目的 综合评价中国乙型肝炎肝硬化人群并发原发性肝癌的相关危险因素.方法 计算机检索英文数据库PubMed、EBSCO、Highwire、Web of Science及中文数据库中国知网(CNKI)、万方数据库、维普期刊资源整合服务平台(VIP)中2005至2018年与乙型肝炎肝硬化并发原发性肝癌相关因素有关的研究或分析,同时手工检索相关资料.严格按照纳入和排除标准对所获文献进行筛选、提取资料和质量评价,完成后应用Stata 14.0进行Meta分析.结果 最终纳入文献共12篇,其中中文文献11篇,英文文献1篇;纳入人群总数2313例,病例组1171例,对照组1142例.M eta分析显示,乙型肝炎肝硬化患者的性别、饮酒史、吸烟史、肝癌家族史、糖尿病病史、不良饮食习惯、未经抗病毒治疗以及HBV DNA水平、HBsAg阳性、HBeAg阳性、抗-HBe阳性与乙型肝炎肝硬化并发原发性肝癌有关,各因素分析合并OR值及其各自95％C I值分别为1.30[1.03～1.65]、2.82[1.86～4.29]、2.98[1.62～5.48]、4.36[2.54～7.49]、2.79[2.04～3.82]、2.39[1.58～3.62]、3.74[2.86～4.87]、3.93[2.65～5.83]、5.52[3.61～8.43]、4.18[3.08～5.66]、2.59[1.91～3.53].结论 性别、饮酒史、吸烟史、肝癌家族史、糖尿病病史、不良饮食习惯、未经抗病毒治疗以及HBV DNA水平、HBsAg阳性、HBeAg阳性、抗-HBe阳性是导致乙型肝炎肝硬化患者并发肝癌的危险因素.

OBJECTIVETo investigate the effects of blueberry on the apoptosis, expression of Bcl-2 and Bax in rat hepatic stellate cell (HSC-T6).METHODS10% blueberry serum at low, middle and high dose, 10% Fu-Fang-Bie-Jia-Ruan-Gan tablet serum and 10% saline serum were prepared by method of serum pharmacology. Subcultured HSC-T6 was divided into saline serum control group, blueberry serum at low, middle, high dose and Fu-Fang-Bie-Jia-Ruan-Gan tablet serum group, and then was respectively incubated at different dose of 10% blueberry serum, 10% Fu-Fang-Bie-Jia-Ruan-Gan tablet serum and 10% saline serum for 72 hours.Apoptosis of HSC-T6 was detected using flow cytometry with annexin V FITC/PI double staining. The expression of Bcl-2 and Bax in HSC-T6 were examined using immunocytochemistry and Western blotting, respectively.RESULTThere was no significant difference for HSC-T6 Bax protein expression in the low, middle and high dose blueberry serum groups, compared with saline serum control group, respectively.In the high-dose blueberry serum group HSC-T6 early and total apoptosis rate increased significantly compared with the saline serum control group (5.55% ± 0.98% vs 2.53% ± 0.46%, 7.01% ± 1.05% vs 2.96% ± 0.81%, both P<0.05); Bcl-2 protein expression was significantly decreased (A value, 82 ± 35 vs 51 ± 13, P<0.05); Bcl-2/Bax ratio was significantly decreased (0.26 ± 0.02 vs 0.46 ± 0.03, P<0.05); HSC-T6 early and total apoptosis rate, Bcl-2 expression and Bcl-2/Bax ratio in the low and the middle dose blueberry serum group showed no significant difference with the saline serum control group.CONCLUSIONBlueberry can induce HSC-T6 apoptosis by down-regulating Bcl-2 expression and decreasing the ratio of Bcl-2/Bax in HSC-T6 cells, so it may have potential interference effects on hepatic fibrosis.

OBJECTIVETo 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.METHODSForty 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.RESULTSBoth 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.CONCLUSIONDown-regulation of regucalcin and prohibitin gene expression might contribute to the pathogenesis of liver cirrhosis.

