基于玄府理论的固本通络方对IgA肾病大鼠Podocin mRNA和α-actinin-4 mRNA表达的影响
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摘要
目的:探析基于玄府理论的固本通络方对IgA肾病大鼠Podocin mRNA和α-actinin-4 mRNA表达的调节及对IgA肾病蛋白尿的治疗作用。方法:将80只SD大鼠随机分为正常组、模型组、中药组和科素亚组,采用牛血清清蛋白+四氯化碳+蓖麻油+脂多糖的方法建立IgA肾病大鼠模型。分别于造模结束后、治疗2周、治疗4周后检测各组大鼠24 h尿蛋白定量、血肌酐、尿素氮水平。治疗2周和4周时,采用光镜、免疫荧光法观察肾小球形态学变化;PCR法检测Prodocin、α-actinin-4 mRNA表达量。结果:治疗前,正常组24 h尿蛋白定量明显少于模型组,差异具有统计学意义(P<0.05)。模型组、中药组及科素亚组肾脏病理示肾小球系膜增生,基底膜增厚,系膜区IgA荧光强度极高。经治疗,与模型组比较,中药组及科素亚组在治疗2周、4周后24 h尿蛋白定量明显下降,差异具有统计学意义(P<0.05);中药组在治疗2周、治疗4周后与科素亚组比较24 h尿蛋白定量均减少,差异具有统计学意义(P<0.05)。与模型组比较,治疗后中药组和科素亚组Podocin mRNA含量明显升高,α-actinin-4 mRNA含量均明显降低,差异具有统计学意义(P<0.05);治疗4周与治疗2周比较,科素亚组Podocin含量明显升高,中药组α-actinin-4 mRNA含量均明显降低,差异具有统计学意义(P<0.01);治疗4周后中药组α-actinin-4 mRNA含量均明显低于科素亚组,差异具有统计学意义(P<0.05)。中药组及科素亚组肾小球病变较治疗前损伤程度轻,荧光强度明显减低。结论:固本通络方可以减轻IgA肾病大鼠蛋白尿与调节足细胞Podocin和α-actinin-4表达有关。
Objective:To explore the effects of the Guben Tongluo Formula on the expression of Podocin mRNA and alpha-actinin-4 mRNA in IgA nephropat hy rats and the treatment of urinary protein in IgA nephropathy. Methods:Eighty SD rats were randomly divided into four goups:control group,model group,herb group and Cozaar group,with the method of Bovine serum albumin + carbon tetrachloride + castor oil + lipopolysaccharide establishing rat model of IgA nephropathy. The 24-hour urine protein quantitation,serum creatinine and urea nitrogen levels of each group were measured after the end of modeling,2 weeks and 4 weeks. At the end of 2 nd and 4 th week,the morphological changes of glomeruli were observed by light microscopy and immunofluorescence and PCR was uesd respectively to examine the mRNA expression of Podocin and α-actinin-4. Results:Before treatment,the 24-hour urine protein in the normal group was significantly less than that in the model group,which was statistically significant(P< 0.05). Renal pathology of the model group,Chinese medicine group and Kosuya group showed mesangial hyperplasia,thickening of the basement membrane,and high fluorescence intensity of IgA in the mesangial area. After treatment,at the end of 2 nd and 4 th week,the urine protein quantitation of the Chinese medicine group and Kosuya group was significantly lower than that in the model group,with statistical significance(P< 0.05). At the end of 2 nd and 4 th week,compared with the Cozaar group,the urinary protein quantitation of the Chinese medicine group was reduced and there was a statisti cal significance(P< 0.05). Compared with the model group,the content of Podocin mRNA of the Chinese medicine group and Kosuya group was significantly increased,and the content of α-actinin-4 mRNA was significantly decreased after the treatment(P< 0.05). At the end of 4 th week,compared with the results of treatment for 2 nd weeks,the content of Podocin in the Kossuya group was significantly increased,and the content of α-actinin-4 mRNA in the Chinese medicine group was significantly decreased(P< 0.01). The content of α-actinin-4 mRNA in the Chinese medicine group was significantly lower than that in the Kossuya group at the end of 4 th week(P< 0.05). The glomerular lesions in the Chinese medicine group and Kosuya group were lighter than before treatment,and the fluorescence intensity was significantly reduced. Conclusion:Guben TongLuo Formula can alleviate proteinuria in rats with IgA nephropathy,it is associated with the regulation of Podocin and α-actinin-4 expression in podocytes.
