摘要
目的 基于Ras同源基因家族成员A(RhoA)/Rho相关卷曲螺旋形成蛋白激酶(ROCK)信号通路探讨茯苓酸(PA)对慢性肾衰竭(CRF)大鼠肾功能、纤维化的影响及潜在机制。方法 以雄性SD大鼠为对象,以5/6肾切除术建立CRF大鼠模型,将造模成功的大鼠分为模型组,PA低、中、高剂量组(5、10、20 mg/kg PA)和PA高剂量+ROCK通路激活剂溶血磷脂酸(LPA)组(20 mg/kg PA+1 mg/kg LPA),每组15只;另取15只大鼠仅切口暴露肾脏但不切除,作为假手术组。各药物组大鼠灌胃和/或尾静脉注射相应药液,每天1次,连续12周。实验期间,观察各组大鼠一般情况;末次给药后,检测各组大鼠的血清肾功能指标(血尿素氮、血清肌酐、尿酸)水平,观察其肾组织病理学改变,并进行肾小管损伤评分和肾纤维化区域面积量化,检测其肾组织中氧化应激指标[丙二醛(MDA)、超氧化物歧化酶(SOD)]、炎症因子(肿瘤坏死因子α、白细胞介素1β、白细胞介素6)水平,结缔组织生长因子(CTGF)、Ⅰ型胶原(collagenⅠ)阳性表达率,以及通路相关蛋白(RhoA、ROCK1)、纤维化相关蛋白(转化生长因子β1、裸角质层同源物2、α-平滑肌肌动蛋白)表达水平。结果 与假手术组比较,模型组大鼠饮食减少,体型瘦小,精神萎靡且反应迟钝,肾小球减少、萎缩,肾小管扩张且结构杂乱,间质可见大量炎症细胞浸润;其血清肾功能指标水平,肾小管损伤评分、肾纤维化面积占比、肾组织MDA和炎症因子水平、CTGF和collagenⅠ阳性表达率、通路相关蛋白和纤维化相关蛋白的表达水平均显著升高,SOD水平显著降低(P<0.05)。与模型组比较,PA各剂量组大鼠的一般情况和肾组织病理损伤均有不同程度好转,上述各定量指标均显著改善,且呈剂量依赖性(P<0.05)。LPA可显著逆转PA对上述指标的改善作用(P<0.05)。结论 PA能改善CRF大鼠肾功能并减轻肾组织纤维化,上述作用可能与抑制RhoA/ROCK信号通路的激活有关。
OBJECTIVE To investigate the effect of pachymic acid(PA) on renal function and fibrosis in chronic renal failure(CRF) rats and its potential mechanism based on the Ras homolog gene family member A(RhoA)/Rho-associated coiled-coil forming protein kinase(ROCK) signaling pathway.METHODS Using male SD rats as subjects,the CRF model was established by 5/6 nephrectomy;the successfully modeled rats were divided into model group,PA low-dose,medium-dose and high-dose groups(5,10,20 mg/kg PA),high-dose PA+ROCK pathway activator lysophosphatidic acid(LPA) group(20 mg/kg PA+1 mg/kg LPA),with 15 rats in each group.Another 15 rats were selected as the sham operation group with only the kidney exposed but not excised.The rats in each drug group were gavaged and/or injected with the corresponding liquid via the caudal vein,once a day,for 12 consecutive weeks.During the experiment,the general condition of rats was observed in each group.After the last administration,the serum renal function indexes(blood urea nitrogen,serum creatinine,uric acid) of rats in each group were detected,the renal histopathological changes were observed;the renal tubule injury score and the area of renal fibrosis were quantified.The levels of oxidative stress indexes [malondialdehyde(MDA),superoxide dismutase(SOD)] and inflammatory factors(tumor necrosis factor-α,interleukin-1β,interleukin-6),the positive expression rates of connective tissue growth factor(CTGF) and collagen Ⅰ were detected as well as the expression levels of pathway-related proteins(RhoA,ROCK1) and fibrosisrelated proteins(transforming growth factor-β1,bare corneum homologs 2,α-smooth muscle actin) were determined.RESULTS Compared with the sham operation group,the rats in model group had reduced diet,smaller body size,listless spirit and sluggish response,reduced and atrophied glomeruli,dilated renal tubules with chaotic structure,and a large number of inflammatory cells infiltrated interstitium;the serum levels of renal function indexes,renal tubule injury score,renal fibrosis area proportion,the levels of MDA and inflammatory factors,the positive expression rates of CTGF and collagen Ⅰ,and the expression levels of pathway-related proteins and fibrosis-related proteins in renal tissues were significantly increased,while SOD level was significantly decreased(P<0.05).Compared with the model group,the general condition and pathological injuries of kidney tissue of rats in PA groups were improved to varying degrees,and the above quantitative indexes were significantly improved in a dose-dependent manner(P<0.05).LPA could significantly reverse the improvement effect of PA on the above indicators(P<0.05).CONCLUSIONS PA can improve renal function and alleviate renal fibrosis in CRF rats,which may be related to inhibiting the activation of Rho A/ROCK signaling pathway.
作者
彭斌
冯雪
冯立
熊玮
胡溪
朱双益
肖阳
陈芳
高智
PENG Bin;FENG Xue;FENG Li;XIONG Wei;HU Xi;ZHU Shuangyi;XIAO Yang;CHEN Fang;GAO Zhi(Dept.of Nephrology,Wuhan Hospital of Traditional Chinese Medicine,Wuhan 430049,China)
出处
《中国药房》
CAS
北大核心
2024年第12期1489-1494,共6页
China Pharmacy
基金
武汉市医学科研项目(No.WZ21C25)。