OBJECTIVE:To explore the effects of blueberry on the expressions of peroxisome proliferator-activated receptor γ (PPARγ) and platelet-derived growth factor B (PDGF-B) in rat hepatic fibrosis.METHODS:A total of 45 male Sprague-Dawley rats were randomly divided into control group, CCl4-induced hepatic fibrosis (model group), blueberry prevention (BB group), DSHX prevention (DSHX group) and blueberry+DSHX prevention (BB+DSHX group) (n = 9 each). Fibrous liver models of rats were induced by subcutaneous injection of CCl4 and high-lipid/low-protein diet for 8 weeks except for control group. Then the expressions of collagen I (ColI), PPARγ and PDGF-B were determined by real-time polymerase chain reaction (RT-PCR), immunohistochemistry and Western blot respectively.RESULTS:Compared with the control group, the expression of PPARγ decreased and those of ColI and PDGF-B were elevated in model group (P < 0.05). The expression of PPARγ increased and ColI and PDGF-B decreased in BB, DSHX and BB+DSHX groups as compared to model group (P < 0.05).CONCLUSION:The inhibitory effects of blueberry in CCl4-induced liver fibrosis may be correlated with the activation of PPARγ, the inhibited expression of PDGF-B and the reduced synthesis of extracellular matrix.

Objective: To study the effects of blueberry on ultrastructural and expression of cytochrome P450 2E1(CYP2E1) in rats with immune hepatic fibrosis.Methods:Fifty male Wistar rats were randomly divided into normal group,model group,blueberry juice prevention group(blueberry group),Fufang Biejia Ruangan Tablet prevention group(Compounel Group),blueberry juice + Fufang Biejia Ruangan Tablet prevention group(blueberry+Compounel Group).The hepatic fibrosis model of rat was established by porcine serum.Simultaneously,rats in prevention groups were respectively given orally blueberry juice or FFBJRG tablet or combinations of blueberry juice and FFBJRG tablet.All rats were killed at the end of the twelfth week.Ultrastructural changes in hepatocytes was evaluated,expression of hepatic CYP2E1 was detected by real-time reversed transcribed-polymerase chain reaction(RT-PCR),Western blot and immunohistochemical techniques.Results: Compared with model group,the stage of hepatic fibrosis in blueberry group was significantly weakened(P0.05),ultrastructural alterations were markedly attenuated,the expression of CYP2E1 was not significantly different.Conclusion: Blueberry has protective effects on rat immune hepatic fibrosis and alleviates hepatocyte ultrastructural alterations,while it has no effect on the expression of CYP2E1.

OBJECTIVETo observe the effects of blueberry on rat with hepatic fibrosis and ultrastructural. of hepatocytes.METHODSSixty (60) healthy Wistar rats were randomly divided into six groups: normal control group (group A), hepatic fibrosis model group (group B), blueberry at low, middle and high concentration groups (group C, D, E), Fu-Fang-Bie-Jia-Ruan-Gan tablet group (group F). The hepatic fibrosis model of rat was established by intraperitoneal injection of porcine serum once daily for 12 weeks. Simultaneously, rats in groups C-F were respectively perfused with blueberry juice or Fu-Fang-Bie-Jia-Ruan-Gan tablet for 12 weeks except for the normal control group which accepted saline alone. Upon terminal sacrifices of all rats at the end of the twelve weeks. Pathology of hepatic tissue was evaluated by hematoxylin-eosin (HE), Masson staining and transmission electron microscope. Liver index were measured. Levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were examined. Activities of superoxide dismutase (SOD), the contents of malondialdehyde (MDA), hydroxyproline (Hyp) and reduced glutathione (GSH) in liver homogenates were determined.RESULTSRefer to the serum levels of ALT and AST, there is no significant difference existed in all groups (P > 0.05). Activities of SOD, the contents of GSH in liver homogenates of group D and E were significantly higher than those of group B(P < 0.05), while liver index, the contents of Hyp, MDA in liver homogenates were significantly lower than those of group B (P < 0.05). Compared with the group B, the pathological stages of hepatic fibrosis in group D and E were significantly reduced (P < 0.05), the expression of collagen were decreased, ultrastructural alterations were markedly improved.CONCLUSIONBlueberry may have certain preventional protective effects on porcine serum induced rat hepatic fibrosis and exhibit certain protective effects on organelles of hepatocytes (such as mitochondria), especially in middle and high does of blueberry. Its mechanism is probably related to increase the ability of antioxidant stress by increasing SOD activity and GSH content and reducing MDA content.

