真武汤及其拆方对肾小管上皮细胞氧化损伤的干预作用研究
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摘要
目的:研究真武汤及温阳组、利水组对肾小管上皮细胞氧化损伤的干预作用,为中医治则治法的科学性提供实验依据。方法:不同浓度H2O2作用于肾小管上皮细胞HK-2,诱导HK-2细胞氧化损伤模型,采用真武汤组含药血清、温阳组含药血清、以及利水组含药血清进行干预,分别检测各中药组对损伤细胞的增殖作用,对缺氧损伤相关基因HO-1、HIF-1α、生存素(s urvivin)表达的影响,以及对肾纤维化和肾小管上皮转分化相关基因转化生长因子β1(TGF-β1)、α-平滑肌肌动蛋白(SMA)、钙黏蛋白(E-c a dherin)基因表达的影响。结果:H2O2作用不同时间可造成HK-2细胞损伤明显。与模型组比较,真武汤组和利水组具有明显修复HK-2细胞生长的作用(P<0.05),温阳组作用不显著;真武汤组、温阳组及利水组均可上调HO-1、HIF-1α的表达,抑制s urvivin的表达(P<0.05),但三者之间差异不明显。真武汤组、温阳组及利水组可降低SMA及E-c a dherin的表达,温阳组可下调TGF-β1的表达,差异均有统计学意义(P<0.05)。结论:真武汤对体外HK-2细胞氧化损伤模型具有明显的保护作用,利水组对损伤模型的修复作用比较明显,温阳组抗肾纤维化及肾小管上皮转分化作用比较明显。本研究为中医治则治法的科学性提供了实验依据。
Objective: To study the intervention effects of Zhenwu tang and its decomposed recipes on the oxidative damage of renal tubular epithelial cells and to provide experimental evidence for the scientific character of principles of the Chinese medicine treatment. Methods:Used different concentrations of H2 O2 to act on renal tubular epithelial cells HK-2 to form a model of the oxidative damage in renal tubular epithelial cells. Intervened by the group of Zhenwu tang in serum containing drug, the group for warming yang in serum containing drug and the group for promoting diuresis in serum containing drug, and detected the proliferation effect in injured cells in each Chinese medicine group as well as the effects on the expressions of hypoxia-injured genes(HO-1, HIF-1α and survivin)respectively. Detected the effects on renal fibrosis and the expressions of relative genes of renal tubular epithelial transdifferentiation,such as TGF-β1,SMA and E-cadherin. Results:H2 O2 can cause significant damages to HK-2 cells at different times. Compared with the model group,the group of Zhenwu tang and the group for promoting diuresis had a significant effect in repairing HK-2 cells(P <0.05),while the group for warming yang had no obvious effect. The group of Zhenwu tang,the group for warming yang and the group for promoting diuresis can elevated the expressions of HO-1, HIF-1α genes and had a significant effect in inhibiting survivin genes.when compared with the model group(P < 0.05), but the differences among the three groups were not evident. The group of Zhenwu tang,the group of warming yang and the group for promoting diuresis can reduce the expressions of SMA and E-cadherin genes;and the group of warming yang can reduce the expressions of TGF-β1,differences being significant(P < 0.05). Conclusion:Zhenwu tang has an obvious effect in protecting the model of oxidative damage renal tubular epithelial cells in vitro. the group for promoting diuresishas a more significant effect in repairing the injured model and the group for warming yang has an evident effect in ameliorating renal fibrosis and renal tubular epithelial transdifferentiation. This study provides an experimental basis for the scientific character of principles of Chinese medicine treatment.
Objective: To study the intervention effects of Zhenwu tang and its decomposed recipes on the oxidative damage of renal tubular epithelial cells and to provide experimental evidence for the scientific character of principles of the Chinese medicine treatment. Methods:Used different concentrations of H2 O2 to act on renal tubular epithelial cells HK-2 to form a model of the oxidative damage in renal tubular epithelial cells. Intervened by the group of Zhenwu tang in serum containing drug, the group for warming yang in serum containing drug and the group for promoting diuresis in serum containing drug, and detected the proliferation effect in injured cells in each Chinese medicine group as well as the effects on the expressions of hypoxia-injured genes(HO-1, HIF-1α and survivin)respectively. Detected the effects on renal fibrosis and the expressions of relative genes of renal tubular epithelial transdifferentiation,such as TGF-β1,SMA and E-cadherin. Results:H2 O2 can cause significant damages to HK-2 cells at different times. Compared with the model group,the group of Zhenwu tang and the group for promoting diuresis had a significant effect in repairing HK-2 cells(P <0.05),while the group for warming yang had no obvious effect. The group of Zhenwu tang,the group for warming yang and the group for promoting diuresis can elevated the expressions of HO-1, HIF-1α genes and had a significant effect in inhibiting survivin genes.when compared with the model group(P < 0.05), but the differences among the three groups were not evident. The group of Zhenwu tang,the group of warming yang and the group for promoting diuresis can reduce the expressions of SMA and E-cadherin genes;and the group of warming yang can reduce the expressions of TGF-β1,differences being significant(P < 0.05). Conclusion:Zhenwu tang has an obvious effect in protecting the model of oxidative damage renal tubular epithelial cells in vitro. the group for promoting diuresishas a more significant effect in repairing the injured model and the group for warming yang has an evident effect in ameliorating renal fibrosis and renal tubular epithelial transdifferentiation. This study provides an experimental basis for the scientific character of principles of Chinese medicine treatment.
