人脐带间充质干细胞治疗环磷酰胺导致的大鼠药物性肝损伤

Human umbilical cord mesenchymal stem cells alleviate cyclophosphamide-induced liver injury in rats

目的研究脐带间充质干细胞(umbilical cord-mesenchyma stem cell,UC-MSC)对环磷酰胺(cyclophosphamide,CTX)导致的大鼠药物性肝损伤的治疗作用.方法首先分离培养UC-MSC.SD大鼠腹腔注射CTX建立肝损伤模型,分别给予尾静脉注射UC-MSC或生理盐水,将其分为Control组、C TX组和CTX+UC-MSC组.在不同时间点检测血清谷丙转氨酶(alanine transaminase,ALT)、谷草转氨酶(aspartate transaminase,AST)、碱性磷酸酶(alkaline phosphatase,ALP)和总胆红素(total bilirubin,TBIL)的浓度.肝组织均浆中检测丙二醛(malondialdehyde,MDA)、脂质过氧化物(lipid peroxide,LPO)、一氧化氮(NO)、总超氧化物歧化酶(total superoxide dismutase,T-SOD)、谷胱甘肽(glutathione,GSH)、谷胱甘肽过氧化物(glutathione peroxidase,GSH-PX)的水平.qPCR检测肝组织中Bax、Bcl-2、血管内皮细胞生长因子(vascular endothelial growth factor,V E G F)A基因m R N A表达情况.肝组织切片行H E染色和α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、Ki-67染色.结果与Control组相比,CTX组ALT、AST、ALP和TBIL含量明显增高,SOD、GSH和GSH-PX表达下降,MDA、LPO和NO表达增加,凋亡增加,VEGFA表达减少,肝细胞水肿明显.CTX+UC-MSC组ALT、AST、ALP和TBIL值均低于CTX组(P<0.05).CTX+UC-MSC组的SOD、GSH和GSH-PX均高于CTX组,MDA、LPO和NO均低于CTX组(P<0.05).CTX+UC-MSC组的Bax的表达低于CTX组,BCL-2和VEGFA的m RNA水平高于CTX组(P<0.05).HE染色示CTX+UC-MSC组的肝细胞水肿、出血等损伤低于CTX组.CTX+UC-MSC组的α-SMA^+细胞少于CTX组,Ki-67^+细胞多于CTX组(P<0.05).结论 UC-MSC治疗可缓解CTX导致的大鼠药物性肝损伤.

AIM To assess the effect of human umbilical cord mesenchymal stem cells（UC-MSCs） on cyclophosphamide（CTX）-induced liver injury.METHODS UC-MSCs were isolated from the human umbilical cord. Male SD rats were randomly divided into three groups： control group, CTX group, and CTX ＋ UCMSC group. The CTX group and CTX ＋ UC-MSC group were intraperitoneally injected with CTX. After that, the control group and CTX group were injected with water via the tail vein, and the CTX ＋ UCMSC group was injected with MSCs via the tail vein. Six rats of each group were selected randomly and sacrificed at different time points. Blood samples were taken to measure serum alanine transaminase（ALT）, aspartate transaminase（AST）, alkaline phosphatase（ALP）, and total bilirubin（TBIL）. Liver tissues were collected for biochemical assays of malondialdehyde（MDA）,lipidperoxide（LPO）,NO,glutathione（GSH）, glutathione peroxidase（GSH-PX）, and total superoxide dismutase（T-SOD）. q PCR was used to test the expression of Bcl-2, Bax, and vascular endothelial growth factor（VEGF） A. Histological examination（HE straining） and immunohistochemical staining for α-smooth muscle actin（α-SMA） and Ki-67 were also performed.RESULTS Compared with the control group, the levels of ALT, AST, ALP, and TBIL were significantly higher, the SOD, GSH, and GSH-PX contents were significantly lower,MDA,LPO,and NO were significantly higher, cell apoptosis significantly increased, and the expression of VEGFA significantly decreased in the CTX group. Compared with the CTX group, the ALT, AST, ALP, and TBIL were significantly lower, the SOD, GSH, and GSH-PX contents were significantly higher, and MDA, LPO, and NO were significantly lower in the CTX ＋ UC-MSC group（P 0.05）. The expression of Bax was lower and Bcl-2 and VEGFA expression was higher in the CTX ＋ UC-MSC group than in the CTX group（P 0.05）. Pathological analysis showed that liver status was better in the CTX ＋ UC-MSC group than in the CTX group. The α-SMA~＋ cells in the CTX ＋ UC-MSC group were less than and Ki-67~＋ cells were more than those in the CTX group（P 0.05）. CONCLUSION UC-MSCs injected via the tail vein could alleviate CTXinduced hepatotoxicity in rats.