OBJECTIVE:To study the protective effects of blueberry against rat immune hepatic fibrosis, specifically through the expression of hepatic cytochrome P450 2E1.METHOD: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.RESULTS: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).CONCLUSION: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.

Objective:To investigate the effect of compound glutathione inosine injection(CGII) on the expression of the matrix metalloproteinase-13(MMP-13) and the tissue inhibitor of metalloproteinase-1(TIMP-1) in rats with immunological hepatic fibrosis induced by porcine serum.Methods: Immunological liver fibrosis was induced by intraperitoneal injection of porcine serum for 8 weeks in male Wistar rats.Rats were randomly divided into 6 groups,and treated with 3 doses of CGII and glutathione(GSH,positive control).At the end of the 6th week,hepatic histological changes were observed and scored under optical microscope after HE staining.The mRNA and proteins expressions of MMP-13 and TIMP-1 were detected by real-time RT-PCR and Western blotting.Results: Compared with liver fibrosis control,pathological lesion of liver fibrosis was obviously improved in rats treated by 3 doses of CGII.The mRNA and protein levels of TIMP-1 were significantly reduced but those of MMP-13 were obviously increased in rats treated by 3 doses of CGII as compared with liver fibrosis control.Conclusion: The development of hepatic fibrosis induced by porcine serum in rats is inhibited by CGII.The underlying mechanism might be related with decreasing the expression of TIMP-1 and increasing that of MMP-13.

Objective: To investigate the effect of compound glutathione inosine injection(CGII) on the expression of platelet-derived growth factor-B(PDGF-B) and transforming growth factor-β1(TGF-β1) in rats with immunological hepatic fibrosis.Methods: Immunological hepatic fibrosis was induced by intraperitoneal injection of porcine serum.Then the rats were treated with CGII for 6 weeks.The mRNA and protein expressions of PDGF-B and TGF-β1 in liver tissue were detected by real-time RT-PCR and Western blotting,respectively.Results: Compared with hepatic fibrosis control,the mRNA levels of PDGF-B and TGF-β1 were significantly reduced in rats treated with CGII(5.4,10.8 mg·kg-1,im);the protein levels of PDGF-B and TGF-β1 were significantly reduced in rats treated with CGII(2.7,5.4,10.8 mg·kg-1,im).Conclusion: CGII can inhibit the development of hepatic fibrosis induced by porcine serum in rats.The underlying mechanism might be related with decreasing the expression of PDGF-B and TGF-β1 in the liver.

AIM:To investigate the effects of blueberry on hepatic fibrosis and NF-E2-related factor 2 (Nrf2) transcription factor in rats.METHODS:Forty-five male Sprague-Dawley rats were randomly divided into control group (A); CCl(4)-induced hepatic fibrosis group (B); blueberry prevention group (C); Dan-shao-hua-xian capsule (DSHX) prevention group (D); and blueberry + DSHX prevention group (E). Liver fibrosis was induced in rats by subcutaneous injection of CCl(4) and a high-lipid/low-protein diet for 8 wk (except the control group). The level of hyaluronic acid (HA) and alanine aminotransferase (ALT) in serum was examined. The activity of superoxide dismutase (SOD), glutathione-S-transferase (GST) and malondialdehyde (MDA) in liver homogenates was determined. The degree of hepatic fibrosis was evaluated by hematoxylin and eosin and Masson staining. Expression of Nrf2 and NADPH quinone oxidoreductase 1 (Nqo1) was detected by real-time reversed transcribed-polymerase chain reaction, immunohistochemical techniques, and western blotting.RESULTS:Compared with group B, liver indices, levels of serum HA and ALT of groups C, D and E were reduced (liver indices: 0.038 +/- 0.008, 0.036 +/- 0.007, 0.036 +/- 0.005 vs 0.054 +/- 0.009, P < 0.05; HA: 502.33 +/- 110.57 ng/mL, 524.25 +/- 255.42 ng/mL, 499.25 +/- 198.10 ng/mL vs 828.50 +/- 237.83 ng/mL, P < 0.05; ALT: 149.44 +/- 16.51 U/L, 136.88 +/- 10.07 U/L, 127.38 +/- 11.03 U/L vs 203.25 +/- 31.62 U/L, P < 0.05), and SOD level was significantly higher, but MDA level was lower, in liver homogenates (SOD: 1.36 +/- 0.09 U/mg, 1.42 +/- 0.13 U/mg, 1.50 +/- 0.15 U/mg vs 1.08 +/- 0.19 U/mg, P < 0.05; MDA: 0.294 +/- 0.026 nmol/mg, 0.285 +/- 0.025 nmol/mg, 0.284 +/- 0.028 nmol/mg vs 0.335 +/- 0.056 nmol/mg, P < 0.05). Meanwhile, the stage of hepatic fibrosis was significantly weakened (P < 0.05). Compared with group A, the activity of GST liver homogenates and expression levels of Nrf2 and Nqo1 in group B were elevated (P < 0.05). The expression level of Nrf2 and Nqo1 in groups C, D, and E were increased as compared with group B, but the difference was not significant.CONCLUSION:Blueberry has preventive and protective effects on CCl(4)-induced hepatic fibrosis by reducing hepatocyte injury and lipid peroxidation. However, these effects may not be related to the activation of Nrf2 during long-term of CCl(4).

