Taking advantage of the potential of mesenchymal stromal cells in liver regeneration:Cells and extracellular vesicles as therapeutic strategies

Cell-based therapies for acute and chronic liver diseases are under continuous progress. Mesenchymal stem/stromal cells(MSCs) are multipotent cells able to migrate selectively to damaged tissue and contribute to its healing and regeneration. The MSC pro-regenerative effect occurs due to their immunomodulatory capacity and their ability to produce factors that promote cell protection and survival. Likewise,it has been observed that part of their paracrine effect is mediated by MSC-derived extracellular vesicles(EVs). EVs contain proteins,lipids and nucleic acids(DNA,m RNA,mi RNA,lnc RNA) from the cell of origin,allowing for intercellular communication. Recently,different studies have demonstrated that MSC-derived EVs could reproduce,at least in part,the biological effects obtained by MSCbased therapies. Moreover,due to EVs' stability for long periods of time and easy isolation methods they have become a therapeutic option to MSCs treatments. This review summarizes the latest results achieved in clinical trials using MSCs as cell therapy for liver regeneration,the role of EVs in liver physiopathology and the potential of MSC-derived EVs as intercellular mediators and therapeutic tools in liver diseases.

Adipose-derived stromal cell in regenerative medicine:A review

The application of appropriate cell origin for utilizing inregenerative medicine is the major issue. Various kinds of stem cells have been used for the tissue engineering and regenerative medicine. Such as, several stromal cells have been employed as treat option for regenerative medicine. For example, human bone marrow-derived stromal cells and adipose-derived stromal cells(ADSCs) are used in cell-based therapy. Data relating to the stem cell therapy and processes associated with ADSC has developed remarkably in the past 10 years. As medical options, both the stromal vascular and ADSC suggests good opportunity as marvelous cell-based therapeutics. The some biological features are the main factors that impact the regenerative activity of ADSCs, including the modulation of the cellular immune system properties and secretion of bioactive proteins such as cytokines, chemokines and growth factors, as well as their intrinsic anti-ulcer and anti-inflammatory potential. A variety of diseases have been treated by ADSCs, and it is not surprising that there has been great interest in the possibility that ADSCs might be used as therapeutic strategy to improve a wider range of diseases. This is especially important when it is remembered that routine therapeutic methods are not completely effective in treat of diseases. Here, it was discuss about applications of ADSC to colitis, liver failure, diabetes mellitus, multiple sclerosis, orthopaedic disorders, hair loss, fertility problems, and salivary gland damage.

Yinchenhao decoction attenuates obstructive jaundice-induced liver injury and hepatocyte apoptosis by suppressing protein kinase RNA-like endoplasmic reticulum kinase-induced pathway

