本文選題：肝前體細(xì)胞（HPC） + 肝星狀細(xì)胞（HSC）�。� 參考：《第二軍醫(yī)大學(xué)》2014年碩士論文

【摘要】：【研究背景及目的】 肝前體細(xì)胞（Hepatic progenitor cell, HPC）是損傷肝臟中出現(xiàn)的一種具有雙向分化潛能的細(xì)胞，可向肝細(xì)胞和膽管上皮細(xì)胞分化參與肝再生。在慢性肝病狀態(tài)下，HPC可被激活參與肝損傷修復(fù)，而且HPC的數(shù)量與疾病的嚴(yán)重程度及肝癌發(fā)生的風(fēng)險指數(shù)相關(guān)。HPC既表達(dá)膽管上皮細(xì)胞標(biāo)志物（OV-6、PCK和Sox9等）和成熟肝細(xì)胞標(biāo)志物（白蛋白等），同時也表達(dá)肝母細(xì)胞標(biāo)志物（AFP和Dlk1等）。目前，HPC來源尚不明確，傳統(tǒng)觀點認(rèn)為HPC來自Hering管，也有研究顯示肝細(xì)胞、肝星狀細(xì)胞（Hepaticstellate cell, HSC）及骨髓干細(xì)胞在肝損傷環(huán)境下可轉(zhuǎn)化為HPC。因此，有學(xué)者認(rèn)為HPC可能是一種來源不同的異質(zhì)細(xì)胞群。 HSC位于Disse間隙，與肝細(xì)胞和肝竇內(nèi)皮細(xì)胞直接接觸。正常情況下，HSC處于靜止?fàn)顟B(tài)，肝損傷時，HSC可激活，產(chǎn)生細(xì)胞外基質(zhì)（Extracellular matrix, ECM），并分泌大量的細(xì)胞因子和生長因子。HSC的激活是肝纖維化發(fā)生發(fā)展的中心環(huán)節(jié)。近年來不斷有證據(jù)提示HSC還具有促進肝組織再生的作用，同時也有研究表明HPC可來源于HSC。這些研究均表明HSC在肝臟再生過程中扮演著重要角色。 基于以上背景，本課題旨在通過肝損傷動物模型明確活化HSC對HPC產(chǎn)生的影響，并利用體外共培養(yǎng)實驗進一步明確HPC的細(xì)胞來源。 【實驗方法】 一、探討HSC活化與HPC產(chǎn)生的關(guān)系 1、收集人急性肝炎、肝纖維化、肝硬化組織標(biāo)本，免疫熒光檢測HPC標(biāo)志物PCK及活化HSC標(biāo)志物α-SMA表達(dá)情況。 2、構(gòu)建2-乙酰氨基芴（2-acetylaminofluorene,2-AAF）/肝大部切除術(shù)（Partialhepatectomy, PH）動物模型誘導(dǎo)HPC活化。予2-AAF以10mg/kg體重的劑量對Sprague-Dawley（SD）大鼠進行灌胃，每天1次，持續(xù)5天，于第6天行肝大部切除術(shù)，手術(shù)當(dāng)日停止灌胃2-AAF，術(shù)后第2天繼續(xù)予2-AAF灌胃，并持續(xù)1周。分別取肝大部切除術(shù)后0天、4天、6天、9天的大鼠肝臟組織，免疫熒光檢測PCK及α-SMA表達(dá)情況。 3、構(gòu)建對乙酰氨基酚（N-acetyl-paraaminophen, APAP）肝損傷模型。按照300mg/kg體重的劑量對SD大鼠腹腔注射APAP，僅注射1次，注射24小時后處死大鼠，取新鮮肝臟組織，免疫熒光檢測PCK及α-SMA表達(dá)情況。 二、明確活化HSC對HPC產(chǎn)生的影響 （一）鑒定氯化釓（Gadolinium chloride, GdCl3）和膠霉毒素（Gliotoxin）對Kupffer細(xì)胞和活化HSC的作用 1、以2ml/kg體重的劑量對SD大鼠腹腔注射20%四氯化碳（Carbon tetrachloride,CCl4），隔日注射1次，共注射2次，末次注射后48小時，予GdCl3以10mg/kg體重的劑量尾靜脈注射1次。注射GdCl324小時后取大鼠肝臟組織。免疫熒光檢測Kuppfer細(xì)胞標(biāo)志物CD68和活化HSC標(biāo)志物α-SMA的表達(dá)情況。 