虫草多糖的分离提纯及对体外肾细胞损伤和急性肾衰动物模型的保护与治疗作用

Separation and Purification of Cordyceps sinensis Polysaccharides and Their Protective and Therapeutic Effects on Renal Cell Injury in Vitro and Acute Renal Failure Animal Model

目的:探讨从虫草头孢菌粉中提取活性多糖组分,考察其对体外顺铂诱导大鼠肾细胞损伤和肌注甘油诱导急性肾衰大鼠模型的保护与治疗作用。方法:采用水提醇沉法从虫草头孢菌粉中提取分离多糖组分,同时通过MTT实验体外考察各组分对顺铂诱导MDCK和Vero细胞损伤的保护作用;根据体外活性筛选结果,选定活性虫草多糖组分,用于动物模型实验灌胃给药,进一步考察其对肌注甘油诱导急性肾衰大鼠模型的治疗作用,考察指标包括血液和尿液生化指标、24 h尿量、肾脏指数以及肾脏组织切片病理学检查。结果:从虫草头孢菌粉中提取分离得到组分A～H;经MTT实验体外筛选出其中活性较好的虫草多糖组分G,其灌胃给予肌注甘油诱导急性肾衰模型大鼠后,可显著降低模型大鼠血清总蛋白、尿素氮、肌酐和K+水平以及尿蛋白、尿酮体、尿红细胞和白细胞水平与肾脏指数,显著提高模型大鼠血清Na+水平和24 h尿量,并使模型大鼠肾脏病理学损伤明显改善。结论:采用水提醇沉法可从虫草头孢菌粉中有效分离提纯活性多糖组分,其中65%乙醇醇沉得到的虫草多糖组分为虫草中具肾脏保护作用的活性部位,对急性肾损伤有防治作用。

[Abstract] Objective：To explore the extraction of active polysaccharides from Cordyceps Cephalosporium powder and to investigate their protective and therapeutic effects on c/s-diaminodichloroplatin（ CDDP）- induced rat renal cell injury in vitro and intramuscularly injected glycerol-induced acute renal failure （ARF） rat model. Methods： The polysaccharide fractions were separated from Cordyceps Cephalosporium powder by water extraction and ethanol precipitation and their protective effects on CDDP-induced MDCK and Veto cell injury in vitro were evaluated using a MTY assay. Based on the in vitro screening result from the MTY assay, the active fraction was selected for using in an intramuscularly injected glycerol-induced ARF rat model experiment, in which its therapeutic effects by gastric perfusion on the rat model was further evaluated and the evaluation indicators included serum and urine biochemical parameters, 24 h-urinary production（UPD）, renal index and renal biopsy. Results： Fractions A-H were obtained from Cordyceps Cepha- lasporium powder, among which fraction G showed better activity in the MTT assay. After administration of fraction G to the intramuscularly injected glycerol-induced ARF rat model by gastric perfusion, the levels of serum total protein, urea nitrogen, creatinine and K ~ and the levels of urine protein, acetone body, red and white blood cells and renal index were significantly decreased, the level of serum Na ＋ and 24h-UPD were significantly increased and the renal pathological lesions were significantly improved in the ARF rat model. Conclusion： The active polysaccharides could be effectively extracted from Cordyceps Cephalosporium powder by water extraction and ethanol precipitation and the Cordyceps sinensis polysaccharides obtained by 65% ethanol precipitation are the active fraction having a renal protective effect in Cordyceps sinensis and potential to prevent and treat acute renal injury.