不同药性祛风通络方干预下UUO大鼠肾小管上皮细胞TRPV4变化规律研究
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槲皮素抗肾小管上皮细胞转分化的机制研究
程弓;陈泽君;王平
【期刊名称】《西部医学》
【年(卷),期】2010(0)2
【摘要】目的探讨槲皮素对转化生长因子β(TGF-β)诱导大鼠肾小管上皮细胞-肌成纤维细胞转分化(TEMT)的影响.方法 TGF-β刺激体外培养的正常大鼠肾小管上皮细胞株(NRK52E),同时以不同浓度槲皮素进行干预,细胞免疫化学染色法和PCR 方法检测α-平滑肌肌动蛋白(α-SMA)及E-钙黏蛋白(E-cadherin)的表达.结果TGF-β刺激导致α-SMA表达增加,不同浓度槲皮素作用后,可减轻TGF-β诱导的α-SMA表达,上调E-cadherin的表达(P<0.05).结论槲皮素对TGF-β刺激的NRK52E细胞转分化有明显抑制作用.
【总页数】3页(P214-216)
【作者】程弓;陈泽君;王平
【作者单位】成都市第三人民医院肾内科,四川,成都,610031;成都市第三人民医院肾内科,四川,成都,610031;成都市第三人民医院肾内科,四川,成都,610031
【正文语种】中文
【中图分类】R285.6
【相关文献】
1.肾小管上皮细胞转分化在肾间质纤维化过程中的作用及其调控机制研究进展 [J], 罗健华
2.miR-155对肾小管上皮细胞转分化的影响及其机制研究 [J], 宁雅娴;王俭勤;王晓元
3.内皮素1在肾小管上皮细胞转分化中的作用机制研究进展 [J], 刘红;胡洪贞;李伟
4.中药抗肾小管上皮细胞间充质转分化的临床研究进展 [J], 金丽霞; 金丽军; 张佩青
5.吡格列酮对高糖诱导的大鼠肾小管上皮细胞-间充质细胞转分化的影响及机制研究 [J], 孙兰;张帆;石明隽;田平平;郭兵
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血管紧张素Ⅱ诱导肾小管上皮细胞Toll样受体4和炎症因子表达汤天凤;唐荣;蒋春明;夏阳阳;敖翔;周巧玲【期刊名称】《中国组织化学与细胞化学杂志》【年(卷),期】2016(025)004【摘要】目的探讨Toll样受体4(Toll-like receptor4,TLR4)在血管紧张素Ⅱ (AngⅡ)诱导的大鼠肾小管上皮细胞(NRK-52E)所模拟的高血压肾损害微环境中的作用机制.方法 NRK-52E细胞按AngⅡ浓度梯度(10-6~10-10mol/L)培养24h以确定最佳干预浓度,按时间梯度(6 ~48h)分组培养选择最佳干预时间;同时建立稳定转染TLR4特异RNAi质粒的NRK-52E细胞株.免疫细胞化学及RT-PCR检测TLR4表达水平,ELISA检测上清IL-6及TNF-α水平.结果 10-6 ~ 10-9mol/L的AngⅡ均可诱导NRK-52E细胞中TLR4 mRNA明显上调,其中10-7 mol/L组作用更为显著;6h即可见TLR4 mRNA水平显著增高,高峰维持12 ~24h;同时IL-6、TNF-α表达亦上调.TLR4特异性RNA干扰可显著抑制NRK-52E中TLR4的mRNA表达,逆转AngⅡ对IL-6、TNF-α表达的上调作用.结论AngⅡ可诱导NRK-52E细胞中TLR4及IL-6、TNF-α的表达,阻断TLR4可抑制相关炎症因子的表达,提示TLR4可能通过诱导微炎症反应而参与高血压肾损害.【总页数】5页(P304-308)【作者】汤天凤;唐荣;蒋春明;夏阳阳;敖翔;周巧玲【作者单位】南京大学附属鼓楼医院肾内科,南京210008;中南大学湘雅医院肾内科,长沙410008;南京大学附属鼓楼医院肾内科,南京210008;南京大学附属鼓楼医院肾内科,南京210008;中南大学湘雅医院肾内科,长沙410008;中南大学湘雅医院肾内科,长沙410008【正文语种】中文【中图分类】R692.6【相关文献】1.血管紧张素Ⅱ1型受体表达对高糖诱导大鼠肾小管上皮细胞损伤的影响 [J], 查冬青;姚涛;高苹;吴小燕2.血管紧张素-(1-7)对血管紧张素II诱导的肾小管上皮细胞E-cadherin、α-SMA 表达的影响 [J], 袁波;贺红焰;王明勇;刘建3.胰高血糖素样肽-1类似物对高糖诱导下肾小管上皮细胞Toll样受体4、骨桥蛋白表达的影响 [J], 孔瑜;兰丽珍;张继敏4.RNA干扰技术沉默TRAF6对脂多糖诱导肾小管上皮细胞表型转化及炎症因子表达的影响 [J], 刘明龙;梁耀军;李侃;杨洋;马占军;吴蓉;王凤丽5.LncRNA NORAD对高糖诱导的人肾小管上皮细胞炎症因子和纤维化因子表达的影响及机制研究 [J], 卢迪;黎瑶;李婷;邱平因版权原因,仅展示原文概要,查看原文内容请购买。
福辛普利、氯沙坦对肾小管上皮细胞TLR4表达的影响汤天凤;周巧玲;朱俪俪;唐荣;敖翔【期刊名称】《中南大学学报(医学版)》【年(卷),期】2008(33)10【摘要】目的:探讨Toll样受体4(TLR4)在高血压肾损害中的作用机制以及福辛普利(fosinopril,Fos)、氯沙坦(Iosartan,Los)的肾脏保护机制.方法:将大鼠肾小管上皮细胞(NRK-52E)分5组:正常时照组、NRK-52E+Ang Ⅱ组、NRK-52E+AngⅡ+Fos(10-5mmol/L)组、NRK-52E+Ang Ⅱ+Los(10-5 mmol/L)组、NRK-52E+AngⅡ+Fos(10-5 mmol/L)+Los(10-5 mmol/L)组,培养24 h进行检测;同时建立稳定转染TLR4-specific RNAi质粒的NRK-52E细胞株.RT-PCR检测TLR4,IL-6,TNF-a mRNA表达;Western印迹检测TLR4蛋白表达;免疫细胞化学方法观察NF-KB核易位情况;酶联免疫吸附法(ELISA)检测上清IL-6及TNF-a水平.