OBJECTIVE:To investigate the effects of blueberry on the proliferation and activation of hepatic stellate cell (HSC) and its mechanism. METHODS:Rat HSC were isolated by type IV collagenase digestion and Nycodenz density gradient centrifugation. The cultured HSC was incubated at different concentrations of 10% blueberry serum. The 10% DSHX serum was used as positive control and 10% normal rat serum group as control. MTT colorimetric assay was used to detect the HSC proliferation. ColI of culture supernatant was detected by ELISA. The expression of α-SMA in HSC was measured by immunocytochemistry staining while the expressions of Nrf2 and HO-1 were determined by Western blot. RESULTS:Compared with controls, the low and high-dose blueberry serum groups significantly inhibited the HSC proliferation (P < 0.05). It had the same inhibitory effects as the positive control serum group (P > 0.05). ColI of culture supernatant obviously decreased (P < 0.05). And the expression levels of α-SMA in low and high-dose blueberry serum groups decreased significantly (P < 0.05). And there were similar effects in low & high-dose blueberry serum and positive control serum groups. Western blot showed that the expressions of Nrf2 and HO-1 in blueberry and positive control serum groups were higher than that in control group. And the increment was more significant in the low and high-dose blueberry serum groups (P < 0.05). CONCLUSION:Blueberry can significantly inhibit the proliferation and activation of HSC and reduce the synthesis of extra-cellular matrix. It may have potential preventive and protective effects on hepatic fibrosis. The mechanism may be related to an elevated expression of HO-1 through the Nrf2 pathway.

BACKGROUND:Conventional drugs used in the treatment and prevention of liver diseases often have side effects, therefore research into natural substances are of significance. This study examined the effects of blueberry on liver protection and cellular immune functions.METHODS:To determine the effects of blueberry on liver protective function, male mice were orally administered blueberry (0.6 g/10 g) or normal saline for 21 days. Hepatic RNA was extracted by Trizol reagent, and the expression of Nrf2, HO-1, and Nqo1 was determined by real-time RT-PCR. Superoxide dismutase (SOD) and malondialdehyde (MDA) in liver homogenate were determined, and liver index was measured. To assess the effects of blueberry on cellular immune function, male mice received blueberry (0.4, 0.6, or 0.8 g/10 g) for 35 days, and the percentages of CD3+, CD4+, and CD8+ T lymphocyte subgroups in peripheral blood were detected by flow cytometry, the index of the thymus and spleen was measured, and lymphocyte proliferation in the spleen was determined by MTT assay.RESULTS:Blueberry treatment significantly increased the expression of Nrf2, HO-1, and Nqo1, the important antioxidant components in the liver. Hepatic SOD in the blueberry group was higher and MDA was lower than that in the control group (P<0.05). Blueberry also increased the index of the spleen and enhanced the proliferation of lymphocytes of the spleen (P<0.05). The percentages of the CD3+ and CD4+ T lymphocyte subsets and the CD4+/CD8+ ratio were also increased by blueberry (P<0.05).CONCLUSIONS:Blueberry induces expression of Nrf2, HO-1, and Nqo1, which can protect hepatocytes from oxidative stress. In addition, blueberry can modulate T-cell function in mice.

