自体脾内移植的肝细胞凋亡及与其功能的关系

目的探讨肝细胞脾内移植后发生非免疫排斥性细胞凋亡及其对总体功能发挥的影响。方法应用TUNEL法原位观察80例大鼠肝细胞脾内移植后移植肝细胞的凋亡;分析移植肝细胞凋亡指数与肝化脾ALT含量及同位素99mTC-HIDA摄取量的相关性。结果应用了肝细胞生长因子的大鼠肝细胞脾内移植后移植肝细胞的凋亡指数为（2．76±1．08）,而未应用者则为（5．26±2．14）,两者差异有显著性（P＜0.01）。两组中移植肝细胞凋亡指数与肝化脾ALT含量和99mTc-HIDA摄取量均呈负相关。结论肝细胞脾内移植后较易发生非免疫排斥性细胞凋亡,其凋亡指数与肝细胞生存量及总体功能呈负相关,肝细胞生长因子可抑制移植肝细胞凋亡,从而提高其长期存活量。     

肝细胞移植的新进展

肝细胞移植是继肝移植之后的针对肝功能衰竭的又一有效治疗手段,同时也可以作为急性肝功能衰竭患者等待行肝移植之前的一种桥接治疗手段.与肝移植相比,肝细胞移植具有创伤小、排异反应低等优点,但肝细胞移植同样受到供体短缺的制约.肝细胞移植能否作为一种临床广泛应用的治疗手段主要取决于肝细胞的数量和质量以及合适的移植部位,这方面研究在我国尚未受到充分的重视,因此,本文对肝细胞移植的基础研究与临床应用的新进展作一综述.     

载肝细胞生长因子的纳米粒子治疗急性肝衰竭大鼠

目的 观察载肝细胞生长因子(HGF)的聚乳酸-O-羧甲基壳聚糖(PLA-O-CMC)纳米粒子培养的大鼠肝细胞腹腔内移植对急性肝衰竭(ALF)大鼠的治疗效果.方法 分别将5 ml培养24 h的大鼠肝细胞(5.0×107个)(Ⅰ组)、PLA-O-CMC纳米粒子培养的大鼠肝细胞(Ⅱ组)、载HGF的PLA-O-CMC纳米粒子培养的大鼠肝细胞(Ⅲ组)移植到ALF大鼠腹腔内,以PLA-O-CMC纳米粒子培养的大鼠肝细胞腹腔移植加每天静脉注射HGF 10 μg/kg×7 d(Ⅳ组)和5 ml RPMI 1640培养基腹腔内注射(Ⅴ组)作为对照.观察受体大鼠存活率、肝功能、肝组织光镜和电镜变化.结果 移植后14d大鼠的存活率Ⅰ～Ⅴ组分别为50.00％、68.75％、81.25％、75.00％和18.75％,各移植组高于对照组,Ⅲ组最高.移植后24 h,Ⅱ～Ⅳ组各项肝功能指标开始好转,至移植后7d,各组间除谷丙转氨酶(ALT)外,差异无统计学意义(P＞0.05).Ⅲ组的肝功能和肝脏病理损害恢复最快,其次为Ⅳ、Ⅱ、Ⅰ组,Ⅴ组恢复最慢、最差.结论 应用载HGF的PLA-O-CMC纳米粒子培养的大鼠肝细胞腹腔内移植治疗ALF大鼠能逆转肝功能,提高生存率,较静脉途径给予HGF的治疗效果更好.     

