右归饮对肾虚雄性大鼠生殖功能损害的改善作用及其与EPO的相关性
|
摘要
目的:观察右归饮对腺嘌呤所致肾虚生殖功能损害大鼠的改善作用及与EPO的相关性。方法:将SD大鼠随机分为正常组15只与造模组85只,造模组用腺嘌呤150mg·kg-1·d-1连续灌胃14d。以检测血液肾功能、生殖激素作为判定模型成功的标准。将造模成功大鼠随机分为5组:模型组,龟鹿二仙胶10g/kg,右归饮10、20、40g/kg组,各15只,并进行组间均衡性检验。第15天开始,除正常组外,各组隔日给予150mg/kg腺嘌呤灌胃维持模型,同时右归饮各组与龟鹿二仙胶组灌胃给药每日1次。第46天麻醉取血,处死。检测各组大鼠血清中肌酐(CREA)、尿素(UREA)、腺体激素、生殖相关激素的含量;HE染色检测组织病理变化;免疫组化法检测及免疫印迹法检测肾脏与睾丸中EPO介导的HIF1α-STAT5通路中关键蛋白的含量。结果:与正常组比较,模型组大鼠血清中UREA、CREA含量显著上升,ACTH、T3、EPO含量显著下降,同时血清中E2、T、LH、DHT含量显著下降(P<0.05,P<0.01)。与模型组比较,右归饮各剂量均能不同程度地降低大鼠血清中UREA、CREA含量,增加血清中ACTH、T3、EPO含量(P<0.05,P<0.01);缓解大鼠肾脏及睾丸组织病理变化;均能显著调节肾脏与睾丸中EPO介导的HIF1α-STAT5通路中关键蛋白的表达。结论:EPO是腺嘌呤致大鼠肾虚生殖功能低下的重要相关因子,右归饮对腺嘌呤所致肾虚生殖功能低下大鼠异常指标均有显著的逆转作用,其作用机制与上调血清中EPO含量,继而调节肾脏和睾丸中EPO介导的HIF1α-STAT5通路中关键蛋白表达有关。
Objective: To observe the improvement effect of Yougui Drink on rats with adenture-induced kidney dysfunction and its correlation with EPO. Methods: SD rats were randomly divided into normal group(15) and model group(85),and the model was administered with adenine 150 mg·kg-1·d-1 for 14 days. To test blood kidney function and reproductive hormone as criteria for judging the success of the model. Rats with successful modeling were randomly divided into 5 groups:model group, Guilu Erxian cream 10 g/kg group, Yougui Drink 10 g/kg group, 20 g/kg group, 40 g/kg group, 15 in each group,and the balance test between groups were carried out. On the 15 th day, except for the normal group, each group was given a150 mg/kg adenine gavage maintenance model every other day, and the right group and the Guilu Erxian Cream group were intragastrically administered once a day. On the 46 th day, blood was anesthetized and sacrificed. Serum levels of creatinine, urea,glandular hormone and reproductive related hormones were detected in each group. HE staining was used to detect histopathological changes; Immunohistochemistry and immunoblotting were used to detect EPO in kidney and testis. Mediates the content of key proteins in the HIF1α-STAT5 pathway. Results: Compared with the normal group, the serum levels of creatinine and urea in the model group increased significantly, and the contents of ACTH, T3 and EPO decreased significantly(P<0.05, P<0.01). Compared with the model group, each dose group of Yougui Drink can reduce the serum creatinine and urea content in rats, increase the content of ACTH and EPO in serum(P<0.05, P<0.01), and relieve the pathological changes of rat kidney and testis. Yougui Drink can significantly regulate EPO-mediated key protein HIF1α in HIF1α-STAT5 pathway in kidney and testis expression. Conclusion: EPO is an important factor in the hypoplasia of kidney deficiency caused by adenine. The Yougui Drink has a significant reversal effect on the abnormal index of rats with hypothyroidism caused by adenine; the Yougui Drink and up-regulate the EPO content in serum, and then regulate the key protein expression of EPO-mediated HIF1α-STAT5 pathway in kidney and testis.
Objective: To observe the improvement effect of Yougui Drink on rats with adenture-induced kidney dysfunction and its correlation with EPO. Methods: SD rats were randomly divided into normal group(15) and model group(85),and the model was administered with adenine 150 mg·kg-1·d-1 for 14 days. To test blood kidney function and reproductive hormone as criteria for judging the success of the model. Rats with successful modeling were randomly divided into 5 groups:model group, Guilu Erxian cream 10 g/kg group, Yougui Drink 10 g/kg group, 20 g/kg group, 40 g/kg group, 15 in each group,and the balance test between groups were carried out. On the 15 th day, except for the normal group, each group was given a150 mg/kg adenine gavage maintenance model every other day, and the right group and the Guilu Erxian Cream group were intragastrically administered once a day. On the 46 th day, blood was anesthetized and sacrificed. Serum levels of creatinine, urea,glandular hormone and reproductive related hormones were detected in each group. HE staining was used to detect histopathological changes; Immunohistochemistry and immunoblotting were used to detect EPO in kidney and testis. Mediates the content of key proteins in the HIF1α-STAT5 pathway. Results: Compared with the normal group, the serum levels of creatinine and urea in the model group increased significantly, and the contents of ACTH, T3 and EPO decreased significantly(P<0.05, P<0.01). Compared with the model group, each dose group of Yougui Drink can reduce the serum creatinine and urea content in rats, increase the content of ACTH and EPO in serum(P<0.05, P<0.01), and relieve the pathological changes of rat kidney and testis. Yougui Drink can significantly regulate EPO-mediated key protein HIF1α in HIF1α-STAT5 pathway in kidney and testis expression. Conclusion: EPO is an important factor in the hypoplasia of kidney deficiency caused by adenine. The Yougui Drink has a significant reversal effect on the abnormal index of rats with hypothyroidism caused by adenine; the Yougui Drink and up-regulate the EPO content in serum, and then regulate the key protein expression of EPO-mediated HIF1α-STAT5 pathway in kidney and testis.