BACKGROUND Chronic biliary obstruction results in ischemia and hypoxia of hepatocytes,and leads to apoptosis.Apoptosis is very important in regulating the homeostasis of the hepatobiliary system.Endoplasmic reticulum(ER)stress is one of the signaling pathways that induce apoptosis.Moreover,the protein kinase RNA-like endoplasmic reticulum kinase(PERK)-induced apoptotic pathway is the main way;but its role in liver injury remains unclear.Yinchenhao decoction(YCHD)is a traditional Chinese medicine formula that alleviates liver injury and apoptosis,yet its mechanism is unknown.We undertook this study to investigate the effects of YCHD on the expression of ER stress proteins and hepatocyte apoptosis in rats with obstructive jaundice(OJ).AIM To investigate whether YCHD can attenuate OJ-induced liver injury and hepatocyte apoptosis by inhibiting the PERK-CCAAT/enhancer-binding protein homologous protein(CHOP)-growth arrest and DNA damage-inducible protein 34(GADD34)pathway and B cell lymphoma/leukemia-2 related X protein(Bax)/B cell lymphoma/leukemia-2(Bcl-2)ratio.METHODS For in vivo experiments,30 rats were divided into three groups:control group,OJ model group,and YCHD-treated group.Blood was collected to detect the indicators of liver function,and liver tissues were used for histological analysis.For in vitro experiments,30 rats were divided into three groups:G1,G2,and G3.The rats in group G1 had their bile duct exposed without ligation,the rats in group G2 underwent total bile duct ligation,and the rats in group G3 were given a gavage of YCHD.According to the serum pharmacology,serum was extracted and centrifuged from the rat blood to cultivate the BRL-3A cells.Terminal deoxynucleotidyl transferase mediated dUTP nick end-labelling(TUNEL)assay was used to detect BRL-3A hepatocyte apoptosis.Alanine aminotransferase(ALT)and aspartate transaminase(AST)levels in the medium were detected.Western blot and quantitative real-time polymerase chain reaction(qRT-PCR)analyses were used to detect protein and gene expression levels of PERK,CHOP,GADD34,Bax,and Bcl-2 in the liver tissues and BRL-3A cells.RESULTS Biochemical assays and haematoxylin and eosin staining suggested severe liver function injury and liver tissue structure damage in the OJ model group.The TUNEL assay showed that massive BRL-3A rat hepatocyte apoptosis was induced by OJ.Elevated ALT and AST levels in the medium also demonstrated that hepatocytes could be destroyed by OJ.Western blot or qRT-PCR analyses showed that the protein and mRNA expression levels of PERK,CHOP,and GADD34 were significantly increased both in the rat liver tissue and BRL-3A rat hepatocytes by OJ.The Bax and Bcl-2 levels were increased,and the Bax/Bcl-2 ratio was also increased.When YCHD was used,the PERK,CHOP,GADD34,and Bax levels quickly decreased,while the Bcl-2 levels increased,and the Bax/Bcl-2 ratio decreased.CONCLUSION OJ-induced liver injury and hepatocyte apoptosis are associated with the activation of the PERK-CHOP-GADD34 pathway and increased Bax/Bcl-2 ratio.YCHD can attenuate these changes.

Apoptosis induced by NaYF_4:Eu^(3+) nanoparticles in liver cells via mitochondria damage dependent pathway

As lanthanide-doped sodium yttrium flouride(NaYF_4)nanoparticles have great potential inbiomedical applications,their biosafety is important and has attracted significant attention.In the present work,three different sized NaYF_4:Eu^(3+)nanoparticles have been prepared.Liver BRL 3 A cell was used as a cell model to evaluate their biological effects.Cell viability and apoptosis assays were used to confirm the cytotoxicity induced by NaYF_4:Eu^(3+)NPs.Apart from the elevated malondialdehyde(MDA),the decrease of superoxide dismutase(SOD),glutathione peroxidase(GSH-PX)and catalase(CAT)activity indicated reactive oxygen species(ROS)generation,which were associated with oxidative damage.The decrease of mitochondrial membrane potential(MMP)value demonstrated the occurrence of mitochondria damage.Then,release of cytochrome c from mitochondria and activation of caspase-3 confirmed that NaYF_4:Eu^(3+)NPs induced apoptosis was mitochondria damage-dependent.

