肝刺激因子通过抑制肝脏星状细胞活化减轻肝脏纤维化的研究
发布时间：2018-07-31 14:21
【摘要】：背景和目的肝脏受到损伤后,肝实质细胞发生坏死或凋亡,诱发炎症反应,分泌多种细胞因子,进而刺激静息状态的肝星状细胞(hepatic Stellate Cell,HSC)发生活化,进一步转分化为肌成纤维细胞(myofibroblast,MFB)。肌成纤维细胞可以分泌大量包含胶原在内的细胞外基质,导致肝脏纤维化。由此可见,HSC活化是肝纤维化发生与发展中的核心事件。鉴于HSC在肝脏纤维化发生发展过程中的重要作用,研究其活化的分子机制将有助于逆转或治疗肝脏纤维化。肝刺激因子(hepatic Stimulator Substance,HSS)是一种能刺激肝细胞增殖的生物活性物质。近期报道显示,HSS可通过保护线粒体膜孔道,维持细胞内钙离子稳态抑制细胞凋亡,发挥肝细胞保护作用,因而被认为是肝细胞内重要的存活因子(survival factor)。HSS与肝纤维化的发生发展也有一定关系。在小鼠肝纤维化模型中,过表达HSS可显著地减轻纤维化症状,而敲减HSS可明显加重小鼠CCl4和胆总管结扎诱导的肝纤维化(本实验室未发表资料),但HSS在肝脏纤维化发病与演化过程中的机制依然没有清楚地阐明。鉴于有报道称,HSS在肝实质细胞和HSC内均存在表达,我们不禁要问,HSS能否通过抑制肝星状细胞活化来达到减轻抑制肝脏纤维化的目的?为此,本实验培养HSC,通过调节其细胞内HSS表达水平,研究HSS对HSC活化的影响,并探讨其对肝纤维化产生的影响。本实验旨在从全新视角揭示HSS与HSC活化的关系,为预防及治疗肝纤维化提供理论基础。方法1、细胞模型的建立:选择并培养具有活化表型的肝星状细胞(LX-2)细胞系,并通过稳定转染的方法构建HSS高表达(HSS-Tx)和HSS敲减(HSS-sh RNA)的LX-2细胞系。2、HSC活化分子标记物的鉴定:通过western blot的方法检测HSS-Tx和HSSsh RNA细胞中HSC活化分子标记物a-SMA和Ⅲ型胶原的表达,以研究HSS与HSC活化之间的关系。3、活化型HSC细胞特征的检测:HSC活化特征包括增殖加快、迁移增强和凋亡减少。通过流式细胞仪分析细胞周期、MTS实验测定细胞活力和免疫荧光检测Ki67的阳性细胞数,研究HSS-Tx和HSS-sh RNA细胞增殖能力的变化;通过Transwell和Xcelligence实验检测两组细胞迁移能力的改变;并检测caspase-3/7活性以分析两组细胞的凋亡情况。4、HSS影响HSC活化的分子通路的探究:分别利用磷酸化MAPK抗体芯片和磷酸化酪氨酸激酶抗体芯片检测两组细胞差异活化的关键蛋白质,以研究HSS影响HSC活化的分子通路。5、HSS影响HSC细胞迁移的机制研究:分别利用高内涵细胞成像分析技术和激光共聚焦扫描显微镜检测细胞微丝的分布,纤维状-肌动蛋白(F-actin)和球状-肌动蛋白(G-actin)的含量,以分析两组细胞微丝的变化及其与迁移的关系。胞浆钙离子与微丝装配及细胞迁移关系密切,利用钙离子示踪剂显示胞浆内钙离子含量,反映钙离子变化和细胞运动的关系。6、HSS影响HSC线粒体动态性及线粒体钙离子的研究:HSS定位于HSC的线粒体,为探讨HSS是否通过影响线粒体功能进而改变HSC细胞迁移能力,首先利用活细胞染料标记两组细胞内的线粒体,并用激光共聚焦扫描显微镜和配套软件分析细胞的线粒体形态,通过ATP含量测定及Seahorse实验检测线粒体功能的变化,并使用线粒体钙离子示踪剂测定两组细胞线粒体钙离子的分布情况。7、回复实验:为进一步确定上述实验获得的实验结果,利用RNAi的方法抑制HSS-Tx细胞内HSS的表达,重复5和6的实验,即测定细胞内微丝的含量及分布,线粒体形态的变化等。结果1、Western blot结果显示,HSS转染的LX-2细胞(HSS-Tx)中HSS表达明显高于vector细胞,而且HSS-sh RNA转染的LX-2细胞(HSS-sh RNA)中HSS表达显著低于vector细胞,且在传代过程中能维持HSS高表达或低表达状态,说明细胞模型构建成功,可用于后续实验。2、Western blot结果显示,HSS-Tx细胞中a-SMA(P0.05)和III型胶原(P0.01)的表达明显低于HSS-sh RNA细胞,说明HSS可抑制HSC的活化。3、MTS法测定细胞活力结果显示,HSS-Tx组细胞活力在传代后36 h开始弱于HSS-sh RNA细胞(P0.01),而且这种状况一直持续至72 h(P0.05)。流式细胞仪结果显示,HSS-Tx组G0/G1期细胞比例明显多于HSS-sh RNA细胞。与此类似,其Ki67阳性细胞数也明显少于HSS-sh RNA(P0.05)。Transwell实验结果显示,在相同时间内穿过小孔的HSS-Tx细胞数目明显少于HSS-sh RNA细胞,同样Xcelligence实验结果显示在接种后1 h至25 h,HSS-Tx细胞的迁移能力均明显弱于sh RNA细胞(P0.0001),以上结果提示转染HSS基因可以抑制LX-2细胞的增殖及迁移。4、磷酸化MAPK通路抗体芯片结果显示,高表达HSS与敲减HSS两组细胞中,包括ERK,P38和AKT等在内的18种信号分子的磷酸化水平未见明显差异,而71个磷酸化酪氨酸激酶抗体芯片结果显示HSS-sh RNA细胞中Blk,Fyn,TNK1,FAK和TXK等与微丝装配相关激酶的磷酸化水平明显高于HSS-Tx组,提示HSS可能通过抑制上述激酶的活性,从而限制微丝装配。5、利用荧光标记的鬼笔环肽来标记HSS-Tx和HSS-sh RNA细胞内F-actin,通过高内涵细胞成像系统和激光共聚焦扫描显微镜显示F-actin含量和分布。结果显示,HSS-Tx组细胞内微丝数目明显少于HSS-sh RNA组(P0.01),而且分布更为不规则。利用同样的方法标记细胞内G-actin,结果显示HSS-Tx组细胞内G-actin的含量显著高于HSS-sh RNA组,提示HSS抑制HSC细胞微丝的装配,减少应力纤维的生成。细胞免疫荧光实验显示HSS-Tx组细胞内胞浆内钙离子的含量明显少于HSS-sh RNA组(P0.001)。6、利用Mito-tracker特异性标记活细胞线粒体,通过激光共聚焦扫描显微镜成像和相关软件分析显示,与HSS-sh RNA细胞相比,HSS-Tx细胞内线粒体形态变得更短而且更圆(P0.0001),而且Western blot结果显示,与线粒体融合相关的MFN-2表达降低(P0.01),说明HSS抑制了HSC细胞线粒体融合。测定细胞内线粒体氧代谢(采用Seahorse仪器),结果显示,HSS-Tx细胞的基础代谢能力下降,ATP含量明显低于HSS-sh RNA细胞(P0.0001)。线粒体融合受到抑制,不仅使线粒体功能受到破坏,而且也影响到线粒体钙离子稳态,进而影响整个胞浆钙离子稳态。利用线粒体钙离子示踪剂显示HSS高/低表达细胞内线粒体内钙离子的变化,结果显示,HSS-Tx细胞线粒体内的钙离子浓度明显低于HSS-sh RNA细胞(P0.0001)。7、Western blot结果显示,HSS-Tx细胞转染HSS si RNA pool后,HSS表达明显下调。转染HSS si RNA pool 3 d后,激光共聚焦扫描显微镜成像结果显示微丝分布开始变得规则,微丝含量明显增加,这些改变持续至转染7 d,而且更为明显。同样转染3 d后,线粒体开始变长,转染7 d时,线粒体形态改变最为明显;与之对应,细胞内ATP含量增加。结论1.HSS可以抑制HSCs活化。2.HSS破坏HSCs的线粒体动态性,使ATP合成减少,线粒体内钙离子含量下降,进而降低胞浆内钙离子浓度,抑制微丝装配,使HSCs活化与迁移能力下降。
