摘要
目的研究桑枝多糖基于YAP/TAZ信号通路调控db/db小鼠肾纤维化作用的影响。方法将40只24周龄自发型2型糖尿病db/db小鼠随机分为模型组、缬沙坦组(20mg/kg)、桑枝多糖低剂量组(600mg/kg)、桑枝多糖高剂量组(1200mg/kg),每组10只,日常进行高脂饮食喂养,连续灌胃相应药物60天,每天1次。另将10只db/m小鼠为空白组,日常进行普通饲料喂养,模型组与空白组小鼠灌胃等体积生理盐水60天。给药第60天采集小鼠尿液,测量尿量及终点法测24h尿白蛋白(UP)含量。全自动生化分析仪测定小鼠血清肌酐(Scr)、尿素氮(BUN)含量。末次给药后,小鼠禁食不禁水12h,测其空腹血糖(FBG)。HE染色观察小鼠肾组织病理学变化。采用ELISA法检测肾组织纤维化指标TGF-β1、纤连蛋白(FN)、结缔组织生长因子(CTGF)、胶原Ⅰ型(CollagenⅠ)含量及炎症因子肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)、趋化因子单核细胞趋化蛋白-1(MCP-1)含量。Western blot法检测肾组织Yes相关蛋白(YAP)、TAZ蛋白表达。结果与模型组比较,桑枝多糖低、高剂量组小鼠Scr[(0.39±0.09)mg/dL、(0.32±0.07)mg/dL比(0.62±012)mg/dL]、BUN[(25.75±4.75)mg/dL、(20.62±3.68)mg/dL比(39.18±4.63)mg/dL]、UP[(3.45±0.42)mg/24h、(2.76±0.47)mg/24h比(6.38±2.03)mg/24h]含量显著降低(P均<0.05);纤维化指标TGF-β1[(3.76±0.52)ng/mL、(2.53±0.38)ng/mL比(4.98±1.17)ng/mL]、FN[(5.29±1.13)ng/mL、(4.47±1.02)ng/mL比(6.13±1.07)ng/mL]、CTGF[(14.52±1.22)ng/mL、(11.24±0.87)ng/mL比(17.52±1.26)ng/mL]、CollagenⅠ[(10.93±0.85)ng/mL、(8.14±0.76)ng/mL比(14.81±1.34)ng/mL]及炎症因子TNF-α[(87.25±7.81)ng/L、(78.66±6.36)ng/L比(134.82±8.90)ng/L]、IL-6[(34.72±3.35)ng/L、(21.16±2.51)ng/L比(58.65±5.71)ng/mL]、MCP-1[(0.38±0.04)ng/L、(0.29±0.06)ng/L比(0.51±0.05)ng/L]含量明显减少(P均<0.05),YAP[(0.48±0.06)、(0.31±0.05)比(0.53±0.11)]、TAZ[(0.31±0.04)、(0.21±0.03)比(0.37±0.05)]蛋白表达显著降低(P均<0.05)。结论桑枝多糖对db/db小鼠糖尿病肾病的肾纤维化有一定的保护作用,其作用机制可能与调节YAP/TAZ信号通路,降低肾脏组织纤维化因子形成及减少炎症因子含量有关。
Objective To investigate the effect of Ramulus Mori polysaccharide(RMP)on renal fibrosis in db/db mice through Yes-associated protein(YAP)//Tafazzin(TAZ)signal pathway.Methods Twenty-four weeks'old spontaneous db/db type 2 diabetic mice were randomly divided into four groups:model,valsartan(20mg/kg),low dose(600mg/kg)and high dose(1200mg/kg)of RMP groups(n=10 per group).Mice were fed with high-fat diet and intragastrically administered with the relevant drug once a day for 60 days.Additional 10 mice were selected as a control group,fed with standard rodent food and given equally allocated volume of saline for 60 days.On the 60th day of experiment,24-hour urine samples were collected,and urine volume and 24 h urine albumin(UP)were analyzed;serum creatinine(Scr)and urea nitrogen(BUN)were determined by automatic biochemical analyzer;fasting blood glucose(FBG)was determined after 12 h starving.Additionally,the renal tissue samples were collected and stained with HE.Serum transforming growth factor-β1(TGF-β1),fibronectin(FN),connective tissue growth factor(CTGF),collagen type I(Collagen I),tumor necrosis factor(TNF-α),interleukin-6(IL-6)and monocyte xhemotactic protein-1(MCP-1)were measured by ELISA for assessment of renal fibrosis.Protein expressions of YAP and TAZ in renal tissues were analyzed by Western-blot.Results Compared to the model group,the MRP low and high dose groups(model vs.MRP-low vs.MRP-high)showed significantly lower levels of Scr(0.39±0.09 vs.0.32±0.07 vs.0.62±0.12mg/dL)and BUN(25.75±4.75 vs.20.62±3.68 vs.39.18±4.63mg/dL)(all P<0.05),and lower serum levels of TGF-β1(3.76±0.52 vs.4.98±1.17 vs.2.53±0.38ng/mL),FN(5.29±1.13 vs.4.47±1.02 vs.6.13±1.07ng/mL),CTGF(14.52±1.22 vs.11.24±0.87 vs.17.52±1.26ng/mL),CollagenⅠ(10.93±0.85 vs.8.14±0.76 vs.14.81±1.34ng/mL),TNF-α(87.25±7.81 vs.78.66±6.36 vs.134.83±8.90ng/L),IL-6(34.72±3.35 vs.21.16±2.51 vs.58.65±5.71ng/L)and MCP-1(0.38±0.04 vs.0.29±0.06 vs.0.51±0.05ng/L)(all P<0.05).Compared to the model group,expressions of YAP and TAZ were both markedly downregulated(P<0.05)in either MRP group.Conclusion MRP could attenuate the progression of renal fibrosis morphologically.The protective effect of MRP on renal fibrosis in db/db mice might be involved in regulating the YAP/TAZ signal pathway.
作者
余文胜
黄飞
郑妮
YU Wen-sheng;HUANG Fei;ZHENG Ni(Affiliated Hangzhou First People's Hospital,Zhejiang University School of Medicine,Hangzhou,Zhejiang,310006;Hangzhou Obstetrics and Gynecology Hospital,Hangzhou,Zhejiang,310008;Southern University of Science and Technology Hospital,Guangdong Shenzhen,518000)
出处
《浙江中西医结合杂志》
2021年第7期603-608,共6页
Zhejiang Journal of Integrated Traditional Chinese and Western Medicine
基金
国家自然科学青年基金(No.81500633)。