枸杞多糖对Hcy诱导的小鼠肝细胞损伤的保护作用及其机制

目的探讨枸杞多糖(LBP)对同型半胱氨酸(Hcy)诱导的小鼠肝细胞损伤的作用及其机制。方法体外培养正常C3H/An小鼠肝细胞(NCTC 1469),采用不同浓度的Hcy(0、50、100、200、500μmol/L)处理NCTC 1469细胞,使用MTT法检测不同浓度Hcy对细胞活性的影响。取对数生长期细胞,设置:(1)对照组(采用10%马血清的DMEM培养基培养)与Hcy组(采用100μmol/L Hcy溶液处理48 h),收集细胞。采用细胞活力染色检测细胞凋亡情况,谷草转氨酶(AST)/谷丙转氨酶(ALT)活性检测试剂盒检测AST、ALT活性,RT-qPCR检测YAP1(Yes相关蛋白1)、DNMT1、DNMT3a、DNMT3b mRAN的表达,Western blotting检测YAP1蛋白的表达,巢式甲基化特异性PCR(nMS-PCR)检测YAP1启动子区DNA甲基化率。(2)对照组、LBP组、Hcy组与Hcy+LBP组,LBP组采用4 mg/ml LBP溶液处理2 h,Hcy组、Hcy+LBP采用100μmol/L Hcy溶液处理48 h,46 h时Hcy+LBP组加入4 mg/ml LBP溶液处理,收集细胞。采用RT-qPCR检测YAP1、DNMT1、DNMT3a、DNMT3b mRAN的表达,Western blotting检测YAP1、Bax、Bcl-2蛋白的表达,AST/ALT活性检测试剂盒检测AST、ALT活性。采用生物信息学方法预测YAP1启动子区DNA甲基化CpG岛。结果根据MTT法检测结果,选择100μmol/L Hcy处理NCTC 1469细胞进行后续实验。与对照组比较,Hcy组细胞凋亡率增高(P<0.01),ALT、AST活性增高(P<0.001),YAP1 mRAN和蛋白相对表达水平降低(P<0.001),YAP1启动子区域甲基化率增高(P<0.01),DNA甲基化酶DNMT1、DNMT3a及DNMT3b mRNA相对表达水平增高(P<0.01或P<0.001)。与Hcy组比较,Hcy+LBP组DNA甲基化酶DNMT1、DNMT3a及DNMT3b mRNA相对表达水平降低(P<0.001),YAP1 mRAN和蛋白相对表达水平明显增高(P<0.01或P<0.001),Bcl-2蛋白相对表达水平明显升高(P<0.001),Bax蛋白相对表达水平明显降低(P<0.001),ALT、AST活性降低(P<0.001)。结论YAP1表达降低可能是Hcy诱导的小鼠NCTC 1469细胞损伤的关键过程,YAP1启动子区域甲基化的改变可能是Hcy引起YAP1表达变化的分子机制。LBP可能通过正向调节YAP1的表达改善Hcy引起的小鼠NCTC 1469细胞损伤。

血清SIGIRR、sTRAIL与传染性单核细胞增多症患儿外周血T细胞亚群和肝脏损伤的关系研究

目的 探讨传染性单核细胞增多症(infectious mononucleosis, IM)患儿血清单免疫球蛋白白细胞介素1相关受体(single immunoglobulin interleukin-1 related receptor, SIGIRR)、可溶性肿瘤坏死因子相关的凋亡诱导配体(soluble tumor necrosis factor-related apoptosis-inducing ligand, sTRAIL)与外周血T细胞亚群和肝脏损伤的关系。方法 选取2020-01/2023-01月作者医院收治的95例IM患儿为IM组,另选取同期100例体检健康儿童为对照组。根据IM患儿是否发生肝脏损伤分为肝脏损伤组(n=49)和无肝脏损伤组(n=46)。采用酶联免疫吸附试验法检测血清SIGIRR、sTRAIL水平,流式细胞术检测外周血T细胞亚群(CD3^(+)、CD16^(+)、CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+))。通过多因素Logistic回归分析探讨IM患儿肝脏损伤的影响因素,受试者工作特征(receiver operating characteristic, ROC)曲线分析血清SIGIRR、sTRAIL对IM患儿肝脏损伤的预测价值。结果 与对照组健康儿童比较,IM组患儿血清SIGIRR水平和外周血CD16^(+)、CD4^(+)、CD4^(+)/CD8^(+)降低,血清sTRAIL水平和外周血CD3^(+)、CD8^(+)升高(P均<0.05)。Pearson相关性分析显示,IM患儿血清SIGIRR水平与外周血CD16^(+)、CD4^(+)、CD4^(+)/CD8^(+)呈正相关关系,与CD3^(+)、CD8^(+)呈负相关关系(P<0.05),血清sTRAIL水平与外周血CD16^(+)、CD4^(+)、CD4^(+)/CD8^(+)呈负相关关系,与CD3^(+)、CD8^(+)呈正相关关系(P<0.05)。单因素分析结果显示,病程、白细胞计数、CD3^(+)、CD16^(+)、CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+)、SIGIRR水平、sTRAIL水平为IM患儿肝脏损伤的影响因素(P均<0.05)。多因素Logistic回归分析结果显示,CD3^(+)、CD16^(+)、sTRAIL升高为IM患儿肝脏损伤的独立危险因素,CD4^(+)/CD8^(+)、SIGIRR升高为其保护因素(P<0.05)。ROC曲线分析结果显示,血清SIGIRR、sTRAIL单独和二者联合预测IM患儿肝脏损伤的曲线下面积(area under the curve, AUC)分别为0.786、0.737、0.836,二者联合对IM患儿肝脏损伤预测效能的AUC大于SIGIRR、sTRAIL单独的预测效能。结论 IM患儿血清SIGIRR水平降低和sTRAIL水平升高与外周血T细胞亚群异常、肝脏损伤密切相关,血清SIGIRR、sTRAIL水平联合检测对IM患儿肝脏损伤的预测价值更高。