2、對SD大鼠腹腔注射CCl4，隔日注射1次，共注射2次，在末次注射后48小時，予Gliotoxin以3mg/kg體重的劑量腹腔注射1次。注射Gliotoxin24小時后處死大鼠，取肝臟組織。免疫熒光檢測CD68和α-SMA的表達(dá)情況。 （二）觀察抑制HSC活化對HPC產(chǎn)生的影響 1、分離SD大鼠原代HSC，在體外予DMEM培液培養(yǎng)。收集體外培養(yǎng)后第3天（靜息）和第12天（活化）的HSC培養(yǎng)上清。 2、在2-AAF/PH模型肝大部切除術(shù)或APAP模型APAP注射前1天，予模型大鼠尾靜脈注射GdCl3抑制Kupffer細(xì)胞活性及HSC活化。分別于肝大部切除術(shù)或APAP注射72小時后處死大鼠，取肝臟組織，免疫熒光檢測PCK及α-SMA表達(dá)情況，對比觀察抑制HSC活化對HPC產(chǎn)生的影響。 3、在以上模型注射GdCl3抑制HSC活化后，于肝大部切除術(shù)或APAP注射的次日，腹腔注射不同活化狀態(tài)的原代HSC培養(yǎng)上清4ml1次。48小時后取大鼠肝臟組織，免疫熒光檢測PCK及α-SMA表達(dá)變化情況，對比觀察補充HSC培養(yǎng)上清對HPC產(chǎn)生的影響。 （三）觀察清除活化HSC對HPC產(chǎn)生的影響 1、在2-AAF/PH模型中，在灌胃2-AAF的第1、3天，同時以2ml/kg體重的劑量對SD大鼠腹腔注射20%CCl4以刺激HSC活化，并于肝大部切除術(shù)前1天，按3mg/kg體重的劑量對大鼠腹腔注射Gliotoxin清除活化HSC，于肝大部切除術(shù)后3天取大鼠肝臟組織，免疫熒光檢測PCK及α-SMA表達(dá)情況，比較清除活化HSC前后HPC產(chǎn)生數(shù)量的變化。 2、在2-AAF/PH+Gliotoxin處理的基礎(chǔ)上，于肝大部切除術(shù)后24小時腹腔注射不同活化狀態(tài)的原代HSC培養(yǎng)上清4ml1次。48小時后處死大鼠，免疫熒光檢測肝臟組織中PCK及α-SMA表達(dá)情況，明確活化HSC對HPC產(chǎn)生的影響。 三、明確活化HSC是否可促進大鼠肝細(xì)胞去分化為HPC 1、分離SD大鼠原代肝細(xì)胞和原代HSC，利用可使兩種細(xì)胞不直接接觸的雙層細(xì)胞共培養(yǎng)體系，將肝細(xì)胞分別與不同活化狀態(tài)HSC共培養(yǎng)，光鏡下觀察肝細(xì)胞的形態(tài)變化。在共培養(yǎng)的第0、3、5天，細(xì)胞免疫熒光檢測肝細(xì)胞標(biāo)志物HNF4α及HPC標(biāo)志物PCK、Sox9表達(dá)變化情況。2、為進一步明確與活化HSC共培養(yǎng)的肝細(xì)胞中出現(xiàn)的PCK陽性細(xì)胞具有雙向分化潛能，將其置于膽管上皮細(xì)胞分化誘導(dǎo)液（含丁酸鈉）中培養(yǎng)，8天后細(xì)胞免疫熒光檢測膽管上皮細(xì)胞標(biāo)志物CK19表達(dá)情況；將其置于肝細(xì)胞分化誘導(dǎo)液（含HGF、胰島素、轉(zhuǎn)鐵蛋白、地塞米松）中培養(yǎng)，11天后細(xì)胞免疫熒光檢測肝細(xì)胞標(biāo)志物白蛋白（Albumin, Alb）表達(dá)情況。 【實驗結(jié)果】 一、活化HSC促進HPC產(chǎn)生 （一）HPC產(chǎn)生與HSC活化相關(guān) 1、免疫熒光檢測人急性肝炎、肝纖維化、肝硬化組織中PCK及α-SMA的表達(dá)情況，結(jié)果顯示僅在與α-SMA陽性細(xì)胞毗鄰區(qū)域存在PCK陽性細(xì)胞。 2、在2-AAF/PH模型中，于肝大部切除術(shù)后0天、4天、6天、9天取肝臟組織，，免疫熒光結(jié)果顯示第0天和第4天時PCK陽性細(xì)胞及α-SMA陽性細(xì)胞數(shù)量均較少，但第6天和第9天時PCK陽性細(xì)胞及α-SMA陽性細(xì)胞數(shù)量明顯增加，且兩者緊密相聯(lián)，交織存在。 