引文
[1]李莎莎,韩凌,黎莉,等.真武汤对肾纤维化大鼠尿蛋白含量及血生化指标的影响[J].江西中医药大学学报,2011,23(2):64-67.
[2]李莎莎,肖雪,韩凌,等.真武汤对肾纤维化大鼠血清和肾脏组织中SOD活力、MDA含量的影响[J].中药药理与临床,2012,28(2):18-20.
[3]韩凌,李莎莎,冯森玲,等.真武汤对大鼠单侧输尿管梗阻模型细胞因子的影响[J].中国实验方剂学杂志,2012,18(3):164-168.
[4]侯兆辉,侯志敏.《伤寒论》真武汤方证浅析[J].中国中医急症,2010,19(3):487.
[5]韩凌,冯森玲,李莎莎,等.真武汤对单侧输尿管梗阻模型大鼠尿KIM-1、clusterin、OPN含量的影响[J].中药药理与临床,2012,28(1):13-17.
[6]毕礼明,陈英兰,马济佩.中西医结合治疗慢性肾脏病的切入点探讨[J].中华中医药学刊,2009,28(8):1759-1760.
[7]鞠静,杜武勋.真武汤药效物质基础及温阳利水机制研究[J].吉林中医药,2016,36(7):719-723.
[8]邓晓红,黄建华,董竞成.附子药理作用的分子机制研究进展[J].江西中医药大学学报,2018,30(1):121-124.
[9]游昕,熊大国,郭志斌,等.茯苓多种化学成分及药理作用的研究进展[J].安徽农业科学,2015,43(2):106-109.
[10]王小飞,吴国泰,牛亭惠,等.生姜的化学、药理及应用[J].中国果菜,2016,36(6):23-26.
[11]王涵,杨娜,谭静,等.白术化学成分、药理作用及临床应用的研究进展[J].甘肃医药,2018,37(1):23-26.
[12]金英善,陈曼丽,陶俊.芍药化学成分和药理作用研究进展[J].中国药理学与毒理学杂志,2013,27(4):745-750.
[13]陈海燕.急性肾损伤的生物标志物—血红素加氧酶[J].天津医科大学学报,2017,23(5):483-484.
[14]Zager RA,Johnson AC,Becker K.Plasma and urinary heme oxygenase-1 in AKI[J].Journal of the American Society of Nephrology Jasn,2012,23(6):1048-1057.
[15]唐群,吴华,张熙,等.六味地黄汤对慢性肾衰模型大鼠PHD2和HIF-1α表达的影响[J].北京中医药大学学报,2017,40(4):290-296.
[16]苗龙,刘洋,刘林,等.肾细胞癌组织中PTEN和HIF-1α的表达及临床意义[J].现代生物医学进展,2017,17(6):1127-1130.
[17]汪峰,张斌斌,郭巍,等.舒尼替尼对转移性肾细胞癌患者临床疗效及生存素、端粒酶表达的影响[J].现代医学,2017,45(4):552-558.
[18]周颖,赵敏,张辉,等.缺氧诱导因子-1α和Survivin蛋白在非小细胞肺癌中的表达及其与人乳头状瘤病毒感染的相关性[J].中国老年学,2017,37(6):1406-1408.
[19]孙伟,尹雪皎,涂玥,等.芪附汤调节ERK1/2信号通路改善UUO阳虚证模型肾间质纤维化的作用和机制[J].中国中药杂志,2014,39(21):4082-4089.
[20]汪洋,牟建军,刘富强,等.高盐喂食对Dahl盐敏感大鼠肾小管上皮向间质转化和肾脏纤维化的影响[J].中华高血压杂志,2015,32(3):1524-1525.