[Abstract]:After the injury of the background and objective liver, the necrosis or apoptosis of liver parenchyma cells, induced inflammatory reaction, secreting a variety of cytokines, and then stimulating the resting state of hepatic Stellate Cell (HSC) to live, and further convert into myofibroblast (myofibroblast, MFB). Myofibroblast can secrete a large number of bags. The extracellular matrix, including collagen, leads to liver fibrosis. This shows that HSC activation is a core event in the development and development of liver fibrosis. In view of the important role of HSC in the development of liver fibrosis, the study of its activation molecular mechanism will help to reverse or treat liver fibrosis. Hepatic Stimulator Substance, HSS) is a bioactive substance that stimulates the proliferation of hepatocytes. Recently, it has been reported that HSS can protect the mitochondrial membrane pore, maintain intracellular calcium ion homeostasis and inhibit cell apoptosis and play a protective role in liver cells. Therefore, it is considered to be an important survival factor (survival factor).HSS in liver cells and the occurrence of liver fibrosis. Development also has some relationship. In the mouse liver fibrosis model, overexpression of HSS can significantly reduce the symptoms of fibrosis, while knockout HSS can obviously aggravate the liver fibrosis induced by CCl4 and choledochal ligation in mice. However, the mechanism of HSS in the pathogenesis and evolution of liver fibrosis is still not clearly elucidated. It is reported that HSS is expressed in both liver parenchyma cells and HSC. We can not help asking whether HSS can reduce hepatic fibrosis by inhibiting the activation of hepatic stellate cells. To this end, this experiment is to cultivate HSC and to study the effect of HSS on the activation of HSC by regulating the level of HSS expression in the cells, and to explore its effect on liver fibrosis. This experiment aims to reveal the relationship between HSS and HSC activation from a new perspective, and to provide a theoretical basis for the prevention and treatment of liver fibrosis. Method 1, the establishment of cell model: selecting and cultivating the activated phenotype of hepatic stellate cells (LX-2) cell lines, and constructing HSS high expression (HSS-Tx) and HSS knockout (HSS-sh RNA) LX through the stable transfection method. Identification of -2 cell line.2, HSC activation molecular markers: the expression of HSC activated molecular markers a-SMA and type III collagen in HSS-Tx and HSSsh RNA cells was detected by Western blot method to investigate the relationship between HSS and HSC activation. The cell cycle was analyzed by flow cytometry. The cell viability and immunofluorescence of Ki67 positive cells were measured by MTS test. The proliferation ability of HSS-Tx and HSS-sh RNA cells was studied. The changes in the cell migration ability of the two groups were detected by Transwell and Xcelligence tests, and the activity of caspase-3/7 was detected to analyze the apoptosis of the two groups of cells. .4, HSS affecting the molecular pathway of HSC activation: using phosphorylated MAPK antibody chip and phosphorylated tyrosine kinase antibody chip to