3、取APAP注射24小時后肝臟組織，免疫熒光結(jié)果顯示PCK陽性細(xì)胞被大量α-SMA陽性細(xì)胞緊密圍繞，提示HPC產(chǎn)生與HSC活化相關(guān)。 （二）活化HSC促進HPC產(chǎn)生 1、GdCl3可抑制Kuppfer細(xì)胞活性， Gliotoxin可清除活化HSC （1）CCl4造成急性肝損傷過程中，尾靜脈注射GdCl3，取肝組織，免疫熒光結(jié)果顯示與單純CCl4急性肝損傷標(biāo)本相比，CD68及α-SMA表達(dá)均明顯減少。 （2）在CCl4造成急性肝損傷后，腹腔注射Gliotoxin，免疫熒光檢測肝組織中α-SMA及CD68表達(dá)情況，結(jié)果發(fā)現(xiàn)與單純CCl4處理組相比，α-SMA陽性細(xì)胞明顯減少，而CD68陽性細(xì)胞卻沒有明顯變化。 2、抑制HSC活化對HPC產(chǎn)生的影響 （1）在2-AAF/PH及APAP造模過程中，利用GdCl3抑制Kuppfer細(xì)胞活性及HSC活化，免疫熒光結(jié)果顯示抑制HSC活化后，PCK陽性細(xì)胞及α-SMA陽性細(xì)胞數(shù)量均明顯減少。 （2）抑制HSC活化后，再補充不同活化狀態(tài)HSC培養(yǎng)上清，免疫熒光結(jié)果顯示補充活化HSC培養(yǎng)上清后，PCK陽性細(xì)胞及α-SMA陽性細(xì)胞數(shù)量增加，且兩者交織存在。而補充靜息HSC培養(yǎng)上清后，肝組織PCK及α-SMA表達(dá)沒有明顯變化。 3、清除活化HSC對HPC產(chǎn)生的影響 （1）與單純2-AAF/PH模型組織中存在大量PCK陽性細(xì)胞和α-SMA陽性細(xì)胞不同，腹腔注射Gliotoxin清除活化HSC后，免疫熒光結(jié)果顯示肝組織中僅存在極少量PCK陽性細(xì)胞和α-SMA陽性細(xì)胞。 （2）清除活化HSC后，再補充不同活化狀態(tài)HSC培養(yǎng)上清，結(jié)果顯示補充活化HSC培養(yǎng)上清后，PCK陽性細(xì)胞數(shù)量增加。而補充靜息HSC培養(yǎng)上清后，PCK陽性細(xì)胞數(shù)量卻沒有明顯變化。 二、活化HSC促進大鼠肝細(xì)胞去分化為HPC 1、將肝細(xì)胞與不同活化狀態(tài)HSC共培養(yǎng)，細(xì)胞免疫熒光結(jié)果顯示共培養(yǎng)5天時，與活化HSC共培養(yǎng)的肝細(xì)胞中可出現(xiàn)PCK陽性細(xì)胞，而與靜息HSC共培養(yǎng)或單獨培養(yǎng)的肝細(xì)胞中并沒有PCK陽性細(xì)胞的出現(xiàn)。在肝細(xì)胞與活化HSC共培養(yǎng)的第0、3、5天細(xì)胞免疫熒光檢測Sox9及HNF4α的表達(dá)情況，結(jié)果顯示第0天時Sox9表達(dá)極弱，而HNF4α大量表達(dá)，至第5天時，Sox9表達(dá)量明顯增多，而HNF4α表達(dá)減弱。 2、將PCK陽性細(xì)胞置于膽管上皮細(xì)胞分化誘導(dǎo)液中培養(yǎng)8天后，細(xì)胞免疫熒光顯示PCK陽性細(xì)胞分化為CK19陽性細(xì)胞。將PCK陽性細(xì)胞置于肝細(xì)胞分化誘導(dǎo)液中培養(yǎng)11天后，細(xì)胞免疫熒光結(jié)果顯示PCK陽性細(xì)胞中出現(xiàn)Alb陽性細(xì)胞。 【結(jié)論】 一、肝前體細(xì)胞的產(chǎn)生伴隨肝星狀細(xì)胞的活化。 二、活化肝星狀細(xì)胞促進肝前體細(xì)胞的產(chǎn)生。 三、活化肝星狀細(xì)胞促進大鼠肝細(xì)胞去分化為肝前體細(xì)胞。 四、肝細(xì)胞來源的肝前體細(xì)胞可向肝細(xì)胞和膽管上皮細(xì)胞分化。
[Abstract]:Background and purpose of research

