复方鳖甲软肝片对小鼠肾纤维化的防治作用
|
摘要
目的:观察复方鳖甲软肝片对小鼠肾纤维化的防治作用。方法:40只昆明种小鼠随机分为对照组、模型组、治疗组和干预组,模型组、治疗组和干预组皮下注射CCl4与橄榄油混合液持续6周建立肾纤维化动物模型,干预组于造模同时开始给予复方鳖甲软肝片干预,共12周,对照组代以等体积橄榄油皮下注射;造模后治疗组复方鳖甲软肝片连续灌胃给药6周,模型组和对照组则予等体积生理盐水灌胃;实验第12周时检测血清尿素氮(BUN)、肌酐(Scr)水平,ELISA法检测血清透明质酸(HA)、层黏连蛋白(LN)、Ⅲ型前胶原(PC III)、Ⅳ型胶原(Ⅳ-Col),采用HE染色和Masson染色评价肾间质纤维化损伤程度,免疫组织化学染色检测肾脏组织α-平滑肌动蛋白(α-SMA)和肿瘤坏死因子(TNF-α)表达。结果:与对照组比较,模型组血清BUN、Scr、HA、LN、PC III、Ⅳ-Col均明显增高(P <0. 05),肾间质纤维化明显,肾组织α-SMA和TNF-α高表达(P <0. 05);与模型组比较,治疗组和干预组上述肾功能和肾纤维化指标均显著降低(P <0. 05),肾间质纤维化改善,肾组织中α-SMA和TNF-α表达降低(P <0. 05)。结论:复方鳖甲软肝片能有效减轻并预防CCl4诱导的小鼠肾纤维化及肾脏损害。
Objective: To observe the preventive effect and treatment of compound Biejiaruangan troche on renal fibrosis in mice. Methods: 40 Kunming category rats were divided randomly into four groups: control group,model group,treatment group and intervention group. Model group,treatment group and intervention group were given subcutaneous injection of equal volume mixture of CCl4 and olive oil for 6 weeks to establish renal fibrosis animal model,intervention group was given compound Biejiaruangan troche for 12 weeks; at the same time,the control group was injected subcutaneously with equal volume of olive oil. The treatment group was given intragastric compound Biejiaruangan troche for 6 weeks after model establishment. Model and control groups were given equal volume of physiological saline by gavage. Serum blood urea nitrogen( BUN) and serum creatinine( Scr) were detected; hyaluronic acid( HA),laminin( LN),procollagen III( PC III) and type IV collagen( Ⅳ-Col) were detected by Enzyme-linked immunosorbent assay( ELISA). Renal tubular interstitial fibrosis was evaluated by HE and Masson staining. The protein expression levels of α-SMA and TNF-α were detected by immunohistochemical method. Results: Compared with the control group,the levels of BUN,Scr,HA,LN,PC III,type IV collagen obviously improved( P < 0. 05),renal tubular interstitial fibrosis was obvious,expression levels of α-SMA and TNF-α increased in the renal tissue( P <0. 05). Compared with the model group,the treatment group and intervention group showed obvious decreased levels of renal function and renal fibrosis index( P < 0. 05),the renal interstitial fibrosis was improved,and protein expression levels of α-SMA and TNF-α decreased( P < 0. 05). Conclusion: The compound Biejiaruangan troche could effectively reduce and prevent CCl4 induced renal fibrosis and renal damage.