detect the key protein of two groups of cell differentiation activation respectively, in order to study the mechanism of HSS affecting the HSC activation molecular pathway.5, HSS affecting the migration of HSC cells: high intension cell imaging analysis respectively The distribution of cell microfilaments, fibrous actin (F-actin) and spherical actin (G-actin) were detected by technique and laser confocal scanning microscope to analyze the changes of cell microfilaments and their relationship with migration in two groups. Cytoplasmic calcium ions were closely related to microfilament assembly and cell migration, and intracellular calcium tracers were used to display the cytoplasm. Calcium ion content, reflecting the relationship of calcium ion change and cell movement.6, HSS affects the dynamics of HSC mitochondria and the study of mitochondrial calcium ion: HSS is located in the mitochondria of HSC, to explore whether HSS can change the migration ability of HSC cells by influencing mitochondrial function and then change the mitochondria in two groups of cells by using living cell dyes. The morphology of mitochondria in cells was analyzed by laser confocal scanning microscope (confocal scanning microscope) and the software was used to detect the changes of mitochondrial function through ATP content determination and Seahorse test. The distribution of calcium ion in the mitochondria of two groups of cells was measured by mitochondrial calcium tracer.7, and the experimental results were further determined to determine the experimental results obtained by the above experiments. The expression of HSS in HSS-Tx cells was inhibited by RNAi, and 5 and 6 experiments were repeated, that was to determine the content and distribution of intracellular microfilament and the change of mitochondria morphology. Results 1, Western blot showed that the HSS expression in LX-2 cells (HSS-Tx) transfected by HSS was significantly higher than that of vector cells. The expression was significantly lower than that of vector cells, and the high expression or low expression of HSS could be maintained during the passage process, indicating that the cell model was successfully constructed and can be used in the follow-up experimental.2. The results of Western blot showed that the expression of a-SMA (P0.05) and III type collagen (P0.01) in HSS-Tx cells was significantly lower than that of HSS-sh RNA cells. The results of cell viability showed that the cell vitality of HSS-Tx group began to be weaker than HSS-sh RNA cells (P0.01) at 36 h after passage, and this condition continued to 72 h (P0.05). The result of flow cytometry showed that the proportion of G0/G1 stage in HSS-Tx group was more than HSS-sh RNA cells. 05) the results of.Transwell experiment showed that the number of HSS-Tx cells passing through the pores in the same time was significantly less than that of HSS-sh RNA cells. The same results showed that the migration ability of HSS-Tx cells was significantly weaker than sh RNA cells (P0.0001) at 1 h to 25 h after the inoculation. The results suggested that the transfection of the HSS gene could inhibit the proliferation of the cells. And migrating.4, the phosphorylated MAPK pathway antibody chip results showed that the