Objective:To explore the effects of the Guben Tongluo Formula on the expression of Podocin mRNA and alpha-actinin-4 mRNA in IgA nephropat hy rats and the treatment of urinary protein in IgA nephropathy. Methods:Eighty SD rats were randomly divided into four goups:control group,model group,herb group and Cozaar group,with the method of Bovine serum albumin + carbon tetrachloride + castor oil + lipopolysaccharide establishing rat model of IgA nephropathy. The 24-hour urine protein quantitation,serum creatinine and urea nitrogen levels of each group were measured after the end of modeling,2 weeks and 4 weeks. At the end of 2 nd and 4 th week,the morphological changes of glomeruli were observed by light microscopy and immunofluorescence and PCR was uesd respectively to examine the mRNA expression of Podocin and α-actinin-4. Results:Before treatment,the 24-hour urine protein in the normal group was significantly less than that in the model group,which was statistically significant(P< 0.05). Renal pathology of the model group,Chinese medicine group and Kosuya group showed mesangial hyperplasia,thickening of the basement membrane,and high fluorescence intensity of IgA in the mesangial area. After treatment,at the end of 2 nd and 4 th week,the urine protein quantitation of the Chinese medicine group and Kosuya group was significantly lower than that in the model group,with statistical significance(P< 0.05). At the end of 2 nd and 4 th week,compared with the Cozaar group,the urinary protein quantitation of the Chinese medicine group was reduced and there was a statisti cal significance(P< 0.05). Compared with the model group,the content of Podocin mRNA of the Chinese medicine group and Kosuya group was significantly increased,and the content of α-actinin-4 mRNA was significantly decreased after the treatment(P< 0.05). At the end of 4 th week,compared with the results of treatment for 2 nd weeks,the content of Podocin in the Kossuya group was significantly increased,and the content of α-actinin-4 mRNA in the Chinese medicine group was significantly decreased(P< 0.01). The content of α-actinin-4 mRNA in the Chinese medicine group was significantly lower than that in the Kossuya group at the end of 4 th week(P< 0.05). The glomerular lesions in the Chinese medicine group and Kosuya group were lighter than before treatment,and the fluorescence intensity was significantly reduced. Conclusion:Guben TongLuo Formula can alleviate proteinuria in rats with IgA nephropathy,it is associated with the regulation of Podocin and α-actinin-4 expression in podocytes.
引文
[1]王悦,赵明辉,章友康. IgA肾病分子病因学的研究[J].肾脏病与透析肾移植杂志,2002,11(3):273-276.
[2] IAMES V,JOSEPH P. IgA nephropathy[J]. N Engl J Med,2002,347(10):738-748.
[3]齐尔,蒋更如. IgA肾病遗传学的研究进展[J].医学综述,2012,18(20):3437-3440.
[4] GOTO M,WAKAI K,KAWAMURA T,et al. A scorlng system to predict renaloutcome in IgA nephropathy:a nationwide 10-year prospective cohortstudy[J]. Nephrol Dial Transplant,2009,24(10):3068-3074.
[5] REICH H N,TROYANOV S,SCHOLEY J W,et al. Predicting the risk for dialysis or death in IgA nephropathy[J]. J Am Soc Nephrol,2011,22(4):752-761.
[6] HARA M,YANAGIHARA T,KIHARA I. Cumulative excretion of urinary podocytes reflects disease progression in IgA nephropathy and Schênlein-Henoch purpuranephritis[J]. Clin J Am Soc Nephrol,2007,2(2):231-238.
[7]瞿中洁,朱彩凤,朱斌. IgA肾病中足细胞损伤机制的进展研究[J].中国中西医结合肾病杂志,2011,12(6):563-564
[8] ZHANG C,JING H J,CHANG Y,et al. Downregulating of CD2 associated protein imparied the physiological functions of podocytes[J]. Cell Biology International,2009,33(6):623-639.
[9]汤颖,娄探奇,成彩联,等.实验性IgA肾病模型的改进[J].中山大学学报,2006,29(12):184-187.
[10] COPPO R,D AMICO G. Factors predicting progression of IgA nephropathies[J]. Nephrol,2005,18(5):503-512.
[11] EIRO M,KATOH T,KURIKI M,et al. The product of druation and proteinuria(proteinuria index)is a possible marker for glomerlar and tubulointerstitial damage in IgA nephropathy[J]. Nephron,2002,90(4):432-441
[12]李晗笑,王艳秋,王博文,等.尿液足细胞排泄与IgA肾病的相关关系[J].中国医科大学学报,2010,39(4):293-295.