基于SIRT1/NRF2/HO-1信号通路探讨黄芪甲苷对大鼠肝星状细胞氧化损伤的作用机制

目的探讨黄芪甲苷治疗肝纤维化(HF)的作用机制。方法HSC-T6分为空白组、模型组、黄芪甲苷组、抑制剂组(采用Sirt1抑制剂EX527)、黄芪甲苷加抑制剂组,除空白组外均采用100μmol·L^(-1)的H_(2)O_(2)制造HSC-T6氧化应激的模型,模型组干预4 h,除模型组外干预24 h。ELISA法测定细胞上清中α-SMA、Collagen1水平;生化试剂盒法测定细胞中氧化应激相关指标;流式细胞术检测细胞中ROS的含量;免疫荧光法检测细胞中α-SMA的含量;实时荧光定量聚合酶链式反应(Real-time qPCR)和蛋白免疫印迹法(Western blot)检测各组细胞中Sirt1、Nrf2、HO-1、α-SMA、Collagen1 mRNA和蛋白表达水平。结果与空白组比较,经H_(2)O_(2)处理的HSC-T6上清中α-SMA、Collagen1及细胞中MDA、ROS的含量明显增加,而细胞中CAT含量及SOD活性明显降低,细胞中Sirt1、Nrf2、HO-1 mRNA和蛋白表达水平降低,α-SMA、Collagen1mRNA表达水平升高(P<0.05);与模型组比较,各给药组细胞中MDA、ROS的含量明显降低,CAT含量及SOD活性明显增加,黄芪甲苷组、黄芪甲苷加抑制剂组上清中α-SMA、Collagen1表达量降低,Sirt1、Nrf2、HO-1 mRNA和蛋白表达水平增加,α-SMA、Collagen1 mRNA表达水平降低(P<0.05)。结论黄芪甲苷可以减轻因H_(2)O_(2)刺激造成的HSC-T6氧化应激反应,减少氧化应激产物产生及胶原纤维沉积,从而达到抗HF的目的,其作用机制可能与调节Sirt1/Nrf2/HO-1信号通路有关。

Roles of homopolymeric apoferritin in alleviating alcohol-induced liver injury

Alcohol abuse induced liver damage are closely related to oxidative stress,several efforts have been made to quench free radicals produced in the liver.Heteropolymeric soybean seed ferritin has been proved to protect DNA from oxidative damage.However,the roles of homopolymeric apoferritin in quenching free radicals have been neglected so far.In order to improve the ferroxidase activity of ferritin,we have prepared apo ferritin by removing iron core from the cavity.In this study,the roles of apo shrimp ferritin(Marsupenaeus japonicus ferritin,MjFer)and human H chain ferritin(HuHF)in attenuating the hydroxyl radicals in vitro and in vivo has been investigated.Results showed that MjFer was able to quench the free radicals produced by iron oxidation more efficiently than HuHF in vitro.Animal studies using a mouse model of alcohol-induced acute liver injury revealed that MjFer is much more efficacious than HuHF in alleviating the liver injury,by attenuating oxidative stress and regulating Nrf2/HO1 expression.Accordingly,the higher activity of MjFer may be attributed to its faster iron oxidative rate than HuHF,which may help improve the intestinal integrity,and reduce abnormal iron absorption.As expected,iron contents in the liver of MjFer and HuHF group are much lower than that of model group,further alleviating the liver damage.These findings suggest that apoferritin may be a promising candidate for alcohol-induced liver injury.