结果:Ang Ⅱ可上调TLR4,IL-6,TNF-a的表达水平,诱导NF-KB核易位活化(P<0.01);TLR4-specific RNA干扰后NRK-52E中TLR4水平显著下降(P<0.01),Ang Ⅱ刺激转染后的NRK-52E,其TLR4,IL-6,TNF-a水平较NRK-52E+Ang Ⅱ组显著下降(P<0.01),NF-KB核活化水平无明显变化(P>0.05).Fos或/和Los干预后可明显下调TLR4,IL-6,TNF-a表达,显著抑制NF-kB活化(P<0.01或P<0.05),联合用药与单药组无统计学差异(P>0.05).结论:TLR4在高血压肾损伤中可能有致炎作用.Fos和Los治疗高血压肾损害的作用机制之一,可能是通过下调TLR4基因及相关炎症介质的表达.【总页数】8页(P958-965)【作者】汤天凤;周巧玲;朱俪俪;唐荣;敖翔【作者单位】中南大学湘雅医院肾内科,长沙,410008;中南大学湘雅医院肾内科,长沙,410008;解放军163医院空勤科,长沙,410003;中南大学湘雅医院肾内科,长沙,410008;中南大学湘雅医院肾内科,长沙,410008【正文语种】中文【中图分类】R692【相关文献】1.青蒿琥酯对高糖诱导的肾小管上皮细胞TLR4、NF-κB表达及合成的影响 [J], 蒋姗姗;龙艳;苏珂;聂寒;杨帆;郑天鹏;莫如芬2.右美托咪啶对脂多糖致伤大鼠肾小管上皮细胞TLR4表达的影响﹡ [J], 雷安锋;段晶;陈燕;彭春晓;王秋月3.LPS对BALB/C小鼠肾小管上皮细胞TLR4和TNFα、IL-6表达的影响 [J], 朱晓玲;Li Ke;王永钧;杨亚珍;Wuding Zhou;Steven Sacks4.氯沙坦对高糖环境下人近端肾小管上皮细胞TGF-β1, PA1-1表达的影响 [J], 刘芳;石晓聪;陈雪;吴海波;郑海飞5.软脂酸对肾小管上皮细胞TLR4表达的影响 [J], 张鑫;刘晓玲;王兴兴;苏娟;王荣因版权原因,仅展示原文概要,查看原文内容请购买。
附子、黄连寒热药性对体外足细胞增殖及TRPV4、TRPC6蛋白表达的影响王竹;孙万森;李欣;李睿萍【摘要】目的研究附子、黄连寒热药性对体外足细胞增殖及TRPV4、TRPC6蛋白表达的影响. 方法 MTT法检测足细胞增殖,实验分为:空白对照组、附子组和黄连组.附子组和黄连组加入不同浓度药液进行干预,并测定吸光值(A),计算细胞抑制率.检测TRPV4、TRPC6蛋白试验中,实验分为:空白对照组、附子组和黄连组,其中附子组给予200 μg/ml药液、黄连组给予800 μg/ml药液,再加入含RPMI 1640培养基培养足细胞24h.用Western blot法检测附子、黄连对TRPV4、TRPC6蛋白表达影响. 结果 MTT的研究表明,黄连组中50,100,200,400,800 μg/ml药液均可抑制足细胞生长增殖,且与浓度成正相关.附子在相对较低的质量浓度范围内(< 200 μg/ml),促进足细胞生长增殖,超过一定实验浓度时(>200 μg/ml),抑制足细胞生长增殖.实验表明附子和黄连均可以上调TRPV4蛋白表达,与空白对照组相比差异有统计学意义(P<0.05).而在TR-PC6蛋白表达中,附子和黄连具有明显的下调TRPC6蛋白表达的作用,与空白对照组对比差异有统计学意义(P<0.05).结论附子、黄连这两种不同药性的中药均可以影响与肾脏病相关的蛋白TRPV4、TRPC6表达.%Objective To explore the effects of Fuzi and Huanglian on podocyte proliferation and the expression of TRPV4 and TRPC6 proteins invitro.Methods ①MTT method was used to detect the podocyte proliferation.The experiment was divided into blank control group,Fuzi group and Huanglian group.Different concentrations of Fuzi and Huanglian were used to treat the podocytes.The absorbance value(A) was measured,and the inhibi tion rate of cells was calculated.②The experimentwas divided into blank control group,Fuzi(200 μg/ml) group and Huanglian(800 μg/ml) group.The podocytes were given liquid medicine,and then cultured with RPMI 1640 medium for 24 h to detect the expression of TRPV4 and TRPC6 protein by Western blot.Results MTT results showed that 50,100,200,400 and 800 μg/ml Huanglian inhibited the growth and proliferation