Objective: To study the therapeutic effect of compound glutathione inosine injection(CGII) on liver fibrosis immunologically induced by porcine serum in rats.Methods: The liver fibrosis was induced by intraperitoneal injection of porcine serum.The serum activities of alanine aminotransferase(ALT),aspartate aminotransferase(AST),albumin(A) and globulin(G) were assayed by automatic biochemistry detection instrument.The serum haluronic acid(HA),a hepatic fibrosis index,was determined by ELISA method.Liver index was calculated;hydroxyproline(Hyp) content in liver tissue was assayed by alkaline hydrolysis colorimetry.The hepatic histological changes were also observed by optical microscopy after HE and Masson staining.Results: In rats with liver fibrosis,CGII(2.7,5.4,10.8mg·kg-1,im) significantly decreased the serum levels of AST,ALT,A/G and HA;CGII(5.4,10.8mg·kg-1,im) obviously improved the pathological lesion of liver fibrosis and decreased Hyp contents in liver;CGII(10.8mg·kg-1,im) also decreased liver index.Conclusion: CGII has a therapeutic effect on hepatic fibrosis immunologically induced by porcine serum in rats.

目的:初步探讨过氧化物酶体增殖物激活受体γ（PPARγ）、核转录因子B（NF-κB）的表达与水砷暴露致大鼠肝纤维化的相关性.方法:110只SD大鼠随机分成对照组（自来水）、模型组（浓度100mg/L亚砷酸钠溶液）、自然恢复组（浓度100mg/L亚砷酸钠溶液+自来水）.对照组和模型组分别于第l、2、3、4月末各处死10只,自然恢复组先给予砷溶液,分别在第l、2、3月末取出10只改给予1mo自来水饮用后处死.肝组织病理学检查以观察肝纤维化的动态变化,实时荧光定量RT-PCR法和Western blot法检测PPARγ、NF-κB的mRNA及蛋白表达水平.结果:（1）病理结果:HE染和Masson染色可见,随砷暴露时间的延长,肝细胞变性、坏死增多,汇管区炎症细胞浸润加重,纤维组织增生增多,肝纤维化趋势明显.砷暴露1mo脱离自然恢复1mo后较同月模型组肝细胞变性、坏死及炎细胞浸润程度明显减轻,胶原生成减少.砷暴露2、3mo脱离自然恢复1mo后较同月模型组病理结果差异不明显;（2）mRNA水平:模型组PPARγ mRNA含量逐渐降低,与对照组比较差异均有统计学意义（174.99±41.48,114.55±21.30,64.67±9.83,19.20±16.10 vs 218.40±47.85,P＜0.05）,砷暴露1、2、3mo后分别自然恢复1mo大鼠肝组织中PPARγ mRNA表达均低于同月造模组,仅砷暴露1mo自然恢复组1mo组PPARγ mRNA降低有统计学意义（174.99±41.48 vs 215.97±45.96,P＜0.05）;模型组NF-κB mRNA含量逐渐升高,与对照组比较差异均有统计学意义（65.58±13.17,90.23±15.68,117.95±18.19,172.86±32.92 vs 30.84±15.24,P＜0.05）,砷暴露1、2、3mo后分别自然恢复1mo大鼠肝组织中NF-κB mRNA表达均高于同月造模组,仅砷暴露1mo自然恢复组1mo组NF-κB mRNA升高有统计学意义（65.58±13.17 vs 40.45±19.56,P＜0.05）;（3）蛋白水平:模型组大鼠肝组织中PPARγ的蛋白含量表达均低于对照组,造模3、4mo组与造模1mo组比较差异有统计学意义（0.63±0.06,0.55±0.11 vs 0.85±0.08,P＜0.05）;模型组大鼠肝组织中NF-κB的蛋白含量均高于对照组,造模3、4mo组与造模1mo组比较差异有统计学意义（3.25±0.89,4.27±1.26 vs 1.6±0.57,P＜0.05）;（4）PPARγ和NF-κB的相关性:两者mRNA的表达呈负相关（r=0.847,P＜0.01）,两者蛋白表达也呈负相关（r=0.529,P＜0.05）.结论:肝纤维化程度随砷暴露时间延长而加重,越早脱离砷环境,肝损伤自然恢复越快;砷暴露时间越长,PPARγ mRNA及蛋白表达越低,NF-κB mRNA及蛋白表达越高,二者存在一反馈抑制通路;PPARγ-NF-κB信号传导通路参与砷暴露致肝纤维化形成机制.