nodosin促进大鼠部分肝移植肝细胞再生的实验研究

目的  探讨溪黄草有效成分nodosin对大鼠减体积肝移植术后肝细胞再生的影响。方法  以Wistar大鼠作为供体, SD大鼠作为受体, 采用改良的二袖套法建立大鼠部分肝移植模型, 将24只受体大鼠随机分为nodosin组和对照组。nodosin组于肝移植术后经尾静脉注射nodosin 100μg/ml。在术后3 d和7 d分别处死大鼠取外周血浆及肝脏组织标本。分光光度法检测外周血浆中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和白蛋白(ALB)含量; 光学显微镜下观察肝组织形态学改变; 采用免疫组织化学法检测肝组织增殖细胞核抗原(PCNA)抗体的表达水平; 采用蛋白印迹法(Western blot)检测肝组织中磷酸化蛋白激酶(p-AKT)、磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)、细胞周期素D1(cyclin D1)、血红素加氧酶(HO)-1蛋白的表达水平; 采用磷脂酰丝氨酸外翻分析(Annexin V法)和dUTP缺口末端标记(TUNEL)法检测肝细胞的凋亡情况。结果  术后3 d和7 d, 与对照组比较, nodosin组大鼠血清中ALT、AST含量较低, ALB含量较高(均为P < 0.05), 同时nodosin能够减轻大鼠肝移植术后肝组织病理损伤。nodosin组的PCNA阳性表达细胞数明显多于对照组(P < 0.05)。nodosin组增殖相关蛋白p-AKT、p-mTOR、cyclin D1和HO-1蛋白表达水平均明显高于对照组(均为P < 0.05)。nodosin组发生凋亡的肝细胞数量和比例明显低于对照组(P < 0.05)。结论  大鼠肝移植术后应用nodosin具有减少肝细胞凋亡数量, 促进肝细胞增殖的作用。     

急性肝衰竭大鼠载肝细胞生长因子纳米粒子肝细胞移植后有丝分裂指数和Ki-67表达变化及意义

目的 探讨急性肝衰竭(ALF)大鼠载肝细胞生长因子(HGF)的聚乳酸-氧-羧甲基壳聚糖(PLA-O-CMC)纳米粒子肝细胞移植后有丝分裂指数和Ki-67抗原的表达变化及意义.方法 D-氨基半乳糖腹腔内注射制作大鼠ALF模型,48 h后分别将Ⅰ型胶原(Ⅰ组)、PLA-O-CMC纳米粒子(Ⅱ、Ⅳ组)、载HGF的PLA-O-CMC纳米粒子(Ⅲ组)培养的大鼠肝细胞(均为5×107个,5 ml)移植到ALF大鼠腹腔内.以每日静脉注射HGF 10μg/kg×7 d(Ⅳ组)、5 Ml RPMI 1640腹腔内注射(Ⅴ组)作为对照.观察其14 d存活率、血清及肝组织HGF浓度、肝组织病理及有丝分裂指数(MI)、Ki-67变化.结果移植后14 d Ⅰ～Ⅴ组ALF大鼠的存活率分别为50.00％、68.75％、81.25％、75.00％、18.75％.各纳米移植组高于V组.Ⅲ组3 d时肝组织HGF浓度最高,平均为5882.91 μG／L.Ⅲ组肝组织结构恢复更快,5 d时平均MI 10.20％0、7 d时平均MI 10.20‰、7 d时Ki-67平均标记指数16.8‰,均高于Ⅴ组.结论 应用载HGF的PLA-O-CMC纳米粒子培养的大鼠肝细胞腹腔内移植治疗ALF大鼠能促进肝再生相关抗原表达.

Abstract：

Objective To evaluate the change and significance of Ki-67 antigen expression follow ing hepatocyte transplantation using hepatocyte growth factor (HGF) loaded polylactic acid-O-carboxymethylchitosan (PLA-O-CMC) nanoparticles in rats with acute liver failure (ALF). Methods ALF models of rats were established by D-galactosamine intraperitoneal injection. Rat hepatocytes type Ⅰ collagen suspension (group Ⅰ ),rat hepatocytes co-cultured with PLA-O-CMC nanoparticles ( groups Ⅱ ,Ⅳ ) and rat hepatocytes co-cultured with HGF loaded PLA-O-CMC nanoparticles ( group Ⅱ ) (5 × 107 cells each group,5 ml) were transplanted into the abdominal cavity of SD rats at 48 h after D-Gal injection. Intravenous injection of HGF ( 10 μg/kg for 7 days) ( group Ⅳ ) and RPMI 1640 injection (group Ⅴ ) were used as the negative groups. The 14th-day survival rate of rats,pathological change and mitotic index of liver,Ki-67 antigen labeling index of ALF rat livers were observed. Results The 14th-day survival rate in groups Ⅰ -Ⅴ was 50.00％,68. 75％,81.25％,75.00％,18.75％,and that in nano-transplanted groups was higher than in group Ⅴ. At the 3rd day,the concentration of HGF in liver tissue of group Ⅲ,average 5882. 91 μg/L was the highest. The hepatic lobules structure in group Ⅲ recovered faster. The average mitotic index in group Ⅱ at the 5th day was 10. 20‰ and the average Ki-67 labeling index at the 7th day was 16. 8‰,which were all higher than in group Ⅴ. Conclusion Intraperitoneal transplantation of HGF loaded PLA-O-CMC nanoparticle-attached hepatocytes for treatment of ALF can promote the liver regenerationassociated antigen expression.     