In chronic liver disease , HPC can be activated to participate in liver injury repair , and the number of HPC is associated with the severity of the disease and the risk index of liver cancer . HPC is not yet clear . Traditional views suggest that HPC comes from Hering tube , and it also shows that the liver cells , hepatic stellate cells ( HSC ) and bone marrow stem cells can be transformed into HPC in the environment of liver injury . Therefore , it is thought that HPC may be a heterogeneous population of different sources .

HSC is located in the Disse Gap and is in direct contact with liver cells and endothelial cells . In normal circumstances , HSC can be activated to produce extracellular matrix ( ECM ) and secrete a large amount of cytokines and growth factors . The activation of HSC is a central link in the development of liver fibrosis . In recent years , it has been shown that HSCs also have the role of promoting the regeneration of liver tissue . These studies have shown that HSCs play an important role in liver regeneration .

Based on the above background , the aim of this study was to clarify the effect of activated HSC on HPC in animal model of liver injury , and to use in vitro co - culture experiment to further clarify the cell source of HPC .

EXPERIMENTAL METHOD

I . To explore the relationship between HSC activation and HPC production

1 . Collect human acute hepatitis , liver fibrosis , liver cirrhosis tissue specimen , immunofluorescence assay HPC marker PCK and activated HSC marker 偽 - SMA expression .

2 - Acetamidofluorene ( 2 - AAF ) / partial hepatectomy ( PH ) was used to induce HPC activation . 2 - AAF was administered to Sprague - Dawley ( SD ) rats at a dose of 10 mg / kg for 5 days .

3 . The model of hepatic injury of acetaminophen ( N - acetyl - parainfluenza , APAP ) was established . APAP was injected intraperitoneally with a dose of 300 mg / kg body weight . The rats were sacrificed only once . After 24 hours of injection , the rats were sacrificed , fresh liver tissue and immunofluorescence were used to detect PCK and 偽 - SMA expression .

II . Identify the effects of activated HSC on HPC

( 1 ) To identify the effects of gadolinium chloride ( GdCl3 ) and Gliotoxin on kupffer cells and activated HSC

1 . The rats were injected intraperitoneally with 20 % carbon tetrachloride ( CCl _ 4 ) at a dose of 2 ml / kg body weight for 2 times . After the last injection for 48 hours , GdCl3 was injected intravenously once a dose of 10 mg / kg body weight . The rat liver tissues were taken after the injection of GdCl324 hours . The expression of CD68 and activated HSC marker 偽 - SMA was detected by immunofluorescence .