OBJECTIVE To study the protective effect of Blueberry against rat hepatic fibrosis and the effect of Blueberry on HO-1 expression patterns. METHODS 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. RESULTS 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. CONCLUSION 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.

OBJECTIVE:To discuss and compare the model establishment of liver fibrosis in oral arsenic solution exposed mice and mice with high-fat feedstuff.METHODS:A total of 240 mice were divided randomly into 6 groups: control group, sodium arsenite group, sodium arsenate group, high-fat feedstuff group, sodium arsenite group with high-fat feedstuff and sodium arsenate group with high-fat feedstuff with 40 mice each. Control group and high-fat feedstuff group (drinking tap water), sodium arsenite group and sodium arsenite group with high-fat feedstuff (drinking 300 mg/L iAs3+ water), sodium arsenate group and sodium arsenate group with high-fat feedstuff (drinking 300 mg/L iAs5+ water). The mice were sacrificed after 3, 6, 10 months' arsenic-exposure and examined for liver function. HE dyeing and Masson dyeing were also employed to observe the pathological changes in hepatic tissue in each group.RESULTS:After 3 months' modeling, ALT and AST in control group, sodium arsenite group, sodium arsenate group, sodium arsenite group with high-fat feedstuff and sodium arsenate group with high-fat feedstuff were (36.7 +/- 5.7) U/L and (110 +/- 22) U/L, (55.6 +/- 4.6) U/L and (249 +/- 41) U/L, (52.6 +/- 8.8) U/L and (161 +/- 15) U/L, (311.3 +/- 19.7) U/L and (484 +/- 15) U/L and (515.0 +/- 60.8) U/L and (671 +/- 24) U/L. They were higher in all the arsenic groups than in control group (P < 0.05); all the HE dyeing samples in arsenic groups showed liver injury in varying degrees such as hydropic degeneration, fatty degeneration, spotty necrosis, focal necrosis and inflammatory cell infiltration. There were liver cell regeneration and fibroplasia in varying degrees. The liver injury of the mice in all arsenic groups aggravated as exposure time prolonged. Masson dyeing after 10 months' modeling showed hyperplasia in portal areas and central venous areas; the mean area of fibrosis in control group, sodium arsenite group, sodium arsenate group, sodium arsenite group with high-fat feedstuff and sodium arsenate group with high-fat feedstuff were 0.1333, 0.5584, 0.5250, 0.7534 and 0.7200 respectively. There was statistical significance between arsenic groups and control group (P < 0.05)CONCLUSION:The liver injury and fibrosis model in oral arsenic solution exposed mice and those with high-fat feedstuff are successfully established and subsequently evaluated. It is a comparatively ideal animal model for studying arsenic liver injury and fibrosis.

<正>长期砷暴露与肝损伤、肝纤维化、肝硬化及肝癌的发生有密切关系,严重危害患者的健康.目前,砷致肝病的研究越来越引起人们的重视,但其发病机制尚未完全阐明,这给临床防治带来较大的困难.对砷与肝损伤相关性及发病机制的深入研究,

血小板衍生生长因子(platelet-derivedgrowth factor,PDGF)对多种细胞均有促增殖和趋化作用,它是HSC最强的有丝分裂原[1].PDGF受体由两种亚单位α及β构成,静止的HSC只表达α亚单位,激活的HSC才表达β亚单位[2],PDGFβ受体在肝纤维化过程中的作用尤为突出.Pinzani等[3]研究表明PDGF-BB能够增加无血清培养的人HSC细胞外信号调节激酶1/2(extracellularsignal regulated kinase 1/2,ERK1/2)的活性,ERK1/2特异性抑制物PD98059能显著抑制PDGF诱导的人HSC活化、增殖[4],这些作用是借助PDGFβ受体实现的.本研究选取PDGF细胞信号转导途径中PDGFR-β和p-ERK1/2两个关键点作为研究对象,以期进一步阐明肝纤维化疾病发生发展的病理生理机制.