of podocyte in a concentration-dependent manner.The relatively low concentration of Fuzi (<200 μg/ml) pr omoted the growth and proliferation of podocytes,while the relatively high concentration(>200 μg/ml) inhibited the growth and proliferation of podocytes.The results showed that the expression of TRPV4 protein in Fuzi group and Huanglian group were significantly increased compared with blank control group (P < 0.05),while the expression of TRPC6 protein in Fuzi and Huanglian group were decreased (P < 0.05).Conclusion The traditional Chinese medicine Fuzi and Huanglian all have the influence on the expression of TRPV4 and TRPC6 in kidney disease.【期刊名称】《山西医科大学学报》【年(卷),期】2017(048)010【总页数】4页(P999-1002)【关键词】寒热药性;TRPV4;TRPC6【作者】王竹;孙万森;李欣;李睿萍【作者单位】西安交通大学第二附属医院中医科,西安710004;西安交通大学第二附属医院中医科,西安710004;西安交通大学第二附属医院中医科,西安710004;西安交通大学第二附属医院中医科,西安710004【正文语种】中文【中图分类】R363中药的四性,即药物寒、热、温、凉之性,反映药物作用于人体寒热变化的影响,是中药临床运用的核心。
补肾通络方对衰老大鼠血清T水平及Leydig细胞数目的影响杨光照;邓冬梅;庄天衢【期刊名称】《河南中医》【年(卷),期】2012(32)7【摘要】目的:观察补肾通络方对雄激素部分缺乏大鼠血清睾酮(T)水平及睾丸间质(Leydig)细胞数目的影响。
方法:采用自然衰老大鼠模型,补肾通络方低、中、高剂量组干预模型大鼠,放免法检测各组大鼠T、黄体生成素(LH)的变化;HE染色观察Leydig数目变化;免疫组化法观察Leydig细胞StAR阳性表达。
结果:补肾通络方干预后能提高自然衰模型大鼠T水平(P<0.05),对LH影响不明显(P>0.05)。
补肾通络方高、中剂量组能增加雄激素部分缺乏大鼠Leydig细胞数目(P<0.05)。
结论:补肾通络方能提高模型大鼠血清T水平,增加Leydig细胞数目是其作用机制之一。
【总页数】3页(P833-835)【关键词】补肾通络方;衰老;血清T水平;Leydig细胞;大鼠【作者】杨光照;邓冬梅;庄天衢【作者单位】苏州中医院;苏州相城中医院【正文语种】中文【中图分类】R285.5【相关文献】1.针灸联合温脾补肾方对急性溃疡性结肠炎大鼠血管活性肠肽、血清肿瘤坏死因子-α和白细胞介素水平的影响 [J], 赵凤玲;常玉洁;董雪莲;张晓蕾2.补肾通络疏肝方对衰老大鼠阴茎组织转化生长因子β基因表达的影响 [J], 胡海翔;骆斌;董静3.补肾通络疏肝方对衰老大鼠阴茎组织一氧化氮合成酶基因表达的影响 [J], 胡海翔;刘占民;蔡庆;王喆4.补肾益元方对运动性低血睾酮大鼠睾丸Leydig细胞睾酮合成酶的影响 [J], 王一蓉;周志宏;汤长发;魏霞;叶志文5.补肾通脉方对2型糖尿病大鼠血清细胞因子及相关激素水平的影响 [J], 曾凡鹏;陆付耳;杨明炜;王开富;徐丽君因版权原因,仅展示原文概要,查看原文内容请购买。
四妙散加味方对人肾小管上皮细胞UAT和URAT1基因表达的影响王华杰;张国华;邹震;范秀珍【期刊名称】《中国临床药理学与治疗学》【年(卷),期】2010()9【摘要】目的:观察低、高剂量四妙散加味方(SMSJWF)是否调节人肾小管上皮细胞(HK-2细胞)尿酸盐转运子(hUAT)mRNA和有机阴离子转运体(hURAT1)mRNA 的表达。
方法:根据培养液中所含四妙散加味方剂量的不同,将HK-2细胞分为:DMEM/F-12+6 mL/2 kg四妙散加味方低剂量组;DMEM/F-12+12 mL/2 kg 四妙散加味方高剂量组;DMEM/F-12+苯溴马隆4.8 mg/d阳性对照组;仅使用DMEM/F-12+空白血清对照组。
四组细胞分别培养48 h,采用实时荧光定量PCR 检测HK-2细胞中hUATmRNA、hURAT1 mRNA的相对表达量(2△△Ct法)。
结果:所有标本均能检测到hUAT mRNA和hURAT1 mRNA的表达。
低、高剂量组和阳性对照组hUAT mRNA表达水平均明显高于空白对照组(P<0.05);低剂量和阳性对照组的hURAT1 mRNA的表达明显低于空白对照组(P<0.05),而高剂量组hURAT1 mRNA的表达明显高于其他各组(P<0.05)。
结论:低、高剂量四妙散加味方可上调hUAT mRNA表达;低剂量四妙散加味方与苯溴马隆均可下调hURAT1mRNA的表达,这可能是其降尿酸的作用机制之一。