[13] LIU H F,GUO L Q,ZHUANG Y Y,et al. Thiazolidinedione attenuate proteinuria and glomerulosclerosis in Adriamycin-induced nephropathy rats via slit diaphragm protection[J]. Nephrology,2010,15(1):75-83.
[14] HUBER T B,KOTTGEN M,SCHILLING B,et al. Interaction with podocin facil-itatesnephrin signaling[J]. J Biol Chem,2001,276(45):41543-41546.
[15]郑平东,周家俊,高建东,等.固本通络冲剂治疗IgA肾病的临床疗效观察[J].中国中西医结合肾病杂志,2003,4(3):150-152.
[16]黄文强,彭宁静,何利黎,等.肝玄府学说理论初探[J].中医杂志,2012,53(11):901-902.
[17]吕仕超.经皮冠状动脉介入术后无复流现象与玄府开闭的研究[J].中医药现代化,2012,14(5):1981-1984.
[18]李雯雯,黄迪,沈沛成,等.中药固本通络方对IgA肾病小鼠氧化应激作用机制的实验研究[J].四川大学学报(医学版),2017,48(2):210-215.
[2] IAMES V,JOSEPH P. IgA nephropathy[J]. N Engl J Med,2002,347(10):738-748.
[3]齐尔,蒋更如. IgA肾病遗传学的研究进展[J].医学综述,2012,18(20):3437-3440.
[4] GOTO M,WAKAI K,KAWAMURA T,et al. A scorlng system to predict renaloutcome in IgA nephropathy:a nationwide 10-year prospective cohortstudy[J]. Nephrol Dial Transplant,2009,24(10):3068-3074.
[5] REICH H N,TROYANOV S,SCHOLEY J W,et al. Predicting the risk for dialysis or death in IgA nephropathy[J]. J Am Soc Nephrol,2011,22(4):752-761.
[6] HARA M,YANAGIHARA T,KIHARA I. Cumulative excretion of urinary podocytes reflects disease progression in IgA nephropathy and Schênlein-Henoch purpuranephritis[J]. Clin J Am Soc Nephrol,2007,2(2):231-238.
[7]瞿中洁,朱彩凤,朱斌. IgA肾病中足细胞损伤机制的进展研究[J].中国中西医结合肾病杂志,2011,12(6):563-564
[8] ZHANG C,JING H J,CHANG Y,et al. Downregulating of CD2 associated protein imparied the physiological functions of podocytes[J]. Cell Biology International,2009,33(6):623-639.
[9]汤颖,娄探奇,成彩联,等.实验性IgA肾病模型的改进[J].中山大学学报,2006,29(12):184-187.
[10] COPPO R,D AMICO G. Factors predicting progression of IgA nephropathies[J]. Nephrol,2005,18(5):503-512.
[11] EIRO M,KATOH T,KURIKI M,et al. The product of druation and proteinuria(proteinuria index)is a possible marker for glomerlar and tubulointerstitial damage in IgA nephropathy[J]. Nephron,2002,90(4):432-441
[12]李晗笑,王艳秋,王博文,等.尿液足细胞排泄与IgA肾病的相关关系[J].中国医科大学学报,2010,39(4):293-295.
[13] LIU H F,GUO L Q,ZHUANG Y Y,et al. Thiazolidinedione attenuate proteinuria and glomerulosclerosis in Adriamycin-induced nephropathy rats via slit diaphragm protection[J]. Nephrology,2010,15(1):75-83.
[14] HUBER T B,KOTTGEN M,SCHILLING B,et al. Interaction with podocin facil-itatesnephrin signaling[J]. J Biol Chem,2001,276(45):41543-41546.
[15]郑平东,周家俊,高建东,等.固本通络冲剂治疗IgA肾病的临床疗效观察[J].中国中西医结合肾病杂志,2003,4(3):150-152.
[16]黄文强,彭宁静,何利黎,等.肝玄府学说理论初探[J].中医杂志,2012,53(11):901-902.
[17]吕仕超.经皮冠状动脉介入术后无复流现象与玄府开闭的研究[J].中医药现代化,2012,14(5):1981-1984.
[18]李雯雯,黄迪,沈沛成,等.中药固本通络方对IgA肾病小鼠氧化应激作用机制的实验研究[J].四川大学学报(医学版),2017,48(2):210-215.