AIM: To investigate the effects of Danshao Huaxian capsule (DSHX) on the expression of platelet-derived growth factor receptor β and phosphorylated ERK1/2 (p-ERK1/2) in rats with experimental hepatic fibrosis, and to explore the anti-fibrosis mechanism of DSHX. METHODS: A total of 55 male Sprague Dawley rats were randomly divided into group A, B, and C. The rats in group A served as normal controls, and those in group B and C were treated with carbon tetrachloride, ethanol and high-fat diet to establish the model of liver fibrosis. Meanwhile, the rats in group C were intragastrically administered with 0.8 g/kg DSHX once per day for 8 weeks. HE staining was used to observe the pathological changes of liver tissues. The expression of PDGFR-β and p-ERK1/2 in fibrosis tissues were detected by immunohistochemistry and Western blot, respectively. Biochemical examination were performed to determine the serum levels of hyaluronic acid (HA), laminin (LN), type III procollagen peptide (P IIIP), albumin (ALB), total protein (TP), and the ratio of albumin to globulin (A/G) was also calculated. RESULTS: Pathological examination confirmed that the model of rat hepatic fibrosis was induced successfully at the end of the 8th week. As compared with those in group A, the expression of PDGFR-β, p-ERK1/2 in liver tissues, the serum levels of HA, LN, and PIIIP were significantly increased (PDGFR-β: 184.6 ± 8.5 vs 89.6 ± 5.8, P < 0.05; p-ERK1/2: 360.0 ± 14.5 vs 15.4 ± 2.1, P < 0.05; HA: 517.5 ± 91.5 μg/L vs 254.4 ± 33.1 μg/L, P < 0.05; LN: 58.4 ± 11.3 μg/L vs 37.3 ± 9.8 μg/L, P < 0.05; PIIIP: 36.9 ± 5.6 μg/L vs 4.7 ±1.5 μg/L, P < 0.05) in group B, while the serum level of ALB and the value of A/G were markedly decreased (ALB: 27.4 ± 4.9 g/L vs 42.1 ± 1.6 g/L, P < 0.05; A/G: 0.89 ± 0.08 vs 1.38 ± 0.09, P < 0.05). However, in group C, the expression of PDGFR-β, p-ERK1/2 in liver tissues, the serum levels of HA, LN, and PIIIP were markedly decreased (PDGFR-β: 91.1 ± 6.3 vs 184.6 ± 8.5, P < 0.05; p-ERK1/2: 253.8 ± 18.2 vs 360.0 ± 14.5, P < 0.05; HA: 322.9 ± 41.4 vs 517.5 ± 91.5 μg/L, P < 0.05; LN: 46.0 ± 9.4 vs 58.4 ± 11.3 μg/L, P < 0.05; PIIIP: 14.5 ± 2.4 vs 36.9 ± 5.6 μg/L, P < 0.05), while the serum level of ALB and the value of A/G were notably increased (ALB: 37.2 ± 2.8 g/L vs 27.4 ± 4.9 g/L, P < 0.05; A/G: 1.18 ± 0.13 vs 0.89 ± 0.08, P < 0.05) in comparison with those in group B. CONCLUSION: PDGFR-β and p-ERK1/2 may play pivotal roles in the pathogenesis of liver fibrosis, and DSHX can prevent liver from fibrosis by decreasing the expression of PDGFR-β and p-ERK1/2.

Objective: To report the therapeutic effects of Han-Dan-Gan-Le (HDGL) in the treatment of As-induced liver disorders in humans. Methods: Twenty chronic arsenic-exposed patients were selected based on the history of As exposure, symptoms of liver disorders, and serology excluding HBV and HCV infection. HDGL was administered at the dose of 1.2 g, tid, po for 3 months. Results: After the treatment, all the patients achieved significant improvement of the gastrointestinal tract symptoms, the abdomen pain disappeared, and serum levels of ALT was decreased (P < 0.05). Histology from liver biopsy samples was performed before and at the end of the 3-month treatment. Before treatment, histological examination showed widespread parenchymal cell vacuolation, focal necrosis, inflammatory cell infiltration in focal area and a few foci of fibrosis. After HDGL treatment, hepatocellular vacuolation was greatly ameliorated and focal necrosis and inflammation were not evident, and no liver fibrosis was found. The degree of hepatic histology in this before-group was significantly decreased compared with that in the after- group (P < 0.05). No apparent side effects were observed to stop the HDGL treatment. Conclusions: This preliminary study has demonstrated that HDGL is very effective in the treatment of As-induced liver disorders, as evidenced from histopathology, appetite and other clinical improvements.