【Abstract】ObjectiveTo explore the effect of hepatocyte growth factor/scatter factor (HGF/SF) on apoptosis of colorectal cancer cells induced with curcumin. MethodsMTT assay was used to evaluate the cytotoxicity of curcumin to colorectal cancer cells. Flow cytometry was used to detect the antiapoptosis effect of HGF. ResultsFlow cytometry showed only 64 μg/ml curcumin could play the proliferationinhibiting role in Caco-2 cells leading to their apoptosis; at the same time, different concentrations of HGF could antagonize this inhibitory effect resulting in the decrease of apoptosis, but HGF worked without a concentration-dependent manner. The study on MAPK pathway showed that the protective effect of HGF on the apoptosis of Caco-2 cells was not influenced by inhibiting p42/p44 MAPK and p38 MAPK pathway. ConclusionHGF/SF antagonizes the apoptosis of Caco-2 cells induced with curcumin, but MAPK signaling pathway might not participate in this process.
ObjectiveTo study the protective effect of hepatocyte growth factor(HGF) on grafted mucous membrane of transplanted small bowel.MethodsTotal small bowel transplantation was made in inbred Wistar (RT1k) rats heterotopically,either total parenteral nutrition (control group,n=10) or hepatocyte growth factor supplemented TPN (HGF group,n=10) was given to the recipients from the 2nd day to the 10th day postoperatively. Morphological parameters of the transplanted intestinal mucosa, such as mucosal villous height, villous width, crypt depth, mucosal thickness and villous surface area were observed. Variation of ultrastructure of transplanted epithelial cells was observed. Composition of mucosal protein and DNA content were tested. ResultsComparison between HGF group and the control group were as follows. Mucosal villous height (298.79±22.31） μm vs （176.45±14.62） μm, P=0.001, villous width (107.46±12.34） μm vs （74.20±16.85） μm, P=0.004, crypt depth (104.56±11.17） μm vs （74.45±8.34） μm, P=0.000 5, mucosal thickness (409.53±35.83） μm vs （259.38±24.65） μm, P=0.003, and villous surface area (0.101±0.011) mm2 vs (0.041±0.005) mm2, P=0.001 were found significantly increased in HGF group compared with control group, there were no obvious difference decrease as compared to pretransplant parameters.Mucosal protein composition was higher in HGF group than that in control group （89.65±9.28) mg/g wet wt vs (53.73±11.45) mg/g wet wt, P=0.012, baseline (92.64±10.52) mg/g wet wt, nearly equal to baseline; DNA composition was also high in HGF group （0.89±0.09) mg/g wet wt vs (0.51±0.06) mg/g wet wt, P=0.008, baseline (0.91±0.09) mg/g wet wt. Nearly normal ultrastructure of the graft was maintained in HGF group, atrophied microvilli and broken mitochondrial crista were observed in control group.ConclusionHepatocyte growth factor can improve mucosal structure, preserve enterocyte ultrastructure of isograft after small bowel transplantation in rat.
ObjectiveTo introduce the relationship between the apoptosis hepatocyte and its genic mediation and the ischemia of portal vein. MethodsThe combination of related literatures and our research findings were made.ResultsPortal vein ischemia may induced hepatocyte apoptosis, p53 and bcl2 gene alternatively adjust hepatocyte apoptosis. Expression of p53 gene is enhanced in hepatic tissue when hepatocyte apoptosis is not obvious, but after 24-72 h of portal vein ischemia, when hepatocyte apoptosis is obvious, enhanced expression of p53 gene or reduced expression of bcl2 gene occur. There exists close relationship between portal vein ischemia and hepatocyte apoptosis. Conclusion Apoptosis hepatocyte is involved in organic atrophy after ischemia of portal vein, and p53 and bcl2 gene alternatively adjust hepatocyte apoptosis. At present, the mechanism of apoptosis of hepatocyte induced by ischemia of portal vein is not clear, which needs further study.
Objective To investigate the stable, high effective and low toxicity gene transfer vectors’ basics in research and clinical application.Methods Current status of hepatocytedirected gene transfer vectors were reviewed by history document and contemporary experimental advances analysis.Results Viral and nonviral vector systems could both transduce target genes to liver effectively with various transduction rates based on their corresponding biological characteristics.Conclusion Hepatocyte is the effective target for gene therapy of liverrelated genetic, neoplastic and infectious diseases, although the security needs to be further evaluated.