phosphorylation level of 18 signal molecules, including ERK, P38 and AKT, was not significantly different in the high expression HSS and the knockout HSS two groups, while the 71 phosphorylated tyrosine kinase antibody chips showed Blk, Fyn, TNK1, etc. and microfilament in HSS-sh RNA cells. The level of phosphorylation of the kinases is obviously higher than that of the HSS-Tx group, suggesting that HSS may inhibit the assembly of.5 by inhibiting the activity of the above kinase, and using the fluorescent labeled phimic cyclic peptide to mark the F-actin in HSS-Tx and HSS-sh RNA cells. The content and distribution of F-actin are displayed by the high intension cell imaging system and the laser confocal scanning microscope. The results showed that the number of intracellular microfilaments in the HSS-Tx group was significantly less than that of the HSS-sh RNA group (P0.01), and the distribution was more irregular. The same method was used to mark the intracellular G-actin. The results showed that the content of G-actin in the HSS-Tx group was significantly higher than that of the HSS-sh RNA group, suggesting that HSS inhibits the assembly of HSC fine cell microfilaments and reduces the formation of stress fibers. Cell immunity is reduced. The intracellular calcium content in HSS-Tx group was obviously less than that of group HSS-sh RNA (P0.001).6, and the mitochondria of living cells were marked by Mito-tracker specificity. The morphology of mitochondria in HSS-Tx cells became shorter than that of HSS-sh RNA cells. More circular (P0.0001), and Western blot results showed that the expression of MFN-2 associated with mitochondrial fusion decreased (P0.01), indicating that HSS inhibited mitochondrial fusion of HSC cells. Intracellular mitochondrial mitochondrial oxygen metabolism (Seahorse instrument) was measured. The results showed that the basal metabolic ability of HSS-Tx cells decreased and ATP content was significantly lower than HSS-sh RNA cells. Mitochondrial fusion is inhibited, which not only destroys mitochondrial function, but also affects the homeostasis of mitochondrial calcium ion, and then affects the whole plasma calcium homeostasis. The mitochondrial calcium ion changes in HSS high / low expression cells are displayed by the mitochondrial calcium tracer. The results show that the calcium ion concentration in the mitochondria of HSS-Tx cells Significantly lower than HSS-sh RNA cell (P0.0001).7, Western blot results showed that HSS-Tx cells transfected HSS Si RNA pool, HSS expression decreased obviously. Laser confocal scanning microscopy showed that microfilament distribution began to become regular, microfilament content increased significantly, these changes continued to transfection of 7, and more After transfection of 3 D, mitochondria began to grow, and the mitochondrial morphologic changes were most obvious when transfected to 7 d, and the content of ATP in the cells increased. Conclusion 1.HSS inhibited the mitochondrial dynamics of HSCs activated.2.HSS, reduced the synthesis of ATP, decreased the content of calcium ions in the mitochondria, and then reduced the intracellular calcium concentration. Inhibition of microfilament assembly reduces HSCs activation and migration ability.