引文
[1]吴复苍.临床常用百方精解.天津:天津科学技术出版社,2004:215-218
[2]段富津.方剂学.上海:上海科技出版社,2010
[3]沈兴潮,宋才渊,吕帅洁,等.显微CT观察右归饮对鼠激素性股骨头坏死预防作用的实验研究.中国骨伤,2015,28(12):1106-1110
[4]范连霞,章建华,尹华,等.右归饮通过Wnt/ERK信号通路交叉串扰影响成骨细胞相关蛋白的表达.中华中医药杂志,2016,31(10):1673-1727
[5]刘逢琴,张峰,李贵海,等.右归饮对小鼠免疫功能的影响.山东中医学院学报,1996(1):43-45
[6]王杰,曾杰,许明贺,等.论脾肾两虚是男性不育症的主要病机.中华中医药杂志,2018,33(7):2866-2869
[7]Elliott S,Sinclair A,Collins H,et al.Progress in detecting cell-surface protein receptors:the erythropoietin receptor example.Ann Hematol,2014,93(2):181-192
[8]陈力,王小琴.促红细胞生成素为肾藏精理论主要物质基础之一.中华中医药学刊,2017,35(3):537-540
[9]孙广仁.中医基础理论.北京:中国中医药出版社,2002:92-94
[10]吴永生,邱山东,林求诚.中老年女性生殖激素水平与不同肾虚证型关系的研究.福建中医药,2000,31(1):3-4
[11]吴永生,叶钦勇,林求诚.中老年男性生殖激素水平与不同肾虚证型关系的研究.福建中医药,2000,31(2):3-4
[12]童骏峰,徐志伟,杨元宵,等.腺嘌呤与氢化可的松所致大鼠肾阳虚证模型比较研究.中华中医药杂志,2015,30(11):3901-3904
[13]陈俊蓉,陈利国,谢林林.关于腺嘌呤慢性肾衰实验模型的思考.实验动物科学,2013,30(2):65-67
[14]李许.EPO与“肾精”相似性的体内验证及“补肾益精”中药健脑作用机制研究.重庆:西南大学,2016
[15]李燕君.EPO与肾虚大鼠脑组织神经细胞的关系及右归饮的保护作用研究.重庆:西南大学,2015
[16]杨盛.EPO促脑神经血管单元细胞生长及补肾益精方药促EPO生成作用研究.重庆:西南大学,2016
[2]段富津.方剂学.上海:上海科技出版社,2010
[3]沈兴潮,宋才渊,吕帅洁,等.显微CT观察右归饮对鼠激素性股骨头坏死预防作用的实验研究.中国骨伤,2015,28(12):1106-1110
[4]范连霞,章建华,尹华,等.右归饮通过Wnt/ERK信号通路交叉串扰影响成骨细胞相关蛋白的表达.中华中医药杂志,2016,31(10):1673-1727
[5]刘逢琴,张峰,李贵海,等.右归饮对小鼠免疫功能的影响.山东中医学院学报,1996(1):43-45
[6]王杰,曾杰,许明贺,等.论脾肾两虚是男性不育症的主要病机.中华中医药杂志,2018,33(7):2866-2869
[7]Elliott S,Sinclair A,Collins H,et al.Progress in detecting cell-surface protein receptors:the erythropoietin receptor example.Ann Hematol,2014,93(2):181-192
[8]陈力,王小琴.促红细胞生成素为肾藏精理论主要物质基础之一.中华中医药学刊,2017,35(3):537-540
[9]孙广仁.中医基础理论.北京:中国中医药出版社,2002:92-94
[10]吴永生,邱山东,林求诚.中老年女性生殖激素水平与不同肾虚证型关系的研究.福建中医药,2000,31(1):3-4
[11]吴永生,叶钦勇,林求诚.中老年男性生殖激素水平与不同肾虚证型关系的研究.福建中医药,2000,31(2):3-4
[12]童骏峰,徐志伟,杨元宵,等.腺嘌呤与氢化可的松所致大鼠肾阳虚证模型比较研究.中华中医药杂志,2015,30(11):3901-3904
[13]陈俊蓉,陈利国,谢林林.关于腺嘌呤慢性肾衰实验模型的思考.实验动物科学,2013,30(2):65-67
[14]李许.EPO与“肾精”相似性的体内验证及“补肾益精”中药健脑作用机制研究.重庆:西南大学,2016
[15]李燕君.EPO与肾虚大鼠脑组织神经细胞的关系及右归饮的保护作用研究.重庆:西南大学,2015
[16]杨盛.EPO促脑神经血管单元细胞生长及补肾益精方药促EPO生成作用研究.重庆:西南大学,2016