【摘要】：目的評估妊娠大鼠來源脂肪干細(xì)胞(adipose-derived stem cells,ADSCs)在急性肝損傷中的修復(fù)作用。方法取18周齡妊娠SD大鼠分離培養(yǎng)ADSCs并鑒定。另取SD大鼠20只隨機分為A、B、C、D 4組(n=5),A組大鼠不作處理,作為正常對照;B~D組大鼠腹腔注射CCl4制備急性肝損傷模型。造模后2 h,A組于大鼠脾臟注射DPBS,B組不注射任何試劑,C、D組分別于大鼠脾臟注射0.1 mL正常大鼠來源和妊娠大鼠來源ADSCs(2×10~6個/mL)。注射后7 d處死動物,取眼球血檢測血清中總膽紅素(total bilirubin,TBIL)、丙氨酸氨基轉(zhuǎn)移酶(alanine aminotransferase,ALT)、天門冬氨酸氨基轉(zhuǎn)移酶(aspartic acid transaminase,AST)、白蛋白(albumin,ALB)和肝功能總蛋白(total protein,TP)含量;取肝組織切片行HE染色檢測損傷肝組織形態(tài)學(xué)變化;免疫組織化學(xué)染色檢測肝組織中Ki67、甲胎蛋白(alpha-fetoprotein,AFP)和ALB的表達情況。結(jié)果 B組血清中TBIL、ALT、AST含量顯著高于A、C、D組,ALB、TP含量顯著低于A、C、D組,差異均有統(tǒng)計學(xué)意義(P0.05)。C、D組血清中TBIL、ALT、AST含量顯著高于A組,C組高于D組,差異均有統(tǒng)計學(xué)意義(P0.05);A、C、D組血清中ALB含量差異無統(tǒng)計學(xué)意義(P0.05);C、D組血清中TP含量顯著低于A組(P0.05),C、D組間比較差異無統(tǒng)計學(xué)意義(P0.05)。HE染色示:A組大鼠肝組織結(jié)構(gòu)清晰,細(xì)胞排列整齊,大小均勻;B組大鼠肝細(xì)胞明顯出現(xiàn)水腫,細(xì)胞排列紊亂,可見肝小葉內(nèi)點狀壞死,組織出現(xiàn)彌漫性炎性細(xì)胞浸潤;C、D組大鼠肝組織炎癥和肝細(xì)胞壞死程度較B組明顯減輕,同時由線粒體和粗面內(nèi)質(zhì)網(wǎng)擴張造成的空泡數(shù)量降低,其中D組肝損傷改善程度較C組更顯著。免疫組織化學(xué)染色示:C、D組Ki67陽性細(xì)胞率顯著高于A、B組,B組高于A組,D組高于C組,差異均有統(tǒng)計學(xué)意義(P0.05)。A、B組AFP均無表達,而C、D組陽性表達,D組AFP陽性細(xì)胞率顯著高于C組(t=3.006,P=0.017)。C、D組ALB表達均顯著高于A、B組,D組高于C組,差異均有統(tǒng)計學(xué)意義(P0.05)。結(jié)論與正常大鼠來源ADSCs移植比較,妊娠大鼠來源ADSCs移植后,肝損傷大鼠肝功能改善程度更顯著。
[Abstract]:Objective to evaluate the repair effect of adipose stem cells (adipose-derived stem cells,ADSCs) from pregnant rats in acute liver injury. Methods ADSCs was isolated from 18 week pregnant SD rats and identified. In addition, 20 SD rats were randomly divided into two groups: group A (n = 5), A) and group B (n = 5), A), and group B (n = 20) were injected intraperitoneally with CCl4 to make acute liver injury model. 2 hours after modeling, group A did not inject any reagents into the spleen of rats injected with DPBS,B. Group C D injected 0.1 mL ADSCs from normal rats and pregnant rats from ADSCs (2 脳 106 / mL).), respectively. Seven days after injection, the animals were killed and their eyeball blood was taken to detect serum total bilirubin (total bilirubin,TBIL), alanine aminotransferase (alanine aminotransferase,ALT), aspartate aminotransferase (aspartic acid transaminase,AST), albumin (albumin,). ALB and total liver protein (total protein,TP) content; The morphological changes of injured liver tissue were detected by HE staining and the expression of Ki67, alpha fetoprotein (alpha-fetoprotein,AFP) and ALB in liver tissue were detected by immunohistochemical staining. Results the levels of serum TBIL,ALT,AST and ALB,TP in group B were significantly higher than those in group A (P 0.05), and the levels of ALB,TP in group C were significantly lower than those in group D (P0.05). The content of TBIL,ALT,AST in group C was significantly higher than that in group A (P 0.05). Group C was higher than group D, the difference was statistically significant (P0.05). There was no significant difference in serum ALB content in group A (P 0.05). The content of serum TP in group C D was significantly lower than that in group A (P0.05), but there was no significant difference between group C and D (P0.05). HE staining showed that the liver tissue structure of group A was clear, cells were arranged neatly, and the size was uniform; In group B, there were edema and disarrangement of hepatocytes, dotted necrosis and diffuse inflammatory cell infiltration in liver lobule. The degree of liver inflammation and hepatocyte necrosis in group C was significantly reduced than that in group B, and the number of vacuoles caused by dilatation of mitochondria and rough endoplasmic reticulum was decreased. The improvement of liver injury in group D was more significant than that in group C. Immunohistochemical staining showed that the rate of Ki67 positive cells in group C was significantly higher than that in group A, group B and group C, respectively (P 0.05). There was no expression of AFP in group C, but no expression of AFP in group C was found in group C (P0.05), but the expression of AFP in group C was significantly higher than that in group C (P0.05). The positive expression rate of AFP in group D was significantly higher than that in group C (t 3.006). The expression of ALB in group C was significantly higher than that in group A (P 0.05). The expression of ALB in group D was significantly higher than that in group C (P0.05). Conclusion compared with the normal rat ADSCs transplantation, the liver function of the gravid rats was improved more significantly after the transplantation of the ADSCs derived from the pregnant rats.
【作者單位】： 昆明醫(yī)科大學(xué)第四附屬醫(yī)院骨與創(chuàng)傷外科;昆明醫(yī)科大學(xué)第二附屬醫(yī)院急診醫(yī)學(xué)部;
【分類號】：R575

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