连翘苷对酒精性肝损伤的保护作用

目的:研究连翘苷对酒精性肝损伤的保护作用。方法:人正常肝细胞株LO2于高糖DMEM完全培养基中培养,CCK-8试剂盒检测细胞存活率;ALT活性检测试剂盒检测细胞培养液中ALT的活性;DAPI染色法于荧光倒置显微镜下观察细胞核形态的改变;Western blotting检测凋亡相关蛋白PARP、caspase 3的表达。结果:连翘苷浓度依赖性地减轻酒精诱导的肝细胞损伤;DAPI染色结果表明连翘苷能够显著逆转酒精诱发的肝细胞核浓缩及核碎裂现象,细胞凋亡相关蛋白PARP和caspase 3的表达也被显著抑制。结论:连翘苷通过抑制肝细胞凋亡在酒精性肝损伤中发挥保护作用。

自体骨髓干细胞移植治疗慢性肝衰竭研究

目的探索自体骨髓干细胞移植对肝衰竭患者的治疗作用,为干细胞移植的临床应用研究提供基础。方法35例慢性重症肝病患者,20例行自体骨髓干细胞移植治疗,15例作为对照。在无菌条件下,从患者髂后上棘抽取骨髓30～50ml,分离纯化骨髓干细胞。在局部麻醉下行肝动脉介入,将分离的骨髓干细胞移植于肝脏。患者在移植后1、2、4、8周进行肝功能检测。观察患者移植后不同时间症状改善情况及术后不良反应情况。结果在移植8周后,患者丙氨酸转氨酸逐渐降低,由平均181.7μmolL降至72.1μmolL;总胆红素由平均153.8μmolL降至80.2μmolL;直接胆红素由平均74.1μmolL降至40.5μmolL;白蛋白逐渐升高,由平均26.5μmolL升至31.5μmolL;与对照组相比有明显差异,表明移植后患者肝功能明显改善。干细胞移植后凝血酶原活动度逐渐上升,由术前平均28.2%上升至50.1%。进一步观察自体骨髓干细胞移植对肝衰竭患者生存率的影响,发现移植后1周生存率为100%,4周为95%(1920),8周后为90%(1820),12周后为85%(1720),较对照组生存率明显升高。移植后大多数患者有明显症状改善,移植后8周内腹水减轻10例(50%),食欲改善15例(75%),体力好转11例(55%),腹胀减轻9例(45%)。在20例移植患者中未发现严重并发症,术后有轻度恶心1例,发热1例。结论自体骨髓干细胞移植治疗后,患者肝功能和凝血机制明显改善,生存率提高,症状好转,表明骨髓干细胞移植对肝衰竭患者治疗有效,安全,不良反应少。