Objective: To observe the preventive effect and treatment of compound Biejiaruangan troche on renal fibrosis in mice. Methods: 40 Kunming category rats were divided randomly into four groups: control group,model group,treatment group and intervention group. Model group,treatment group and intervention group were given subcutaneous injection of equal volume mixture of CCl4 and olive oil for 6 weeks to establish renal fibrosis animal model,intervention group was given compound Biejiaruangan troche for 12 weeks; at the same time,the control group was injected subcutaneously with equal volume of olive oil. The treatment group was given intragastric compound Biejiaruangan troche for 6 weeks after model establishment. Model and control groups were given equal volume of physiological saline by gavage. Serum blood urea nitrogen( BUN) and serum creatinine( Scr) were detected; hyaluronic acid( HA),laminin( LN),procollagen III( PC III) and type IV collagen( Ⅳ-Col) were detected by Enzyme-linked immunosorbent assay( ELISA). Renal tubular interstitial fibrosis was evaluated by HE and Masson staining. The protein expression levels of α-SMA and TNF-α were detected by immunohistochemical method. Results: Compared with the control group,the levels of BUN,Scr,HA,LN,PC III,type IV collagen obviously improved( P < 0. 05),renal tubular interstitial fibrosis was obvious,expression levels of α-SMA and TNF-α increased in the renal tissue( P <0. 05). Compared with the model group,the treatment group and intervention group showed obvious decreased levels of renal function and renal fibrosis index( P < 0. 05),the renal interstitial fibrosis was improved,and protein expression levels of α-SMA and TNF-α decreased( P < 0. 05). Conclusion: The compound Biejiaruangan troche could effectively reduce and prevent CCl4 induced renal fibrosis and renal damage.
引文
[1]PEI G,ZENG R,HAN M R et al. Renal interstitial infil-tration and tertiary lymphoid organ neogenesis in Ig A ne-phropathy[J]. Clin J Am Soc Nephrol,2014,9(2):255-264.
[2]SNYDER J J,FOLEY R N,COLLINS A J. Prevalence ofCKD in the United States:a sensitivity analysis using theNational Health and Nutrition Examination Survey(NHANES)1999-2004[J]. Am J Kidney Dis,2009,53(2):218-228.
[3]余红霞.鳖甲软肝片在血吸虫病肝纤维化患者中的应用价值研究[J].世界中西医结合杂志,2015,10(9):1274-1275.
[4]申定珠,陶庆,胡义扬,等.四氯化碳肝纤维化大鼠肝组织差异蛋白质组的动态变化[J].中华肝脏病杂志,2012,20(9):664-670.
[5]TRUONG N H,NYUYEN N H,NGUYEN N K T,et al.Establishment of a standardized mouse model of hepatic fi-brosis for biomedical research[J]. Biomed Res Ther,2014,1(2):43-49.
[6]陈凤,戴恩来.中西医对肾纤维化发病机制的研究进展[J].辽宁中医杂志,2016,43(6):1243-1246.
[7]NOGUEIRA A,PIRES M J,OLIVEIRA P A. Pathophys-iological mechanisms of renal fibrosis:a review of animalmodels and therapeutic strategies[J]. In Vivo,2017,31(1):1-22.
[8]贺峥,张磊,金晓明.肾纤维化发生的细胞及分子生物学机制研究进展[J].国际免疫学杂志,2016,39(5):488-492.
[9]陈建,曾莉,陈刚,等.抗纤灵方对AngⅡ诱导肾纤维化小鼠肾组织α-SMA和Ⅰ型胶原的影响[J].中华中医药杂志,2017,32(2):739-742.
[10]王天生,王东红,师军华.慢性肾脏病患者血液透析前后肾纤维化指标及肾功能的变化研究[J].中国实验诊断学,2016,20(7):1090-1093.
[11]徐敏,项荣.哌唑嗪对糖尿病肾病大鼠肾纤维化与基质金属蛋白酶表达水平的影响[J].中国临床药理学杂志,2018,34(21):2532-2535.
[12]余彦霖,肖堂利,刘亮,等.硫酸吲哚酚促进肾纤维化的机制研究[J].第三军医大学学报,2016,(2):119-123.
[13]李林蔚,周景华,刘华生.柴胡桂枝汤对肝纤维化大鼠FN、a-SMA表达的影响[J].黑龙江医药,2014,27(2):265-267.
[14]陈晛,何立群.健脾清化方在肾小球硬化大鼠中抗肾纤维化的作用及其机制[J].中国医学科学院学报,2014,36(5):461-465.
[15]李均,陈雨思,邹朝霞.丹酚酸A、B组分配伍对HK-2细胞CCL2、CXCL10及氧化应激反应的影响[J].山东医药,2016,56(46):7-10.