【学位授予单位】：首都医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R575.2
本文编号：2155862
[Abstract]:After the injury of the background and objective liver, the necrosis or apoptosis of liver parenchyma cells, induced inflammatory reaction, secreting a variety of cytokines, and then stimulating the resting state of hepatic Stellate Cell (HSC) to live, and further convert into myofibroblast (myofibroblast, MFB). Myofibroblast can secrete a large number of bags. The extracellular matrix, including collagen, leads to liver fibrosis. This shows that HSC activation is a core event in the development and development of liver fibrosis. In view of the important role of HSC in the development of liver fibrosis, the study of its activation molecular mechanism will help to reverse or treat liver fibrosis. Hepatic Stimulator Substance, HSS) is a bioactive substance that stimulates the proliferation of hepatocytes. Recently, it has been reported that HSS can protect the mitochondrial membrane pore, maintain intracellular calcium ion homeostasis and inhibit cell apoptosis and play a protective role in liver cells. Therefore, it is considered to be an important survival factor (survival factor).HSS in liver cells and the occurrence of liver fibrosis. Development also has some relationship. In the mouse liver fibrosis model, overexpression of HSS can significantly reduce the symptoms of fibrosis, while knockout HSS can obviously aggravate the liver fibrosis induced by CCl4 and choledochal ligation in mice. However, the mechanism of HSS in the pathogenesis and evolution of liver fibrosis is still not clearly elucidated. It is reported that HSS is expressed in both liver parenchyma cells and HSC. We can not help asking whether HSS can reduce hepatic fibrosis by inhibiting the activation of hepatic stellate cells. To this end, this experiment is to cultivate HSC and to study the effect of HSS on the activation of HSC by regulating the level of HSS expression in the cells, and to explore its effect on liver fibrosis. This experiment aims to reveal the relationship between HSS and HSC activation from a new perspective, and to provide a theoretical basis for the prevention and treatment of liver fibrosis. Method 1, the establishment of cell model: selecting and cultivating the activated phenotype of hepatic stellate cells (LX-2) cell lines, and constructing HSS high expression (HSS-Tx) and HSS knockout (HSS-sh RNA) LX through the stable transfection method. Identification of -2 cell line.2, HSC activation molecular markers: the expression of HSC activated molecular markers a-SMA and type III collagen in HSS-Tx and HSSsh RNA cells was detected by Western blot method to investigate the relationship between HSS and HSC activation. The cell cycle was analyzed by flow cytometry. The cell viability and immunofluorescence of Ki67 positive cells were measured by MTS test. The proliferation ability of HSS-Tx and HSS-sh RNA cells was studied. The changes in the cell migration ability of the two groups were detected by Transwell and Xcelligence tests, and the activity of caspase-3/7 was detected to analyze the apoptosis of the two groups of cells. .4, HSS affecting the molecular pathway of HSC activation: using phosphorylated MAPK antibody chip and phosphorylated tyrosine kinase antibody chip to detect the key protein of two groups of cell differentiation activation respectively, in order to study the mechanism of HSS affecting the HSC activation molecular pathway.5, HSS affecting the migration of HSC cells: high intension cell imaging analysis respectively The distribution of cell microfilaments, fibrous actin (F-actin) and spherical actin (G-actin) were detected by technique and laser confocal scanning microscope to analyze the changes of cell microfilaments and their relationship with migration in two groups. Cytoplasmic calcium ions were closely related to microfilament assembly and cell migration, and intracellular calcium tracers were used to display the cytoplasm. Calcium ion content, reflecting the relationship of calcium ion change and cell movement.6, HSS affects the dynamics of HSC mitochondria and the study of mitochondrial calcium ion: HSS is located in the mitochondria of HSC, to explore whether HSS can change the migration ability of HSC cells by influencing mitochondrial function and then change the mitochondria in two groups of cells by using living cell dyes. The morphology of mitochondria in cells was analyzed by laser confocal scanning microscope (confocal scanning microscope) and the software was used to detect the changes of mitochondrial function through ATP content determination and Seahorse test. The distribution of calcium ion in the mitochondria of two groups of cells was measured by mitochondrial calcium tracer.7, and the experimental results were further determined to determine the experimental results obtained by the above experiments. The expression of HSS in HSS-Tx cells was inhibited by RNAi, and 