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21 d,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TRPV4 mRNA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。
Objective: We observed the expression of TRPV4 in 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 ~ 220 g body weight) were subjected to unilateral ureteral obstruction by ligation of the left ureter. Methods: Eight groups of rats comprising 10 animals 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 21 after 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-RCR and 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 compared 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,macrophages,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 TRPV4 induced 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 consis
作者
马巧亚
李睿萍
孙万森
铁晓玲
王竹
刘艳
MA Qiaoya;LI Ruiping;SUN Wansen(Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710004)
出处
《中国中西医结合肾病杂志》
2019年第1期15-19,共5页
Chinese Journal of Integrated Traditional and Western Nephrology
基金
国家自然科学基金资助项目(No.81072974)