【总页数】6页(P972-977)【关键词】四妙散加味方;尿酸;肾小管;基因表达;转运蛋白【作者】王华杰;张国华;邹震;范秀珍【作者单位】邵阳市第一人民医院;湖南省中医研究院附属医院【正文语种】中文【中图分类】R311;R965.2【相关文献】1.四妙散对尿酸诱导人肾小管上皮细胞结缔组织生长因子及骨形成蛋白-7表达的影响 [J], 康洁;陈继承;张凌媛;丁珊珊2.Cyr61基因过表达对人肾小管上皮细胞表达细胞外基质成分的影响 [J], 沈学飞;梅长林;张岩;刘亚伟;戴兵;许涛;王文靖3.益气活血降浊方对TGF-β 1诱导人近端肾小管上皮细胞HGF、BMP-7表达的影响 [J], 冯博;刘文军;徐万忠;路晓光;刘瑞华4.狼疮方含药血清对人近端肾小管上皮细胞过氧化物酶体增殖物激活受体γ表达及CD40、RANTES的影响 [J], 张亚杰;阳晓;陈伟英;孔庆瑜;董秀清;余学清5.清热泄浊化瘀方对人肾小管上皮细胞UAT和URAT1基因表达的影响 [J], 王华杰;张国华;邹震;范秀珍因版权原因,仅展示原文概要,查看原文内容请购买。
TRPV4在小鼠脑血管内皮细胞中的表达及对血管张力调节作用研究胡文霞;姜婉;叶立;叶沛;朱金行;沈兵;杜鹃【摘要】目的比较瞬时受体电位(TRP)通道家族中香草素受体亚家族(TRPV)和经典瞬时感受器电位亚家族(TRPC)在小鼠脑、心和肝微血管内皮细胞表达差异,寻找脑微血管中具有特异性高表达亚型,阐明其对脑动脉血管舒张的调节作用.方法挖掘GEO数据库中芯片表达谱数据,分析各个通道亚型差异表达情况,再采用免疫组化检测TRPV4、TRPP2和TRPC1在脑基底动脉中表达情况,血管张力实验检测TRPV4通道在调节小鼠脑基底动脉舒张中的效应.结果通过分析表达谱数据显示,在小鼠脑、心和肝三种微血管内皮中,TRPV4通道在脑微血管内皮细胞显著高表达,而TRPC3通道显著低表达.TRPV4、TRPP2和TRPC1在脑基底动脉内皮和平滑肌层也显著表达.TRPV4通道激动剂GSK1016790A能浓度依赖地舒张苯肾上腺素引起的脑基底动脉收缩,而且其舒张效应可以被TRPV4通道阻断剂HC067047所阻断.结论与心和肝血管内皮细胞相比,TR-PV4通道在小鼠脑血管内皮细胞中显著高表达,可能在调节脑血管内皮功能中具有重要意义.%Objective To compare the expression difference of transient receptor potential subfamilies including transient receptor potential vanilloid(TRPV) and transient receptor potential canonical (TRPC) channels in mouse microvascular endothelial cells from brain,heart and liver,and find a specific high expression subtypes in brain microvessels endothelial cells,to clarify its regulation function in cerebral vasodilatation.Methods The array data for the expression profiling from GEO Databases was analyzed to clarify the expression profiles of all TRPV and TR-PC channel subtypes.Then immunohistochemistry was used toidentify the expression profile of TRPV4,TRPP2 and TRPC1 in the cerebral basilar artery,and vessel tension measurement was used to investigate the regulatory effect of TRPV4 channel having high expression level on the cerebral basilar artery relaxation.Results The results showed that in three types of endothelial cells from brain,heart and liver,the expression level of TRtV4 channel was highest but the expression level of TRPC3 was lowest in brain microvascular endothelial cells.TRPV4,TRPP2 and TRPC1 were expressed in the endothelial and smooth muscle layers of the cerebral basilar artery as well.GSK1016790A,an agonist of TRPV4,concentration-dependently relaxed the cerebral basilar artery which was preconstructed by phenylephrine.Additionally,GSK1016790A-induced vessel relaxation was inhibited by HC067047,an antagonist of TRPV4.Conclusion Compared with the microvascular endothelial cells from heart and liver,the brain microvascular endothelial cells show significantly high expression in TRPV4 channel.Therefore,TRPV4 may have a critical role in the function of brain microvascular endothelial cells.