于雅娟.苦瓜提取物联合脂肪干细胞移植对顺铂所致大鼠肾损伤的影响[J].转化医学杂志,2020,9(4):210-214
苦瓜提取物联合脂肪干细胞移植对顺铂所致大鼠肾损伤的影响
Effect of Momordica charantia L. extract combined with ADSCs transplantation on cisplatin-induced renal injury in rats
  
DOI:
中文关键词:  肾损伤  苦瓜提取物  脂肪干细胞  顺铂  大鼠
英文关键词:Kidney injury  Momordica charantia L. extract  Adipose-derived stem cells(ADSCs)  Cisplatin  Rat
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作者单位
于雅娟 赤峰学院附属医院肾内科 
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中文摘要:
      目的探讨苦瓜提取物联合脂肪干细胞移植对顺铂所致大鼠肾脏损伤的保护机制。方法复苏培养脂肪干细胞后行PKH-26标记。制备顺铂肾损伤大鼠模型,随机分为模型组、脂肪干细胞组、苦瓜提取物组、联合治疗组,15只/组,随机选取15只大鼠作为对照组。造模成功后24 h脂肪干细胞组大鼠经尾静脉注射0.5 mL 1×108/L脂肪干细胞悬液,苦瓜提取物组大鼠灌胃80 mg/kg苦瓜提取物,联合治疗组大鼠同时给予脂肪干细胞悬液和苦瓜提取物,对照组和模型组尾静脉注射0.5 mL DMEM/F12培养液,1/d,连续进7 d。7 d后检测大鼠血肌酐、血尿素氮水平和尿N-乙酰-β-D-葡萄(N-acetyl-b-D-glucosaminidase,NAG)、尿γ-谷氨酰转肽酶(γ-glutamyltranspeptidase,γ-GT)、尿蛋白含量,取大鼠肾组织行苏木精-伊红染色、TUNEL染色、RT-PCR及Western Blot检测。结果与对照组比较,模型组血肌酐、血尿素氮含量及尿NAG、γ-GT、尿蛋白升高(P<0.05),苦瓜提取物组、脂肪干细胞组以上指标含量较模型组降低(P<0.05),联合治疗组明显降低(P<0.05)。联合治疗组的病理改善情况优于脂肪干细胞组和苦瓜提取物组,脂肪干细胞组和苦瓜提取物组较模型组改善,但不及联合治疗组。PKH-26标记的脂肪干细胞,联合治疗组多于脂肪干细胞组,模型组及苦瓜提取物组未见PKH-26阳性细胞。TUNEL细胞凋亡数结果显示模型组最多,而对照组最少,且联合治疗组低于苦瓜提取物组和脂肪干细胞组(P<0.05)。与对照组比较,模型组RAGE、ROS、Bax的mRNA和蛋白表达水平上调(P<0.05),Bcl-2的mRNA和蛋白表达水平下调(P<0.05)。与模型组比较,苦瓜提取物组、脂肪干细胞组RAGE、ROS、Bax的mRNA和蛋白表达水平下调(P<0.05),Bcl-2 mRNA和蛋白表达水平上调(P<0.05)。联合治疗组RAGE、ROS、Bax的mRNA和蛋白表达水平下调较苦瓜提取物组、脂肪干细胞组明显(P<0.05),Bcl-2的mRNA和蛋白表达水平上调亦明显(P<0.05)。结论苦瓜提取物联合脂肪干细胞移植可能通过调控RAGE-ROS信号通路减少肾细胞凋亡发挥对顺铂致大鼠肾损伤的保护作用。
英文摘要:
      ObjectiveTo explore the protective effect and mechanism of Momordica charantia L. extract combined with adipose-derived stem cells(ADSCs) transplantation on cisplatin-induced rat kidney injury. MethodsADSCs were cultured in vitro and labeled with PKH-26. A rat model of cisplatin nephrotoxicity was prepared and randomly divided into model group, ADSc group, Momordica charantia L. extract group, and combined treatment group, 15 rats in each group, and another 15 rats were taken as normal control group. 0.5 mL DMEM was injected into tail vein of rats in pathological model group, ADSCs suspension was injected into tail vein of rats in ADSCs group, rats in Momordica charantia L. extract group were gavaged with Momordica charantia L. extract, and rats in combination group were given the above two treatments at the same time. After 7 days of treatment, the changes of serum creatinine and blood urea nitrogen, urinary N-acetyl-b-D-glucosaminidase (NAG), γ-glutamyltranspeptidase (γ-GT) and urinary protein were measured. The pathological changes of kidney tissues and the distribution of PKH-26 labeled cells were observed. The apoptosis of kidney cells was detected by TUNEL method. RAGE-ROS signaling pathway and the expression of Bax and Bcl-2 in kidney tissues were detected by RT-PCR and Western Blot. ResultsCompared with the normal control group, the levels of serum creatinine and blood urea nitrogen and urinary NAG, γ-GT and urine protein were increased (P<0.05), and the above indicators of Momordica charantia L. extract group and ADSCs group was lower than that of the pathological model group (P<0.05), and the combined group was significantly lower (P<0.05). Pathological results showed that the ADSCs group and the Momordica charantia L. extract group were improved compared with the pathological model group, but not as compared with the combined group. PKH-26-labeled ADSCs, the combined group was more than the ADSCs group, and the pathological model group and the Momordica charantia L. extract were not found. The results of TUNEL cell apoptosis showed that the single cisplatin pathological model group was the most, while the control group was the least and the combined group was known to be lower than the Momordica charantia L. extract group and the ADSCs group (P<0.05). The expressions of RAGE, ROS and Bax mRNA and protein were the lowest in the normal control group, the lower in the combined group, the higher in the Momordica charantia L. extract group and the ADSCs group, and the highest in the pathological model group (P<0.05). The expression of Bcl-2 mRNA and protein was the highest in the normal control group, and the combination group was higher, the Momordica charantia L. extract group and the ADSCs group were lower, and the pathological model group was the lowest (P<0.05). ConclusionMomordica charantia L. extract combined with ADSCs transplantation may play a protective role in cisplatin-induced renal injury by regulating RAGE-ROS signaling pathway and reducing renal cell apoptosis.
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