Three different hepatocyte transplantation techniques for enzyme deficiency disease and acute hepatic failure

The effects of three different techniques of hepatocyte transplantation were investigated: transplantation of free hepatocytes into the spleen and intraperitoneal transplantation of microcarrier-attached hepatocytes or of microencapsulated hepatocytes. The liver-supportive functions of these transplanted hepatocytes were analyzed using either the Gunn rat (hyperbilirubinemia) or rats with acute liver failure. In the Gunn rat intraperitoneal transplantation of microcarrier-attached hepatocytes resulted in a significant reduction of plasma bilirubin for 28 days whereas intraperitoneal transplantation of microencapsulated hepatocytes was ineffective notwithstanding immunosuppression by cyclosporin A. Intrasplenic hepatocyte transplantation was only effective in reducing plasma bilirubin for 14 days. During acute liver failure, liver support was achieved temporarily by hepatocyte transplantation in the spleen, by intraperitoneally transplanted microcarrier-attached hepatocytes, and by microencapsulated hepatocytes to equal extents, the microencapsulated hepatocytes being the least effective after 8 h of liver ischemia.     

应用环孢素A治疗异种肝细胞移植的排斥反应

目的　探讨应用环孢素A(CsA)治疗异种肝细胞移植排斥反应的疗效及其排斥机制。方法　将豚鼠肝细胞植入D-氨基半乳糖诱导的肝衰大鼠脾内,分CsA治疗组和单纯移植组。观察2周生存率及脾病理组织学检查;酶联免疫双抗体夹心法(Sandwich-ELISA)测定大鼠血清白细胞介素(IL)-2的浓度。结果　2周生存率比较CsA治疗组（78.6%）与单纯移植组（80.0%）相比差异无显著性(P＞0.05)。应用CsA可减缓炎症细胞浸润。血清IL-2浓度检测正常大鼠为(28.7±7.0)ng/L,移植后各组血清IL-2浓度变化不大,直至移植后第7天,CsA治疗组为(28.5±6.0)ng/L、单纯移植组为(31.5±8.0)ng/L。3组间差异均无显著性(P＞0.05)。结论　在异种肝细胞脾内移植中,细胞免疫发生的较迟和/或较弱。单独应用CsA治疗异种肝细胞脾内移植引发的排斥反应其疗效不佳。     

双层法氧合冷保存心跳停搏大鼠肝细胞移植研究

目的 观察双层法(TLM)氧合冷保存较UW保存能否改善心跳停搏供体(NHBD)肝细胞存活率和功能.方法 SD大鼠为供体,建立NHBD模型,NAPs大鼠为受体.根据热缺血时间(WIT)15 m/n和30 m/n分成2组;按TLM、UW分别保存3、12 h和未保存再各分5个亚组(n=5).检测NHBD肝细胞存活率和ATP水平,观察肝细胞移植(HTx)后肝细胞形态和功能.结果 TLM3、12 h组肝细胞存活率分别显著高于UW 3、12 h组[(69.7±4.1)%和(69.1±2.0)%比(55.1±2.3)%和(53.3±2.0)%;P<0.01];TLM 3、12 h组AlP水平分别显著高于UW 3、12 h组(3.25±0.79和3.06±0.67比2.25±0.53和1.63±0.40;P<0.05或P<0.01).HTx后几乎所有时间点TLM组血清白蛋白(ALB)水平都显著高于UW组(P<0.05或P<0.01).在HTx 14d后,形态学显示TLM组肝细胞保持强活力,糖原和ALB染色呈强阳性.结论 TLM氧合冷保存可显著改善和逆转NHBD肝细胞存活率和功能,减少NHBD肝细胞缺血性损伤.     