【期刊名称】《安徽医科大学学报》【年(卷),期】2017(052)010【总页数】5页(P1433-1437)【关键词】瞬时受体电位离子通道;内皮细胞;TRPV4;脑血管【作者】胡文霞;姜婉;叶立;叶沛;朱金行;沈兵;杜鹃【作者单位】安徽医科大学六安市人民医院神经内科,六安237005;安徽医科大学基础医学院生理学教研室,合肥230032;安徽医科大学基础医学院生理学教研室,合肥230032;安徽医科大学基础医学院生理学教研室,合肥230032;安徽医科大学六安市人民医院神经内科,六安237005;安徽医科大学基础医学院生理学教研室,合肥230032;安徽医科大学基础医学院生理学教研室,合肥230032;安徽医科大学基础医学院生理学教研室,合肥230032【正文语种】中文【中图分类】Q412瞬时受体电位(transient receptor potential, TRP)通道属于非选择性阳离子通道家族,由于大多数TRP通道都通透钙离子,所以其在介导细胞钙信号中具有重要作用[1]。
•实验研究•健脾益气通络中药对糖尿病肾病大鼠肾组织病理变化的影响!李琼锋,韦衮政,徐蕾,王宫琳(云南省中医医院,云南昆明650021)摘要*目的观察健脾益气通络中药对糖尿病肾病大鼠肾组织病理变化的影响。
方法将实验用的60只健康雄性S D大鼠,随机法分为6组:左肾切除组、模型组、洛汀新组、中药低、中、高剂量组,每组收入10只,除肾切除组外,其余五组均构建糖尿病肾病大鼠模型,并对中药低、中、高剂量组进行对应剂量的健脾益气通络中药灌胃,洛汀新组进行洛汀新灌胃,对左肾切除组、模型组进行等量生理盐水灌胃,灌胃6周后,检测各组大鼠24h的尿蛋白(U p<)定量、肌酐(Scr)、尿素氮(B U N)。
取肾脏开展H E染色行电镜观察以及透射电镜观察肾皮质组织病理改变。
结果模型组大鼠与肾切组大鼠相比,光镜及电镜下已出现糖尿病肾病病理改变,各治疗组肾皮质病理改变较轻;洛汀新组、中药低、中、高剂量组与模型组比较,24h尿蛋白(U p<)定量显著下降(E<0.01);各治疗组与模型组比较,尿素氮(B U N)、肌酐(S c)明显降低(E<0. 05)。
结论健脾益气通络中药可改善糖尿病肾病大鼠的肾功能,并可减轻肾脏的病理损害。
关键词:健脾益气通络中药;糖尿病肾病;大鼠;病理形态学中图分类号:R285.5 文献标志码:A文章编号*1007 -2349(2019)01 -0063 -03@fect of Jianpi Yiqi Tongluo Recijje on Pathological Changes o f Renal Tissue in Rats 7tli Diabetic N ephropa LI Q i o n g -f e n g,W E I G u n -z h e n g,X U L e i,W A N G G o n g -lin(Yunnan Provincial Hospital of Traditional Chinese Medicine,Kunming650021,China)3Abstract】Objective:To observe the effect of Jianpi Yiqi Tongluo Recipe on the pathological change betic nephropathy.M e t h o d s: 60 h ealthy male SDrats were randomly divided into6 groups:left nephrectomy group,model group,lotensingroup,l o w,m e d i u m and high dose T C M groups,10 in each group.Except the nephrectomy group,the other five groups were establishedas rat models of diabetic nephropathy.The l o w,m e d i u m and high dose T C M groups were given in dose of Jianpi Yiqi Tongluo,and lotensin group was were intragastrically administrated with lotensin.The rats in and the model group were intragastrically administered with normal saline.After 6 weeks of gavage,the urine protein (U p r o),creatinine(Scr)and urea nitrogen (B U N)were measured in each group for24 hours.The kidneys were subjected to H E staining by