双酚A对人胚肝细胞DNA损伤和修复功能的影响

[目的]研究双酚A(BisphenolA,BPA)对人肝L-02细胞DNA损伤修复功能的影响。[方法]分别以1、10、50、100μmol/LBPA和100μmol/LBPA+30μg/LVitC处理L-02细胞,比较处理和未处理的人胚肝L-02细胞在DNA损伤程度、切除修复鼠缺陷交叉互补蛋白1(ERCC1)、尿嘧啶DNA糖基化酶(UDG)、错配修复hMSH2基因(hMSH2)、DNA依赖蛋白激酶复合物(DNA-PKcs)及O6-甲基鸟嘌呤甲基转移酶(MGMT)表达水平的差异。每组细胞数均为1×106个/ml。[结果]BPA处理后,彗星试验显示BPA在低剂量(10μmol/L)时即具有DNA损伤作用(P<0.05),随着剂量增大,DNA损伤效应增加。100μmol/LBPA+30μg/LVitC组DNA损伤效应降低,显示抗氧化剂VitC可减缓BPA所致的DNA损伤效应,氧化损伤是BPA所致DNA损伤的方式之一。5种DNA损伤修复酶表达水平在1μmol/L、10μmol/L、50μmol/L剂量组依次降低,在100μmol/L、100μmol/L+VitC组依次增加,50μmol/L组各种DNA损伤修复酶表达水平最低。BPA50μmol/L组与溶剂对照比较,5种DNA损伤修复酶均有明显降低,差异具有显著性(P<0.05)。100μmol/L组的表达水平均高于50μmol/L组,100μmol/L+VitC组均高于100μmol/L组。除DNA-PKcs与hMSH2外,其他DNA损伤修复酶在100μmol/L+VitC组与溶剂对照组比较差异均无显著性(P>0.05)。[结论]BPA对L-02细胞具有氧化损伤作用,并可导致DNA损伤,多种DNA损伤修复酶参与修复双酚A所导致的DNA损伤。VitC可减缓双酚A的DNA损伤作用。

胚胎干细胞移植对急性损伤肝脏的作用

目的探索胚胎干细胞移植在急性损伤肝脏中的存活和分化情况。方法用四氯化碳(CCl4)制备小鼠急性肝损伤模型,将BrdU标记的来源于小鼠原始生殖细胞的胚胎干细胞经静脉移植于模型动物体内,分别于移植后2周、4周取出肝脏,用免疫组织化学、免疫荧光双重染色和PAS染色方法检测移植细胞在受体小鼠肝内的分布、存活、白蛋白的表达和糖原代谢。结果急性损伤小鼠肝脏可见大量肝细胞空泡样变性,有严重出血现象;2周时损伤肝内可见炎性细胞浸润和未吸收的出血区;4周时仍见少量炎性细胞浸润。干细胞移植2周,肝小叶内可见散在分布的BrdU和ALB双阳性着色细胞,PAS染色呈阳性反应;部分小细胞呈BrdU单阳性着色;损伤移植组的肝小叶结构基本恢复。移植4周,损伤肝小叶结构正常,小的BrdU阳性细胞数量增多。结论原始生殖细胞可以在急性损伤肝脏中存活,并分化为肝细胞和肝小叶内的其他细胞,并明显改善损伤肝的组织结构。

何首乌提取物对人正常肝细胞L02周期阻滞及凋亡的影响

目的:分析何首乌中致人肝细胞损伤物质的化学成分,初步探讨其致肝细胞损伤的机制。方法:生、制何首乌用70%乙醇提取,提取物经AB-8型大孔树脂吸附分离,依次用水、50%乙醇和95%乙醇洗脱得到生何首乌和制何首乌水洗脱物、50%乙醇洗脱物和95%乙醇洗脱物;生、制何首乌直接水煎煮得到水提物。体外培养人正常肝细胞L02,用含何首乌洗脱物和水提物的细胞培养液与细胞共同孵育一定时间后,检测药物对L02细胞生长的影响,筛选致肝细胞损伤成分。高效液相色谱法对有明显细胞毒作用的洗脱物进行成分分析,噻唑蓝法检测其对L02细胞增殖的影响,Giemsa染色进行形态学观察,流式细胞术检测L02细胞周期和凋亡情况。结果:生、制何首乌95%乙醇洗脱物对L02细胞有明显的生长抑制作用,其他组分对L02细胞的增殖无明显影响。成分分析显示,生、制何首乌95%乙醇洗脱物中大黄素含量最多,分别为(18.53±2.96)%和(10.28±1.34)%;何首乌95%乙醇洗脱物及大黄素可使L02细胞阻滞于S期,并诱导L02细胞凋亡。结论:何首乌95%乙醇洗脱物是何首乌中导致肝细胞损伤的主要物质,而大黄素是导致肝细胞损伤的成分之一。