In vitro study of multicellular hepatocyte spheroids formed in microcapsules

To make highly differentiated encapsulated hepatocytes, we formed hepatocyte spheroids in microcapsules in cultures. Hepatocytes isolated from rats were encapsulated in alginate-poly-L-lysine artificial cells and cultured under different medium conditions. When high molecular weight poly-L-lysine was used to make the capsule membrane, the hepatocytes aggregated and formed spheroids inside microcapsules within 48 hs. We studied the viability and function of encapsulated hepatocyte spheroids; free hepatocyte spheroids; nonaggregated encapsulated hepatocytes; and free hepatocyte monolayers. Cell viability and protein-producing ability were monitored for up to 30 days. Hepatocyte spheroids in the encapsulated or free form remained viable and continued to secrete proteins throughout the 30 days of observation. The viability and function of nonaggregated hepatocytes in the encapsulated or free form declined rapidly. These results suggest that the tridimensional structure of the spheroids may be important in maintaining the viability and function of encapsulated hepatocytes.     

Acute impairment of hepatic microcirculation and recruitment of nonparenchymal cells by intrasplenic hepatocyte transplantation

Background/Purpose

Over the last 20 years, hepatocyte transplantation (HcTx) has advanced from the experimental to the clinical stage. To date, HcTx has been performed in 30 patients in the United States. Regardless whether hepatocytes are transplanted into the spleen and migrate to the liver or are injected directly into the portal vein, transplanted liver cells will, to some extent, congest the recipient liver microcirculation. The potential negative consequences of intrasplenic HcTx were the subject of this study.

Methods

By using intravital microscopy, the authors investigated whether intrasplenic HcTx of 20 × 10⁶ allogenic hepatocytes would influence liver perfusion, excretory liver function, and nonparenchymal cells (Kupffer and Ito cells) in vivo.

Results

The sinusoidal perfusion rate declined significantly from 94% (control) to 84% on day 1 and 76% on day 7. Bile acid excretion decreased in a similar fashion from 0.924 mg/h (control) to 0.669 mg/h on day 7. The authors observed a significant increase of Ito cells from 81.1 cells per microscopic field (control) to 97.1 (day 1) and an increase of Kupffer cells (KC; 6.1 cells per microscopic field on day 1 v 3.8 on control).

Conclusions

This study shows an acute impairment of hepatic microcirculation and hepatucellular function along with an recruitment and activation of nonparenchymal cells in the early posttransplantation period after intrasplenic HcTx. Kupffer cell recruitment indicates an activation of local host defense, and Ito cell activation implies the initiation of liver repair mechanisms owing to ischemia-related cell damage.     

DiI荧光示踪剂在大鼠脾内肝细胞移植中的研究

目的 探讨大鼠肝细胞在脾内移植后的生存、迁移及其并发症情况。方法 先用ＤｉＩ荧光示踪剂标记肝细胞再行脾内肝细胞移植。在不同时间段取肝、脾、肺、心、肾 、胸腺组织冷冻切片,在荧光显微镜下用荧光和普通光示踪观察。结果 ＤｉＩ荧光示踪剂能很好地标记肝细胞,６０ｄ实验期内,移植细胞能在所取各器官中生存并保持良好形态,未发现有区域梗塞灶、同时还反映了其在肝实质中的分布。结论 ＤｉＩ是一种新颖、使用简便的荧光     

肝细胞生长因子基因修饰骨髓间质干细胞移植对大鼠肺内血管生成的影响

目的 评价人肝细胞生长因子(hHGF)基因修饰骨髓间质干细胞(MSCs)移植对大鼠肺内血管生成的影响.方法 F344大鼠20只,体重200～ 250 g,2月龄,采用随机数字表法,将其随机分为2组(n=10),HGF组经颈外静脉注射5× 105个HGF基因修饰MSCs,对照组(C组)给予等容量1ml DMEM培养液.移植后28d,测定平均肺动脉压,然后取肺组织,测定hHGF含量,采用免疫组织化学法检测增殖细胞核抗原表达,以反映肺小血管内皮增生程度,采用免疫组织化学法检测Ⅷ因子表达,以反映肺小血管密度,光学显微镜下观察肺血管病理学结果.结果 与C组比较,HGF组平均肺动脉压差异无统计学意义(P＞0.05),肺组织hHGF含量、小血管内皮增生程度和小血管密度升高(P＜0.01).HGF组肺小血管结构未见异常.结论 hHGF基因修饰的MSCs移植可促进大鼠肺内血管生成.     