[21]潘祉谕,达静静,董蓉,等.Snail1/IGF-1信号通路介导高糖诱导的肾小管上皮细胞EMT[J].中国病理生理杂志,2017,33(9):1662-1668.
[22]Yang J,Liu Y.Dissection of key events in tubular epithelial to myofibroblast transition and its implications in renal interstitial fibrosis[J].American Journal of Pathology,2001,159(4):1465-1475.
[23]蔡月琴,褚燕青,朱科燕,等.mi R-21对TGF-β1诱导的肾小管上皮细胞间质转分化的影响[J].中国应用生理学杂志,2017,33(4):346-350.
[2]李莎莎,肖雪,韩凌,等.真武汤对肾纤维化大鼠血清和肾脏组织中SOD活力、MDA含量的影响[J].中药药理与临床,2012,28(2):18-20.
[3]韩凌,李莎莎,冯森玲,等.真武汤对大鼠单侧输尿管梗阻模型细胞因子的影响[J].中国实验方剂学杂志,2012,18(3):164-168.
[4]侯兆辉,侯志敏.《伤寒论》真武汤方证浅析[J].中国中医急症,2010,19(3):487.
[5]韩凌,冯森玲,李莎莎,等.真武汤对单侧输尿管梗阻模型大鼠尿KIM-1、clusterin、OPN含量的影响[J].中药药理与临床,2012,28(1):13-17.
[6]毕礼明,陈英兰,马济佩.中西医结合治疗慢性肾脏病的切入点探讨[J].中华中医药学刊,2009,28(8):1759-1760.
[7]鞠静,杜武勋.真武汤药效物质基础及温阳利水机制研究[J].吉林中医药,2016,36(7):719-723.
[8]邓晓红,黄建华,董竞成.附子药理作用的分子机制研究进展[J].江西中医药大学学报,2018,30(1):121-124.
[9]游昕,熊大国,郭志斌,等.茯苓多种化学成分及药理作用的研究进展[J].安徽农业科学,2015,43(2):106-109.
[10]王小飞,吴国泰,牛亭惠,等.生姜的化学、药理及应用[J].中国果菜,2016,36(6):23-26.
[11]王涵,杨娜,谭静,等.白术化学成分、药理作用及临床应用的研究进展[J].甘肃医药,2018,37(1):23-26.
[12]金英善,陈曼丽,陶俊.芍药化学成分和药理作用研究进展[J].中国药理学与毒理学杂志,2013,27(4):745-750.
[13]陈海燕.急性肾损伤的生物标志物—血红素加氧酶[J].天津医科大学学报,2017,23(5):483-484.
[14]Zager RA,Johnson AC,Becker K.Plasma and urinary heme oxygenase-1 in AKI[J].Journal of the American Society of Nephrology Jasn,2012,23(6):1048-1057.
[15]唐群,吴华,张熙,等.六味地黄汤对慢性肾衰模型大鼠PHD2和HIF-1α表达的影响[J].北京中医药大学学报,2017,40(4):290-296.
[16]苗龙,刘洋,刘林,等.肾细胞癌组织中PTEN和HIF-1α的表达及临床意义[J].现代生物医学进展,2017,17(6):1127-1130.
[17]汪峰,张斌斌,郭巍,等.舒尼替尼对转移性肾细胞癌患者临床疗效及生存素、端粒酶表达的影响[J].现代医学,2017,45(4):552-558.
[18]周颖,赵敏,张辉,等.缺氧诱导因子-1α和Survivin蛋白在非小细胞肺癌中的表达及其与人乳头状瘤病毒感染的相关性[J].中国老年学,2017,37(6):1406-1408.
[19]孙伟,尹雪皎,涂玥,等.芪附汤调节ERK1/2信号通路改善UUO阳虚证模型肾间质纤维化的作用和机制[J].中国中药杂志,2014,39(21):4082-4089.
[20]汪洋,牟建军,刘富强,等.高盐喂食对Dahl盐敏感大鼠肾小管上皮向间质转化和肾脏纤维化的影响[J].中华高血压杂志,2015,32(3):1524-1525.
[21]潘祉谕,达静静,董蓉,等.Snail1/IGF-1信号通路介导高糖诱导的肾小管上皮细胞EMT[J].中国病理生理杂志,2017,33(9):1662-1668.
[22]Yang J,Liu Y.Dissection of key events in tubular epithelial to myofibroblast transition and its implications in renal interstitial fibrosis[J].American Journal of Pathology,2001,159(4):1465-1475.
[23]蔡月琴,褚燕青,朱科燕,等.mi R-21对TGF-β1诱导的肾小管上皮细胞间质转分化的影响[J].中国应用生理学杂志,2017,33(4):346-350.