冬虫夏草防治环孢素A肾毒性的体内与体外实验研究

CORDYCETP SINENSIS AMELIORATES CYCLOSPORIN A-INDUCED NEPHROTOXICITY:IN VIVO AND IN VITRO STUDIES

目的 :观察人工虫草对环孢素A肾毒性大鼠肾损害的保护作用 ;比较不同人工虫草组份对体外培养肾小管上皮细胞增生及环孢素A所致小管上皮细胞细胞毒性的影响。 方法 :以大剂量环孢素A持续皮下注射建立环孢素A肾毒性模型 ,观察人工虫草对大鼠血生化指标 ,肾功能以及肾组织病理改变的影响。同时体外培养肾近端小管上皮细胞系 ,观察虫草组份对细胞3H TdR掺入量及细胞周期的影响。测定细胞培养上清LDH酶活力 ,观察虫草对体外环孢素A小管毒性的影响。 结果 :环孢素A皮下注射四周时大鼠表现为血清白蛋白下降 ,血清肌酐及尿素氮轻度升高 ,血清钠离子浓度明显升高 ,大鼠尿NAG酶 ,溶菌酶明显升高 ,肾肌酐清除率明显降低。肾组织病理表现为片状小管上皮细胞萎缩 ,空泡变性 ,未见明显的间质纤维化。人工虫草治疗后大鼠尿NAG酶 ,溶菌酶明显降低 ,肾肌酐清除率明显改善 ,但肾组织病理检查未见明确改善。体外实验发现人工虫草可以促进培养肾小管上皮细胞增生 ,以水提部分小分子量组份的作用最强。并可改善环孢素A对体外培养肾小管上皮细胞的细胞毒性作用。其对人肾小管上皮细胞内c met以及EGF受体mRNA表达无明显上调作用。 结论 :体内及体外实验均证实人工虫草可以拮抗环孢素A的肾毒性作用。人工虫草水提部分中中?

To investigate the in vivo therapertic effects of Cordyceps Sinensis in CsA-incuced nephrotoxicity in rats,and to evaluate the effects of different ingredients isolated from Cordyceps Sinensis in promoting cultured human tubular epithelial cells proliferation and preventing CsA cytotoxicity in vitro. METHODOLOGY In vivo studies,CsA nephrotoxicity was induced in rats by high dosage CsA subcutaneous injection(15 mg/kg·d).Cordyceps Sinensis was given by gavage dayly after CsA injection.Kidney function,biochemical parameters and histopathology were examined four weeks later.In vitro studies,ingredients of different molecular weight isolated from Cordyceps Sinensis were used.Human kidney proximal tubular cell(HKC)proliferation was dectected by 3H-TdR incorporation and cell cycle analysis.LDH level in culture supernatant was dectected.Intracellular EGF and c-met mRNA expression was quantitated by RT-PCR assay. RESULTS In vivo studies,at the fourth week of observation,rats injected with CsA showed significantly elevated serum creatitinine,blood urea nitrogen(BUN),sodium level and markedly decreased serum albumin and creatinine clearance rate.Urinary NAG and lysozyme excretion elevated significantly.Histopathology revealed pachy tubular vacuolization at the second week,and pachy tubular atrophy at the fourth week.In Cordyceps Sinensis treated group,urinary NAG and lysozyme excretion,creatinine clearance rate were ameliorated markedly at the end of the experiment.Among the different ingredients from Cordyceps Sinensis,low molecular weight water extractant was the most potent in promoting HKC proliferation,as well as in the prevention of CsA-induced HKC cytotoxicity in vitro.EGF and c-met mRNA expression were not increased in HKC cells stimulated with Cordyceps Sinensis. CONCLUSION Cordyceps Sinensis can ameliorate CsA-induced nephrotoxicity in rats.Low molecular weight water extractant is the most potent in promoting HKC proliferation in vitro.