Long-term effects of intrasplenically transplanted adult hepatocytes and fetal liver in hyperbilirubinemic Gunn rats

We performed adult hepatocyte transplantation (HCTx) and fetal liver transplantation (FLTx) into the spleens of hyperbilirubinemic Gunn rats in congenic combination and we compared the long-term effects of these procedures for as long as 12 months. Proliferative activity of intrasplenic hepatocytes was evaluated using antiproliferating cell nuclear antigen (PCNA) immunohistochemical staining. The serum total bilirubin levels (T. Bil) significantly decreased from 7.16±0.25 mg/dl to 4.38±0.60 mg/dl 2 months after HCTx and gradually decreased thereafter until 12 months after transplantation (3.23±0.37 mg/dl, P<0.05 vs preoperative value). The T. Bil change after FLTx was similar to that of HCTx: 7.22±0.24 mg/dl before FLTx, and 4.92±0.24 and 3.06±0.47 mg/dl, 2 and 12 months after FLTx (P<0.05), respectively. Bilirubin glucuronides, which were not detectable in the bile from untreated Gunn rats, appeared in considerable amounts 4 months after HCTx and FLTx (27.5% and 36.0% of total bile, respectively). PCNA labeling indices of intrasplenic hepatocytes (4.9%±0.9% and 3.7%±0.7%, 6 months after HCTx and FLTx, respectively) were slightly higher than those of normal hepatocytes (1.0%±0.1%) in the host liver. In conclusion, both adult and fetal rat hepatocytes transplanted into the spleen in congenic combination functioned for at least a year in terms of bilirubin glucuronidation. The spleen is considered to be one of the optimal grafting sites for hepatocytes, with nearly lifelong significant function and proliferative activity.     

肝细胞体内移植治疗重型肝炎临床研究

目的探讨肝细胞体内移植治疗重型肝炎临床应用的安全性、疗效及免疫抑制剂方案的可行性。方法采用肝细胞分离纯化、体外培养、冷冻及复苏方法，经股动脉插管原位肝细胞体内移植。结果7例重型肝炎病人中，2例临床治愈；2例好转出院；1例肝细胞移植后第8天等到供体肝脏行原位肝移植；2例死亡。结论肝细胞体内移植是一项安全，近期疗效确定的治疗方法，免疫抑制剂方案有可行性，移植后的肝细胞能够在脾脏内增殖、分化，替代或部分恢复肝脏合成、解毒和代谢功能。     

免疫隔离技术在异种肝细胞移植中的应用

目的探讨微囊化猪肝细胞腹腔内移植对药物性肝衰大鼠的治疗作用，观察移植大鼠存活率、肝功能的变化。方法以海藻酸钠体外包裹经胶原酶灌注法分离的乳猪肝细胞，以SD大鼠为受体，D-氨基半乳糖腹腔内注射诱导大鼠急性肝衰竭，48h后将微囊化的猪肝细胞移植于大鼠腹腔内，观察移植大鼠存活率、绘制生存曲线，并测定肝功能的变化。结果腹腔内肝细胞移植可显著改善大鼠肝功能，转氨酶及胆红素均较对照组明显下降（P〈0．05）。与裸肝细胞移植组相比．微囊化肝细胞移植组1周存活率（78．6％ vs 66．7％）及2周存活率（42．9％ vs 25．0％）均显著提高（P〈0．01）。结论对异种肝细胞经微囊化处理后移植治疗急性肝衰大鼠，可给予肝功能代谢支持，提高移植治疗效果。     

异种肝细胞移植治疗大鼠药物性肝衰的疗效观察

目的　观察异种肝细胞移植治疗大鼠药物性肝衰的疗效。方法　杂种豚鼠为供体 ,胶原酶消化法制备肝细胞。SD大鼠为受体 ,氨基半乳糖 (D GI)腹腔内 1次注射制作肝衰模型。 4 8h后将豚鼠肝细胞(1.5× 10 7个 )移植于大鼠脾内。同种移植及生理盐水为对照。移植后观察受体 2周存活率 ,在移植不同时间作移植物病理及组织化学检查。结果　受体 2周存活率 :异种移植组为 71% ,同种组为69 % (P >0 .0 5 ) ,生理盐水对照组为 2 5 % (P>0 .0 1)。豚鼠肝细胞移植后 12～ 2 4h其结构和功能基本保存完好。结论　与同种移植一样 ,异种肝细胞移植能逆转大鼠药物性肝衰。     