electron microscopy and transmission electron microscopy to observe the patliological changes of renal cortex tissue.Results:C o tomized rats,there was the pathological changes of diabetic nephropathy in the model group under electron microscopy and transmission electron microscopy and there was lighter pathological changes of renal cortex in each treatment group.C o m pared with the model group,the24h urine protein(U p r o)ration of the lotensin group and the l o w,m e d i u m and high dose T C M groups was significantly decreased(P <0. 01). C o m pared with the model group,the urea nitrogen (B U N)and creatinine(Scr)in each treatment group were significantly lowerthan those in the model group (P< 0. 05 ). Conclusion:Jianpi Yiqi Tongluo Recipe can improve the renal function of rats with diabetic nephropathy and reduce the pathological damage of the kidney.【K e y w o r d s】Jianpi Yiqi Tongluo Recipe,diabetic nephropathy,rat,pathological morphology糖尿病肾病(3N)属于目前世界多发病和难治病,治疗手段有限。
祛风通络方对单侧输尿管梗阻大鼠肺肾组织AQP4表达的影响∗刘艳1,谢君2,王竹1,李睿萍1,孙万森1△1西安交通大学第二附属医院,陕西西安710004;2西安市胸科医院[摘要]目的:观察祛风通络方对单侧输尿管梗阻大鼠肺肾组织水通道蛋白(aquaporin-4,AQP4)表达的影响,探讨机体水液代谢功能的分子基础。
方法:建立单侧输尿管梗阻大鼠模型,给予大、小剂量祛风通络方干预20天,采用免疫组织化学法、RT-PCR及Western blot方法检测AQP4在肺、肾组织的分布及表达情况。
结果:模型组大鼠AQP4表达降低,祛风通络方干预组AQP4表达均升高,大剂量组升高较明显。
结论:AQP4是肺、肾参与水液代谢的分子基础之一,祛风通络方有上调AQP4的作用。
[关键词]输尿管梗阻;AQP4;水液代谢;祛风通络方;大鼠[中图分类号]R692[文献标识码]A[文章编号]2096-9600(2021)03-0043-04Effects of Qufeng Tongluo Prescription on the Expressions of AQP4in Lung and Kidney of the Rats Suffering Unilateral Ureteral ObstructionLIU Yan1,XIE Jun2,WANG Zhu1,LI Ruiping1,SUN Wansen1△1The Second Affiliated Hospital of Xi′an Jiaotong University,Xi′an710004China;2Xi′an Chest HospitalAbstract Objective:To observe the effects of Qufeng Tongluo prescription on the expressions of AQP4in lung and kidney of the rats with unilateral ureteral obstruction(UUO),and explore the molecular basis of water metabolism.Methods:After establishing rat model with UUO,they were intervened with large and small dose of Qufeng Tongluo prescription for20days,immunohistochemical method,RT-PCR and Western blot method were adopted to detect the distribution and the expressions of AQP4in lung and kidney.Results:The expressions of AQP4in the model group reduced,while these raised in the intervention group of Qufeng Tongluo prescription, the rise in large dose group was more notable.Conclusion:AQP4is one of the molecular basis of lung and kidney involved in water metabolism,and Qufeng Tongluo prescription could up-regulate AQP4.Keywords ureteral obstruction;AQP4;water metabolism;Qufeng Tongluo prescription;rats汪昂《医方集解》云:“肺为水之上源,肾为水之下源”,水通道蛋白(aquaporin-4,AQP4)与机体水液代谢密切相关。