The rats were sacrificed at 3 mg / kg body weight at 3 mg / kg after injection of Gliotoxin for 24 hours . The expression of CD68 and 偽 - SMA was detected by immunofluorescence assay .

( 2 ) To observe the effect of inhibiting HSC activation on HPC production

1 . The primary HSC of SD rats was isolated and cultured in vitro in DMEM culture . The culture supernatant was cultured on days 3 ( resting ) and 12 days ( activated ) after in vitro culture .

2 . After 2 - AAF / PH model hepatectomy or APAP model APAP injection , GdCl3 was injected into the tail vein of model rats to inhibit the activity of kupffer cells and the activation of HSC . After 72 hours of liver resection or APAP injection , the rats were sacrificed , liver tissues were sacrificed , PCK and 偽 - SMA expression were detected by immunofluorescence , and the effects of HSC activation on HPC were observed .

3 . After injection of GdCl3 by the above model , HSC activation was inhibited after HSC activation . After 48 hours , the changes of the expression of PCK and 偽 - SMA were detected in rat liver tissues and immunofluorescence assay , and the effect of supernatant on HPC was observed .

( iii ) Observe the effect of removing activated HSC on HPC

1 . In the 2 - AAF / PH model , the rats were injected intraperitoneally with 20 % carbon tetrachloride at 2 ml / kg for 1 day at the same time with 2 ml / kg body weight to stimulate the activation of HSC , and the rats were injected intraperitoneally with Gliotoxin for 1 day before hepatectomy .

2 . On the basis of 2 - AAF / PH + Gliotoxin treatment , the expression of PCK and 偽 - SMA in liver tissues was detected after 4 ml of HSC culture at 24 hours after hepatectomy , and the effects of activated HSC on HPC were clearly defined .

III . It is clear whether activated HSC can promote rat liver cells to differentiate into HPC .

1 . The primary hepatocytes and the primary HSC were isolated from SD rats . The hepatocytes were co - cultured with HSC of different activation states , and the morphological changes of hepatocytes were observed under light microscope . The expression of the hepatocyte marker HNF4 偽 and HPC markers PCK and Sox9 were observed under light microscope .
It was cultured in hepatocyte differentiation inducing solution ( including HGF , insulin , transferrin , dexamethasone ) , and the expression of liver cell marker albumin ( Albumin , albumin ) was detected after 11 days .

Results of experiment

I . Activate HSC to promote HPC production

( i ) HPC production related to HSC activation

1 . The expression of PCK and 偽 - SMA in acute hepatitis , liver fibrosis and liver cirrhosis were detected by immunofluorescence , and the results showed that PCK - positive cells were only in the area adjacent to 偽 - SMA positive cells .

2 . In the 2 - AAF / PH model , the number of PCK - positive cells and 偽 - SMA - positive cells were significantly increased at 0 days , 4 days , 6 days and 9 days after hepatectomy , but the number of PCK - positive cells and 偽 - SMA - positive cells increased significantly on the 6th and 4th days , and the number of PCK - positive cells and 偽 - SMA - positive cells increased significantly at the 6th and 9th days , and both were closely related and interleaved .

3 . After 24 - hour injection of APAP , the results showed that PCK - positive cells were closely surrounded by a large number of 偽 - SMA positive cells , suggesting that HPC production was related to HSC activation .

( 2 ) Activated HSC promotes HPC production

1 . GdCl3 can inhibit the activity of kuppfer cells . Gliotoxin can remove activated HSC .

( 1 ) In the course of acute liver injury , the expression of CD68 and 偽 - SMA was significantly decreased compared with the acute liver injury .

( 2 ) The expression of 偽 - SMA and CD68 in liver tissues was detected by intraperitoneal injection of Gliotoxin after the acute liver injury .

