subtractive hybridization (SSH), 2D-gel electrophoresis and MALDI-TOF mass spectrometry were used to identify these cDNAs and proteins.</p><p><b>RESULTS</b>A total of 36 clones from the subtracted cDNA library were sequenced and compared to sequences in the GenBank using BLAST. Of the 36 differentially expressed gene fragments from the subtracted library, 13 differentially expressed gene fragments showed significant homology to other known proteins, such as ribosomal protein, beta-actin FE-3, leucyl-tRNA synthetase, CD147, pyruvate kinase, peroxiredoxin 1, and BAT3, while 2 other gene fragments encoding protein BC097361 and BC057380 and their functions were not disclosed. About 1110 and 1008 protein spots were detected by employing the ImageMaster 2D Platinum 4.9 proteome image analysis system in angiotensin-treated hepatic stellate cells and control cells separately. Among these spots 108 proteins were up-regulated while the other 153 proteins were down-regulated. Several up-regulated proteins were chosen to be excised and in-gel digest MALDI-TOF-MS and Database analysis showed that among the high expression proteins, there were prohibitin, RBL-NDP kinase 1.8x10(4) subunit, electron transfer flavoprotein alpha-subunit, guanine nucleotide binding protein, alpha 15, and heat shock 7.0x10(4) protein 5.</p><p><b>CONCLUSION</b>Our results suggest that the up-regulation of hepatic stellate cell mRNA influences proliferation, differentiation and apoptosis of those cells. The proteins of signal transduction, metabolizing regulation, apoptosis suppression, and fibrogenesis regulation of hepatic stellate cells were up-regulated.</p>

Relationship between B virus hepatitis genotypes and therapeutic efficacy in early treatment for chronic hepatitis B by using lamivudine / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between hepatitis B virus (HBV) genotype and therapeutic efficacy during the early phase of lamivudine treatment.</p><p><b>METHODS</b>Totally 595 patients with chronic hepatitis B were treated with lamivudine 100 mg/day for 12 months. HBV genotypes, contents of HBV DNA, HBeAg/anti-HBe and YMDD mutation after lamivudine treatment for 12 months were determined. The data were analyzed with SPSS software.</p><p><b>RESULTS</b>In 595 patients, 8 (1.4%) were genotype A; 53 (8.9%) genotype B; 360 (60.5%) genotype C; 112 (18.8%) were coinfection of genotype B and C; 14 (2.4%) of A and C; 15 (2.5%) A and B; 6 (1.0%) of A, B, and C, and remaining 27 (4.5%) were unspecified. Patients were treated with lamivudine 100 mg/day for 12 months. Genotype B with HBV DNA levels turned to be negative (HBV DNA < 0.1 ng/L) was 87.2%, genotype C was 89.51%, coinfection of genotype B and C was 93.04% (P > 0.05). HBeAg seroconversion of genotype B was 11.65%, of genotype C was 20.64%, and of coinfection of genotype B and C was 18.57% (P > 0.05). All 69 strains of YMDD mutation were detected after lamivudine treatment for 12 months, in which genotype B was in 16.98%, genotype C in 15.38%, and coinfection of genotype B and C was in 13.86% (P > 0.05).</p><p><b>CONCLUSION</b>There was no difference in HBV genotypes and the rate of development of YMDD mutations, HBeAg seroconversion, descending of HBV DNA level in Chinese patients with chronic hepatitis B.</p>

Therapeutic effects of octreotide on hepatofibrosis-induced with carbon tetrachloride in rats / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the therapeutic effects and mechanism of octreotide on experimental hepatic fibrosis in rats.</p><p><b>METHODS</b>Hepatofibrotic rats models were established with carbon tetrachloride. All the experimental rats were divided into four groups: normal control group, pre-and post-treatment model group, and octreotide-treated group in which the rats were injected subcutaneously with octreotide at the dose of 50ng/100g, twice daily, for thirty days. Serum levels of hyaluronic acid (HA), laminin (LN) and pro-collagen type III peptide (PCIII) were detected by radioimmunoassay. Hepatic fibrosis scoring grade was assessed through Van-Gieson staining and observed under light microscope. Protein expression levels of alpha-smooth muscle actin (alpha-SMA) and transforming growth factor beta1 (TGFbeta1) were determined with immunohistochemical staining method. Messenger RNA (mRNA) levels of collagen type I and PCIII were detected by reverse transcription polymerase chain reaction.</p><p><b>RESULTS</b>Serum levels of HA (ng/L), LN (microg/L) and PCIII (ng/L) in pre- and post-treatment model groups were higher than those in normal control group (121.8+/-9.5 and 110.3+/-13.4 vs. 33.1+/-3.7, 85.7+/-12.1 and 78.2+/-7.9 vs. 37.1+/-6.3, 35.9+/-3.5 and 33.7+/-2.6 vs. 15.6+/-2.8, respectively, t > or = 9.41, P<0.05), and there was no significant difference between the two model groups. Concentrations of HA (55.8ng/L+/-7.2ng/L), LN (43.1microg/L+/-3.4microg/L) and PCIII (27.8ng/L+/-3.4ng/L) decreased significantly in octreotide-treated group, compared with those in model groups (t >or=2.76, P<0.05). With histological analysis, fibrotic scoring grade in octreotide-treated group was obviously ameliorated, compared with that in model groups (chi2 > or = 3.97, P<0.05). Imaging analysis revealed that alpha-SMA and TGFbeta1 immunohistological staining areas were markedly shrinked in octreotide-treated group (t > or = 2.47, P < 0.05). In two model groups, PCIII and type I mRNA levels significantly up-regulated as compared with those in normal group (t > or = 9.27, P<0.001), and they were inhibited by octreotide markedly (t > or = 2.47, P<0.05).</p><p><b>CONCLUSIONS</b>Octreotide can inhibit hepatic stellate cells transforming into myofibroblasts, down-regulate TGFbeta1, collagen type I and PCIII transcriptions, so that it has therapeutic effects on experimental hepatic fibrosis.</p>