不同药性祛风通络方干预下UUO 大鼠肾小管上皮细胞TRPV4变化规律研究**本课题为国家自然科学基金资助项目(No.81072974)①西安交通大学第二附属医院(西安710004)②陕西省西安市儿童医院(西安710043)△通讯作者马巧亚①李睿萍①孙万森①△铁晓玲②王竹①刘艳①〔摘要〕目的:通过观察祛风通络方及寒热加味方对UUO 模型大鼠肾组织TRPV4表达及功能状态的影响,探索不同药性祛风通络方对TRPV4的调节作用及其对肾间质纤维化的防治机理。
方法:挑选SD 雄性成年大鼠80只,随机分为空白对照组和模型组,单侧输尿管结扎法建立UUO 模型,分为模型对照组、温热大剂量组、温热小剂量组、寒凉大剂量组、寒凉小剂量组、祛风大剂量组、祛风小剂量组,进行药物干预。
术后第21天将大鼠全部处死,取术侧肾脏组织,一半用作病理学染色,一半用于分子生物学检测。
RT -PCR和Western 法检测肾脏组织TRPV4、α-SMA 、vimentin mRNA 含量变化及蛋白表达情况。
结果:(1)造模成功后21d ,模型组及干预组大鼠术侧肾脏体积较对侧增大,剖面肾实质不同程度萎缩,肾盂扩张呈囊状,模型组最为明显。
(2)干预各组病理情况均不同程度改善,其中温热小剂量组改善最为明显。
(3)温热大剂量组TRPV4mRNA 含量明显增加(P <0.01),其余各组均表现不同程度下降。
(4)治疗各组α-SMA mRNA 含量均表现不同程度下降(P <0.05),以温热小剂量组下降最为显著(P <0.01),各组Vimentin 的表达表现出与α-SMA 同样的变化趋势。
结论:(1)TRPV4可能通过基因或蛋白水平的上调(下调)感受药性之寒热。
(2)祛风通络方及加味方可能延缓肾间质纤维化的进展,且温热小剂量治疗效果最好。
〔关键词〕祛风通络方寒热药性单侧输尿管梗阻(UUO )瞬时感受电位香草酸受体4(TRPV4)Modulation of Transient Receptor Potential Vanilloid 4in Rats Kidneyby Different Nature Qufeng Tongluo Recipes RespectivelyMA Qiaoya ,LI Ruiping ,SUN Wansen ,et alSecond Affiliated Hospital of Xi ’an Jiaotong University ,Xi ’an (710004)ABSTRACTObjective :We observed the expression of TRPV4in renal tubular epithelial cells ,alpha -smooth muscle actin(α-SMA )and Vimentin in renal interstitial fibrosis induced by unilateral ureteral obstruction in rats.Adult male Sprague -Dawley rats (180 220g body weight )were subjected to unilateral ureteral obstruction by ligation of the left ureter.Methods :Eight groups of rats comprising 10animals which (total =80)were included as follows :sham -operated group ;non -treated UUO group ;large -dose warm ;low -dose warm group ;large -dose cold group ;low -dose cold group ;large -dose Qufeng group ;low -dose Qufeng group.Rats in Sham and UUO groups were administrated with distilled water.All of the rats were killed on day 21after UUO or Sham operation.The left kidney were harvested for various biochemical and morphological study.Masson ’s trichrome staining was used tostudy the dynamic histological changes.TRPV4,α-SMA ,Vimentin mRNA and proteins were evaluated by real time RT -RCRand western blotting.Results :(1)On day of 21,the obstructed and contralateral kidneys were weighed and a ratio was calculated.In UUO group ,this ratio was significantly higher than in sham -operated animals.Meanwhile ,renal parenchymal thickness was decreased com-pared with the contralateral kidney ,dilated calyces and pelviectasis were mark following UUO.