Hepatocyte transplantation for total liver repopulation

Hepatocyte transplantation (HT) is an attractive therapeutic alternative to liver transplantation. A number of experiments have shown the feasibility of total liver parenchymal cell replacement by transplanted hepatocytes. In this review, we would like to highlight researches and clinical reports of HT for liver repopulation. Cellular source of clinical HT should be safety. Immortalized cells, hepatic stem cells, and other stem cells have been used for an experimental model for HT. The exact mechanism of the cell engraftment after HT has not been completely understood, although there were some markers to detect and investigate transplanted cells. In order to achieve liver repopulation following HT, a mild hepatic damage may need to facilitate cell engraftment and replace the host liver by transplanted cells. Hormonal factor may use for the same purpose. Despite the results of preclinical studies promising clinical benefits for cell therapy, the clinical experience of HT has been disappointing, except in a few cases. HT may become an alternative for liver transplantation in the future; however, many efforts should made before establishing an effective method for HT and liver replacement therapy.     

大鼠肝细胞肾脏内移植后组织形态与肝功能衰竭治疗作用研究

本实验通过同种异体大鼠肝细胞肾脏内移植，尝试在受体体内建立第二肝脏。对移植肝细胞进行长期组织形态与功能观察，并利用该方法进行急性肝功能衰竭肝功能辅助支持治疗研究。结果表明肝细胞肾脏内移植后可长期存活、增殖而且具有肝组织结构重建及保持肝脏功能的能力。４×１０７个肝细胞肾脏内移植后可显著提高受体大鼠对肝功能衰竭的承受力，移植组术后３天行９０％肝切除诱发急性外科型肝功能衰竭后其存活率显著高于对照组。以上结果显示肾脏可以成为肝细胞移植又一适宜部位，肝细胞肾脏内移植有可能成为临床上肝脏代谢及功能障碍疾病的一种治疗手段。     

Increased apoptosis of hepatocytes in vascular occlusion after orthotopic liver transplantation

Early vascular occlusion is liable to cause graft failure, and differential diagnosis between this condition and primary nonfunction (PNF) caused by preservation injury may be difficult. Apoptosis has been detected in immunomediated cytotoxicity and is known to be triggered by mild ischemia. In a retrospective analysis we investigated the role of apoptosis in vascular occlusion, PNF, and acute allograft rejection to improve the differential diagnosis of early graft failure. The liver graft histology of 75 patients (46 male, 29 female) a median 47 (1–64) years of age was screened semiquantitatively for the rate of apoptosis on the hematoxylin-eosin stain (HE) and by the in situ end nick labeling technique (TUNEL). This cohort included all patients who developed PNF (n = 9) or vascular occlusion (n = 11) after orthotopic liver transplantation (OLT) in the years 1992 to 1996. Within this period of time we performed 205 OLTs on 189 patients. We further included 22 patients with early acute rejection and 11 controls. The highest rates of apoptotic hepatocytes were seen in vascular occlusion (P < 0.001). Grafts with PNF were explanted 1–3 days after OLT and showed hepatocytes that were 100 % necrotic. Cases of acute early rejection showed a significantly higher apoptotic cell count than did normal controls (P < 0.003), increasing in direct proportion to the severity of rejection. Screening biopsies for the rate of apoptosis can improve the efficacy and accuracy of differential diagnosis of early graft failure. Received: 27 January 1999/Revised: 7 September 1999/Accepted: 13 September 1999     

转染HGF基因对肝细胞的生物学效应

目的探讨肝细胞生长因子(hepatocyte growth factor，HGF)基因对肝细胞生长增殖能力的影响。方法通过脂质体介导法，将HGF基因导入肝细胞中。用荧光显微镜以及原位杂交观察到HGF基因表达。采用检测细胞生长曲线、Ki-67蛋白和嗜银蛋白免疫组化观察肝细胞生长增殖能力及DNA合成能力的变化。结果荧光显微镜观察可见到绿色荧光蛋白的表达，用原位杂交方法进一步证实了HGF蛋白在细胞中的表达。细胞生长曲线显示，转染HGF基因的肝细胞增殖速度明显增快，Ki-67蛋白和嗜银蛋白表达明显增多，提示转导HGF基因使肝细胞增殖活性增加。结论本实验显示转染的HGF可表达并有促细胞分裂活性。为进一步了解HGF分子生物学特性及治疗应用提供了理论基础。