Objective To explore the effects and mechanism of autonomic nervous control on the proliferation of human hepatocytes. And to examine the cellular localization of some related receptors expression in human hepatocytes. MethodsNorepinephrine (NE), and its agonist, antagonist, acetylcholine (Ach), and its antagonist have been added to human hepatocyte line L02 and hepatoma cell line Bel7402. Modified MTT assay was employed to test the effects of them on the proliferation of the two cell lines at 4 h, 24 h, 48 h and 72 h. Immuocytochemical staining was used to examine the cellular localization of alpha1Badrenoceptor (α1BAR), β2AR and epidermal growth factor receptor (EGFR) expression in human hepatocyte line L02. ResultsNE potentiated the proliferation of human hepatocyte and hepatoma cell, which was enhanced significantly with dose increased. The proliferative rate of 4 h were higher than that of the other time points (P＜0.05). There were no significant differences between the group of NE combined with propanolol and the group of NE alone. Metaproterenol had no significant effect. Ach significantly inhibited the proliferation of human hepatocyte. Its effect was enhanced with dose increased. Atropine significantly attenuated the inhibitory effect of Ach at 24 h and 48 h (P＜0.05). Scoline alone inhibited hepatocyte proliferation at 24 h, 48 h and 72 h (P＜0.05, P＜0.01). In immunocytochemical staining, there were positive responses to α1BAR, β2AR and EGFR in all cultures. It was observed that the responses to α1BAR, β2AR and EGFR were mainly both cytoplasmic and cell membrane localized. Conclusion NE, the sympathetic neurotransmitter, acts via α1BAR potentiate the proliferation of human hepatocyte and hepatoma cell in the presence of serum. Ach, the vagus neurotransmitter acting via mAchR and nAchR inhibits hepatocyte proliferation.
【Abstract】Objective To search for the drug that promots the hepatocyte regeneration after partial hepatectomy. Methods Quantitative morphometry technique, 3H-TdR in vivo test and arterial ketone body ratio (AKBR) were employed to evaluate the energy metabolism, DNA synthesis, and liver cell nuclear mitosis and hepatocyte regeneration of the residual liver tissue in rats after partial lobectomy when treated with human growth hormone (hGH) and placebo. Results After partial hepatectomy in the experimental group, liver cell nuclear mitosis，nuclear density, new liver cell numbers, AKBR and 3H-TdR in vivo test were much higher than those in the control group (P＜0.01 or P＜0.05). Conclusion hGH promotes the hepatocyte regeneration of the residual liver.
【Abstract】Objective To investigate the injury of calcium on the liver. Methods By using collagenase-containing solution to perfuse the rat livers, the rat liver suspension with different viability was prepared, preserved hypothermically, and the cytosolic calcium comcentration was detected with Fura2/AM. Results ① The concentration of cytosolic calcium after 2-hour storage: Experiment group 1(viability 5%) (1055.0±30.79) nmol/L, experiment group 2(viability 10%) (853.0±20.42) nmol/L, experiment group 3(viability 30%) (616.0±13.20) nmol/L, experiment group 4(viability 50%) (562.0 ±26.06) nmol/L, experiment group 5(viability 70%-80%) (318.0±13.01) nmol/L, experiment group 6(viability 90%) (114.6±6.11) nmol/L. ②The concentration of cytosolic calcium after 24hour storage: Experiment group A(viability 10%) (1704.0±70.11) nmol/L, experiment group B(viability 50%) (1125.0±23.22) nmol/L. The results showed that the lower was the viability, the higher was the cytosolic calcium concentration.With the same viability the cytosolic calcium concentration elevated more than two times higher than the original concentration with the time lengthened. Conclusion Calcium overload is one of the main factors which attribute to the ischemiareperfusion injury of the hepatocytes.
In order to observe the effect of hepatocyte growth promoting factor (pHGF) on liver regeneration of rat with cirrhosis after hepatectomy, IBAS Ⅱ auto image analysis technology was used to measure the variety of DNA ploid rate of hepatocytes and OPTDM of enzymes by liver histochemistry after hepatectomy; serum levels of the glutamicpyruvic transaminase (SGPT) and indocyanine green retention rate in 15 minute (ICG15) were tested to measure the function of the remanent liver. The results revealed that tetraploid hepatocytes lowered greatly and diploid, quintploid and ＞quintploid hepatocytes increased apparently in group A. OPTDM of enzymes by liver histochemistry showed no significant difference at the first day after operation in each group (Ｐ＞0.05); SDH and LDH of group A were significantly higher than those of group B and AkP, AcP were significantly lower at the second or fifth day after hepatectomy. Serum tests showed that SGPT, ICG15 of group A decreased apparently at the fifth day after operation. The results demonstrate that pHGF not only stimulates the regeneration of the remanent liver but also accelerates the functional mature of the regenerative hepatocytes and the functional recovery of the remanent liver after resection of cirrhotic liver of rats.