(2)Compared with sham -operated ,an obvious tubulointerstitial injury was characterized with UUO by tubular dilatation and atrophy ,fibroblasts ,myofibroblasts ,macro-phages ,and collagen ,which were reduced inconsistently in all treatment groups.This amelioration was conspicuouous in low -dose warm group.(3)Different nature Qufeng tongluo recipes could down -regulate the high expression expression of mRNA of TRPV4in-duced by UUO except large -dose warm recipe (P <0.01).(4)Different nature Qufeng tongluo recipes therapy decreased its levels of mRNA and protein (P <0.05),and it represented the most significant down -regulation in low -dose warm group (P <0.01).In general ,the change of Vimetin kept consistence with α-SMA.Conclusion :(1)TRPV4may be up -regulated (down -regulated )by gene or protein levels to sense the coldness of the drug.(2)Qufeng Tongluo Recipe and Jiawei Recipe may delay the progression of re-nal interstitial fibrosis ,and the best effect of warming small doses.KEY WORDS Qufeng tongluo recipes Cold and heat properties Unilateral ureteral obstruction Transient receptor poten-tial vanilloid4中医药在慢性肾脏病的治疗中有着独特的优势,单味中药或中药提取物,或辨证组方在动物实验、体外研究及临床观察中均显示良好的抗纤维化作用和临床治疗效果。
而中医对慢性肾病证候的研究认为,水湿停聚是各种慢性肾脏疾病最基本的病理表现,而且在治疗疾病过程中强调“整体观念”,注重局部与全身的协调和阴阳平衡。
因此,深入研究中药对机体微环境的整体调节作用,进一步阐释中药的治疗机理尤为重要。
本研究通过单侧输尿管结扎术建立经典肾间质纤维化模型(UUO模型),观察UUO病理过程中肾小管上皮细胞上TRPV4、肾间质纤维化相关蛋白α平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)和波形蛋白(Vimentin)的变化,探索不同药性祛风通络方对TRPV4的调节作用及其对肾间质纤维化的防治机理,通过寒热药性的瞬时受体电位通道假说初步揭示中药可能的性-效本质。
材料与方法1实验药物制备及剂量确定祛风通络方(袪风小剂量组):生黄芪30g,生地30g,海风藤21g,乌梢蛇21g,防风21g。
祛风大剂量组:各药物用量均加倍。
温热祛风通络方(温热方)大剂量组:袪风方+附子30g;温热祛风通络方(温热方)小剂量组:袪风方+附子10g;寒凉祛风通络方(寒凉方)大剂量组:袪风方+黄连30g;寒凉祛风通络方(寒凉方)小剂量组:袪风方+黄连10g。
2实验动物与分组SD雄性成年大鼠80只,体重180 220g,由西安交通大学医学院动物实验中心提供,相同条件下喂养1周后随机分为两组:空白对照术组(A组,n=10)、单侧输尿管结扎(UUO)组(n=70),其中UUO组又根据干预的不同随机分为7组,即模型对照组(B组,n=10);温热大剂量组(C组,n=10),温热小剂量组(D组,n=10),寒凉大剂量组(E组,n=10),寒凉小剂量组(F组,n=10),祛风大剂量组(G组,n=10),祛风小剂量组(H组,n=10)。
3大鼠UUO模型的建立具体方法如下:80只大鼠随机分为8组,相同条件下适应性喂养1周,手术当日晨大鼠禁食、禁水,术前10%水合氯醛(2ml/kg)腹腔注射麻醉。
麻醉满意后,大鼠仰卧位固定于手术台上,腹部脱毛、消毒、铺巾,于剑突下2cm行腹正中直切口,长约3cm,逐层切开皮肤、皮下组织及腹膜。