[16]SHIMA H,SASAKI K,SUZUKI T,et al. A novel indolecompound MA-35 attenuates renal fibrosis by inhibitingboth TNF-αand TGF-β1pathways.[J]Sci Rep,2017,7(1):1884-1889.
[17]周瑾,陈香美,魏日胞,等.复方鳖甲软肝片方对TGF-β1诱导的肾间质成纤维细胞增殖及分泌细胞外基质的影响[J].中国中西医结合肾病杂志,2014,15(5):381-384.
[18]钟建,赵宁博.肾纤维化络病实质及辨治探讨[J].南京中医药大学学报,2014,30(6):510-512.
[2]SNYDER J J,FOLEY R N,COLLINS A J. Prevalence ofCKD in the United States:a sensitivity analysis using theNational Health and Nutrition Examination Survey(NHANES)1999-2004[J]. Am J Kidney Dis,2009,53(2):218-228.
[3]余红霞.鳖甲软肝片在血吸虫病肝纤维化患者中的应用价值研究[J].世界中西医结合杂志,2015,10(9):1274-1275.
[4]申定珠,陶庆,胡义扬,等.四氯化碳肝纤维化大鼠肝组织差异蛋白质组的动态变化[J].中华肝脏病杂志,2012,20(9):664-670.
[5]TRUONG N H,NYUYEN N H,NGUYEN N K T,et al.Establishment of a standardized mouse model of hepatic fi-brosis for biomedical research[J]. Biomed Res Ther,2014,1(2):43-49.
[6]陈凤,戴恩来.中西医对肾纤维化发病机制的研究进展[J].辽宁中医杂志,2016,43(6):1243-1246.
[7]NOGUEIRA A,PIRES M J,OLIVEIRA P A. Pathophys-iological mechanisms of renal fibrosis:a review of animalmodels and therapeutic strategies[J]. In Vivo,2017,31(1):1-22.
[8]贺峥,张磊,金晓明.肾纤维化发生的细胞及分子生物学机制研究进展[J].国际免疫学杂志,2016,39(5):488-492.
[9]陈建,曾莉,陈刚,等.抗纤灵方对AngⅡ诱导肾纤维化小鼠肾组织α-SMA和Ⅰ型胶原的影响[J].中华中医药杂志,2017,32(2):739-742.
[10]王天生,王东红,师军华.慢性肾脏病患者血液透析前后肾纤维化指标及肾功能的变化研究[J].中国实验诊断学,2016,20(7):1090-1093.
[11]徐敏,项荣.哌唑嗪对糖尿病肾病大鼠肾纤维化与基质金属蛋白酶表达水平的影响[J].中国临床药理学杂志,2018,34(21):2532-2535.
[12]余彦霖,肖堂利,刘亮,等.硫酸吲哚酚促进肾纤维化的机制研究[J].第三军医大学学报,2016,(2):119-123.
[13]李林蔚,周景华,刘华生.柴胡桂枝汤对肝纤维化大鼠FN、a-SMA表达的影响[J].黑龙江医药,2014,27(2):265-267.
[14]陈晛,何立群.健脾清化方在肾小球硬化大鼠中抗肾纤维化的作用及其机制[J].中国医学科学院学报,2014,36(5):461-465.
[15]李均,陈雨思,邹朝霞.丹酚酸A、B组分配伍对HK-2细胞CCL2、CXCL10及氧化应激反应的影响[J].山东医药,2016,56(46):7-10.
[16]SHIMA H,SASAKI K,SUZUKI T,et al. A novel indolecompound MA-35 attenuates renal fibrosis by inhibitingboth TNF-αand TGF-β1pathways.[J]Sci Rep,2017,7(1):1884-1889.
[17]周瑾,陈香美,魏日胞,等.复方鳖甲软肝片方对TGF-β1诱导的肾间质成纤维细胞增殖及分泌细胞外基质的影响[J].中国中西医结合肾病杂志,2014,15(5):381-384.
[18]钟建,赵宁博.肾纤维化络病实质及辨治探讨[J].南京中医药大学学报,2014,30(6):510-512.