大豆异黄酮对过氧化氢诱导的肝细胞损伤的保护作用

为探索大豆异黄酮对过氧化氢(H_2O_2)致肝细胞损伤的保护作用,以H_2O_2损伤Chang Liver细胞建立肝细胞氧化应激损伤模型,并以10,20和40 mg·L^(-1)大豆异黄酮进行干预。采用四甲基偶氮唑盐比色(MTT)法检测肝细胞存活率;以微板法检测细胞培养液中乳酸脱氢酶(LDH)、谷丙转氨酶(ALT)、谷草转氨酶(AST)活性和肝细胞超氧化物歧化酶(SOD)活性,以及肝细胞还原型谷胱甘肽(GSH)和丙二醛(MDA)含量;采用蛋白印迹技术检测肝细胞核因子-E2相关因子2(Nrf2)蛋白含量。在10~40 mg·L^(-1)范围内,大豆异黄酮对Chang Liver细胞不显示细胞毒作用。300μmol·L^(-1)H_2O_2刺激后,模型组肝细胞存活率与正常组比较下降,细胞外液中ALT、AST、LDH水平上升,说明Chang Liver细胞损伤显著;而模型组肝细胞中SOD和GSH水平与正常组比较下降,丙二醛水平上升,说明模型组Chang Liver细胞氧化应激增强。大豆异黄酮可剂量依赖性地提高Chang Liver细胞存活率,降低ALT、AST、LDH向细胞外液的释放;降低细胞MDA水平,升高细胞SOD和GSH水平,增高核中Nrf2蛋白水平。研究结果显示大豆异黄酮对H_2O_2致肝细胞氧化应激损伤具有保护作用。

溴虫腈对小鼠脾、肝、肾细胞DNA的损伤作用

[目的]探讨农药溴虫腈对小鼠脾、肝、肾细胞DNA是否有损伤作用及3种器官对溴虫腈的敏感性差异。[方法]溴虫腈按4.9、9.8、19.6mg/kg体重3个剂量,一次性灌胃染毒雄性小鼠,2 4h后用彗星试验检测3种脏器的彗星细胞率和彗星尾长。[结果]各剂量组的彗星细胞率和彗星尾长都显著增加(P <0 .0 5～P <0 .0 0 1) ;脾、肝、肾3种细胞的彗星细胞率及彗星尾长均具有剂量 反应或剂量 效应关系(前者r值依次为0 .9948,0 .9993 ,0 .9890 ,后者r值依次为0 .9999,0 .9999,0 .9898,P值均<0 .0 5 )。比较同剂量组不同器官之间的彗星细胞率,3个剂量组中均为肾细胞>肝细胞>脾细胞(P <0 .0 5～P <0 .0 1) ;比较同剂量组不同器官之间的彗星尾长,3个剂量组均为肾细胞>肝细胞>脾细胞(P<0 0 5 ) ,肝、脾细胞间差异无显著性。[结论]溴虫腈可诱发小鼠脾、肝、肾细胞DNA断裂损伤,肾细胞对溴虫腈致DNA断裂作用最为敏感。

污灌土壤有机提取物对原代大鼠肝细胞DNA损伤的研究

[目的]研究污灌土壤有机提取物对原代大鼠肝细胞DNA的损伤。[方法]选取A、B、C3类污灌农田土壤为研究对象,地下水灌溉的土壤D为对照,采用超声振荡法提取土壤中的有机污染物,并用彗星试验检测土壤有机提取物对原代大鼠肝细胞DNA的损伤情况。[结果]A、B、C、D3个剂量组(0.3g/ml,0.6g/ml,0.9g/ml)土壤有机提取物均可导致DNA迁移长度增加,拖尾率增高;A、B、C与D组相同染毒剂量比较,DNA迁移长度增加,拖尾率增高。[结论]A、B、C、D中均含有大量的DNA损伤物质,且A、B、C所含有机物的毒性要强于D.