Objective To investigate the possible association between serum level of hepatocyte growth factor( HGF) and obstructive sleep apnea hypopnea syndrome( OSAHS) with hypertension.Methods 58 cases of OSAHS without hypertension, 61 cases of OSAHS with hypertension, and 50 normal controls were enrolled. Serum level of HGF was measured by enzyme-linked immunosorbent assay( ELISA) , and the relationships between the serum HGF level and blood pressure( BP) , apnea hypopnea index( AHI) , lowest SaO2 ( LSaO2 ) were analyzed by linear correlation analysis. Results The serum HGF level ( pg/mL) was 761. 46 ±60. 18, 970. 87 ±60. 94, and 487. 34 ±45. 52 in the OSAHS patients without hypertention, OSAHS patients with hypertention, and normal subjects, respectively. Which was significantly higher in the OSAHSpatients than the normal subjects, and highest in the OSAHS patients with hypertension( P lt; 0. 05) . The serum HGF level was positively related to AHI( r = 0. 452, P lt;0. 05) and negatively related to LSaO2 ( r =- 0. 328, P lt;0. 05) in the OSAHS patients without hypertention, positively related to AHI, SBP, DBP( r =0. 670, P lt;0. 01; r =0. 535, P lt;0. 05; r =0. 424, P lt;0. 05) and negatively related to LSaO2 ( r = - 0. 572,P lt;0. 01) in the OSAHS patients with hypertension. Conclusions SerumHGF level increases significantly in patients with OSAHS especialy in OSAHS patients with hypertension, and positively correlates with the severity of OSAHS and hypertension.
【Abstract】 Objective To explore the new therapy for pulmonary fibrosis by observing the effects of insulin-like growth factor 1 ( IGF-1) treated mesenchymal stemcells ( MSCs) in rats with bleomycin-induced pulmonary fibrosis. Methods Bone marrowmesenchymal stemcells ( BMSCs) were harvested from6-week old male SD rats and cultured in vitro for the experiment. 48 SD rats were randomly divided into 4 groups, ie.a negative control group ( N) , a positive control group/bleomycin group ( B) , a MSCs grafting group ( M) ,and an IGF-1 treated MSCs grafting group ( I) . The rats in group B, M and I were intratracheally injected with bleomycin ( 1 mL,5 mg/kg) to induce pulmonary fibrosis. Group N were given saline as control. Group M/ I were injected the suspension of the CM-Dil labled-MSCs ( with no treatment/pre-incubated with IGF-1 for 48 hours) ( 0. 5mL,2 ×106 ) via the tail vein 2 days after injected bleomycin, and group B were injected with saline ( 0. 5 mL) simultaneously. The rats were sacrificed at 7,14,28 days after modeling. The histological changes of lung tissue were studied by HE and Masson’s trichrome staining. Hydroxyproline level in lung tissue was measured by digestion method. Frozen sections were made to observe the distribution of BMSCs in lung tissue, and the mRNA expression of hepatocyte growth factor ( HGF) was assayed by RTPCR.Results It was found that the red fluorescence of BMSCs existed in group M and I under the microscope and the integrated of optical density ( IOD) of group I was higher than that of group M at any time point. But the fluorescence was attenuated both in group M and group I until day 28. In the earlier period, the alveolitis in group B was more severe than that in the two cells-grafting groups in which group I was obviously milder. But there was no significant difference among group I, M and group N on day 28.Pulmonary fibrosis in group B, Mand I was significantly more severe than that in group N on day 14, but itwas milder in group M and I than that in group B on day 28. Otherwise, no difference existed between the two cells-grafting groups all the time. The content of hydroxyproline in group B was significantly higher than that in the other three groups all through the experiment, while there was on significant difference betweengroup I and group N fromthe beginning to the end. The value of group M was higher than those of group I and group N in the earlier period but decreased to the level of negative control group on day 28. Content of HGF mRNA in group Nand group I was maintained at a low level during the whole experiment process. The expression of HGF mRNA in group I was comparable to group M on day 7 and exceeded on day 14, the difference of which was more remarkable on day 28. Conclusions IGF-1 can enhance the migratory capacity of MSCs which may be a more effective treatment of lung disease. The mechanismmight be relatedto the increasing expression of HGF in MSCs.