5 and 6 experiments were repeated, that was to determine the content and distribution of intracellular microfilament and the change of mitochondria morphology. Results 1, Western blot showed that the HSS expression in LX-2 cells (HSS-Tx) transfected by HSS was significantly higher than that of vector cells. The expression was significantly lower than that of vector cells, and the high expression or low expression of HSS could be maintained during the passage process, indicating that the cell model was successfully constructed and can be used in the follow-up experimental.2. The results of Western blot showed that the expression of a-SMA (P0.05) and III type collagen (P0.01) in HSS-Tx cells was significantly lower than that of HSS-sh RNA cells. The results of cell viability showed that the cell vitality of HSS-Tx group began to be weaker than HSS-sh RNA cells (P0.01) at 36 h after passage, and this condition continued to 72 h (P0.05). The result of flow cytometry showed that the proportion of G0/G1 stage in HSS-Tx group was more than HSS-sh RNA cells. 05) the results of.Transwell experiment showed that the number of HSS-Tx cells passing through the pores in the same time was significantly less than that of HSS-sh RNA cells. The same results showed that the migration ability of HSS-Tx cells was significantly weaker than sh RNA cells (P0.0001) at 1 h to 25 h after the inoculation. The results suggested that the transfection of the HSS gene could inhibit the proliferation of the cells. And migrating.4, the phosphorylated MAPK pathway antibody chip results showed that the phosphorylation level of 18 signal molecules, including ERK, P38 and AKT, was not significantly different in the high expression HSS and the knockout HSS two groups, while the 71 phosphorylated tyrosine kinase antibody chips showed Blk, Fyn, TNK1, etc. and microfilament in HSS-sh RNA cells. The level of phosphorylation of the kinases is obviously higher than that of the HSS-Tx group, suggesting that HSS may inhibit the assembly of.5 by inhibiting the activity of the above kinase, and using the fluorescent labeled phimic cyclic peptide to mark the F-actin in HSS-Tx and HSS-sh RNA cells. The content and distribution of F-actin are displayed by the high intension cell imaging system and the laser confocal scanning microscope. The results showed that the number of intracellular microfilaments in the HSS-Tx group was significantly less than that of the HSS-sh RNA group (P0.01), and the distribution was more irregular. The same method was used to mark the intracellular G-actin. The results showed that the content of G-actin in the HSS-Tx group was significantly higher than that of the HSS-sh RNA group, suggesting that HSS inhibits the assembly of HSC fine cell microfilaments and reduces the formation of stress fibers. Cell immunity is reduced. The intracellular calcium content in HSS-Tx group was obviously less than that of group HSS-sh RNA (P0.001).6, and the mitochondria of living cells were marked by Mito-tracker specificity. The morphology of mitochondria in HSS-Tx cells became shorter than that of HSS-sh RNA cells. More circular (P0.0001), and Western blot results showed that the expression of MFN-2 associated with mitochondrial fusion decreased (P0.01), indicating that HSS inhibited mitochondrial fusion of HSC cells. Intracellular mitochondrial mitochondrial oxygen metabolism (Seahorse instrument) was measured. The results showed that the basal metabolic ability of HSS-Tx cells decreased and ATP content was significantly lower than HSS-sh RNA cells. Mitochondrial fusion is inhibited, which not only destroys mitochondrial function, but also affects the homeostasis of mitochondrial calcium ion, and then affects the whole plasma calcium homeostasis. The mitochondrial calcium ion changes in HSS high / low expression cells are displayed by the mitochondrial calcium tracer. The results show that the calcium ion concentration in the mitochondria of HSS-Tx cells Significantly lower than HSS-sh RNA cell (P0.0001).7, Western blot results showed that HSS-Tx cells transfected HSS Si RNA pool, HSS expression decreased obviously. Laser confocal scanning microscopy showed that microfilament distribution began to become regular, microfilament content increased significantly, these changes continued to transfection of 7, and more After transfection of 3 D, mitochondria began to grow, and the mitochondrial morphologic changes were most obvious when transfected to 7 d, and the content of ATP in the cells increased. Conclusion 1.HSS inhibited the mitochondrial dynamics of HSCs activated.2.HSS, reduced the synthesis of ATP, decreased the content of calcium ions in the mitochondria, and then reduced the intracellular calcium concentration. Inhibition of microfilament assembly reduces HSCs activation and migration ability.
【学位授予单位】：首都医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R575.2
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