自体骨髓干细胞移植治疗肝功能不全的临床研究

目的探索自体骨髓干细胞移植对肝功能不全患者的治疗效果,为干细胞移植的临床大规模开展提供基础。方法 6例肝功能不全患者均来源于我所住院患者,年龄44～69岁;患者在无菌条件下,从骼后上棘抽取骨髓50ml,分离纯化骨髓干细胞。在局部麻醉下,行肝动脉介入将分离的骨髓干细胞移植于肝脏;患者在移植后不同时间,1、2、4、8、12周进行肝功能检测;观察在移植后不同时间症状改善情况及术后的不良反应情况。结果移植后,谷丙转氨酶逐渐降低,由平均98.4IU/L降至41.5IU/L,总胆红素由平均136.5μmol/L降至78.4μmol/L,肌酐由112.3μmol/L降至72.1μmol/L;白蛋白逐渐升高,由平均23.3g/L升至32.6g/L;凝血酶原时间下降不明显。患者在移植12周生存率为100%,1例在7个月后死亡。移植后大多数患者症状明显改变,移植后12周食欲改善3例,精神体力好转3例,腹胀减轻2例。在4例随访患者中未发现严重并发症,术后有轻度恶心1例,发热1例,经对症治疗后好转,未见其他严重不良反应。结论自体骨髓干细胞移植治疗后,患者肝功能逐步改善,症状好转,表明骨髓干细胞移植对肝功能不全患者的治疗有效且副作用小,安全。

纳米TiO_2光诱导杀伤Bel-7402人体肝癌细胞研究

在光诱导条件下,采用HE染色法和四甲基偶氮唑蓝比色法(MTT法),研究了纳米TiO_2对Bel-7402人体肝癌细胞的杀伤作用,考察了在不同纳米TiO_2浓度、不同光照时间下纳米TiO_2的抑瘤效果,并且对抑瘤机制进行了探讨．结果发现：在光诱导条件下,适宜的了TiO_2浓度具有较高的抑瘤率,同时抑瘤过程表现出类似一级反应的动力学规律；在光诱导条件下,纳米TiO_2产生的活性氧组分与癌细胞膜内外的生物大分子反应,引起广泛的细胞结构破坏；造成癌细胞内Ca^(2+)离子稳态失去平衡；引发细胞微管相关蛋白2(MAP-2)表达的变化,促进微管发生重组,从而导致细胞凋亡和坏死．

HDV感染后在肝细胞损伤中的作用探讨

通过对ＨＤＶ 感染肝组织中ＨＤＡｇ 表达和细胞免疫状态的观察发现，ＨＤＡｇ 表达可在一定程度上直接引起肝细胞破坏，同时其表达数目与肝组织中炎症有关，特别是与汇管区中ＣＤ３ ＋ 细胞浸润（ｒ＝ ０－５２１） 和坏死区中ＣＤ８＋ 细胞浸润（ｒ＝ ０－４９５） 有较好相关性。提示ＨＤＶ 的直接细胞毒作用并非是引起肝细胞损伤的唯一因素，细胞免疫特别是ＣＤ８ ＋

当归多糖对小鼠酒精性肝细胞损伤的作用

目的探讨当归多糖对小鼠酒精性肝细胞损伤的保护机制。方法①通过一次性高浓度酒精灌胃建立小鼠急性酒精性肝损伤模型，之后将小鼠随机分为当归多糖组、酒精模型组，同时设正常对照组。当归多糖组连续腹腔注射当归多糖6d，酒精模型组及正常对照组腹腔注射等量生理盐水。动态观察小鼠的一般情况、肝组织形态学结构变化。②对各组血清丙二醛、超氧化物歧化酶及谷胱甘肽过氧化物酶进行生化检测。结果①小鼠酒精性肝损伤腹腔注射当归多糖组，肝组织逐渐得到明显修复，约11d肝小叶完全修复正常；而酒精性肝损伤未注射当归多糖组，肝组织修复差；正常对照组，肝组织结构无变化。②血清生化检测显示，比较当归多糖组与酒精模型组血清丙二醛、超氧化物歧化酶及谷胱甘肽过氧化物酶生化检测有统计学意义。结论当归多糖可通过提高抗氧化酶活性，清除氧自由基，减轻膜脂质过氧化链式反应，继而使肝细胞的生物膜结构恢复，从而对肝细胞起到保护作用。