2 . Inhibition of HSC Activation on HPC Production

( 1 ) In the process of 2 - AAF / PH and APAP modeling , the cell activity and HSC activation were inhibited by GdCl3 . The results showed that the number of PCK - positive cells and 偽 - SMA positive cells decreased significantly after HSC activation .

( 2 ) After inhibiting HSC activation , HSC culture supernatants were supplemented with different activation states , and the results showed that the number of PCK - positive cells and 偽 - SMA positive cells increased and the number of PCK - positive cells and 偽 - SMA positive cells increased , and the expression of PCK and 偽 - SMA in liver tissue was not significantly changed after the culture supernatant of HSC .

3 . Effect of Removal of Activated HSC on HPC

( 1 ) There was a large number of PCK positive cells and 偽 - SMA positive cells in the tissues of the pure 2 - AAF / PH model , but only a small amount of PCK positive cells and 偽 - SMA positive cells were found in the liver tissues after intraperitoneal injection of Gliotoxin .

( 2 ) After removing activated HSCs , HSC culture supernatants were supplemented with different activation states . The results showed that the number of PCK - positive cells increased after the culture supernatant was supplemented with activated HSC , while the number of PCK - positive cells did not change significantly after the culture supernatant was supplemented .

2 . Activation of HSC promotes rat hepatocytes to differentiate into HPC

The expression of Sox9 and HNF4 偽 in hepatocytes co - cultured with activated HSC showed that Sox9 expression was very weak at day 0 , while the expression of HNF4 偽 increased significantly , while the expression of HNF4 偽 decreased .

2 . The PCK - positive cells were cultured for 8 days in the differentiation - inducing solution of bile duct epithelial cells . The cells of PCK - positive cells were differentiated into CK19 - positive cells .

Conclusion

First , the production of hepatic progenitor cells is accompanied by activation of hepatic stellate cells .

secondly , activating the hepatic stellate cells to promote the generation of the liver precursor cells .

thirdly , activating hepatic stellate cells to promote the dedifferentiation of rat liver cells into liver precursor cells .

4 . The liver precursor cells derived from hepatocytes can differentiate into hepatocytes and bile duct epithelial cells .

【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R575

【共引文獻(xiàn)】

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3 范公忍;李樹玲;;骨髓干細(xì)胞移植治療肝硬化的研究進展[J];國際消化病雜志;2013年04期

4 李榮富;王志紅;;干細(xì)胞與消化系統(tǒng)疾病的治療[J];國際消化病雜志;2013年04期

5 袁磊;羅賢武;王義;;肝內(nèi)膽管細(xì)胞癌發(fā)病機制研究進展[J];肝膽外科雜志;2013年05期

6 王波;唐曉鵬;;臍血干細(xì)胞與骨髓干細(xì)胞聯(lián)合移植治療大鼠急性肝衰竭實驗研究[J];實用肝臟病雜志;2014年01期

7 曾淑蘭;安運鋒;盧超;張立杰;張國海;;大鼠原代肝星形細(xì)胞的分離與鑒定[J];廣西師范大學(xué)學(xué)報(自然科學(xué)版);2014年01期

8 吳志偉;段文彪;劉贊;宋才鑫;段建文;俞富祥;張啟瑜;;肝星狀細(xì)胞對大鼠肝部分切除術(shù)后肝再生修復(fù)的影響[J];肝膽胰外科雜志;2014年03期

9 張玉霞;崔淑芳;張友磊;;乙酰肝素酶siRNA對部分肝切除大鼠術(shù)后肝再生的抑制作用[J];肝膽外科雜志;2014年04期

10 劉碩;黃宇;黃迪;翁杰鋒;張帥;古維立;;高齡供體肝移植安全性的探討[J];廣州醫(yī)藥;2014年06期

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5 閻麗;外周血單核細(xì)胞移植治療肝硬化的實驗研究[D];第四軍醫(yī)大學(xué);2008年

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7 羅宏武;人羊膜上皮細(xì)胞橫向分化為肝細(xì)胞樣細(xì)胞及脾內(nèi)移植的初步研究[D];中南大學(xué);2008年

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