基于SphK1/S1PR2通路探讨肾必宁颗粒对IgA肾病大鼠肾组织的保护作用
Protective effect of Shenbining Granule on renal tissue of IgA nephropathy rats based upon SphK1/S1PR2 pathway
姜浩然 1宋纯东 1段凤阳 2王宁丽 2郭婷 2张博 2张霞 2丁樱 1任献青 1翟文生1
作者信息
- 1. 河南中医药大学第一附属医院儿科医院,河南郑州 450000;河南中医药大学儿科医学院,河南郑州 450046
- 2. 河南中医药大学第一附属医院儿科医院,河南郑州 450000
- 折叠
摘要
目的:探讨肾必宁颗粒通过鞘氨醇激酶1(sphingosine kinase 1,SphK1)/1-磷酸鞘氨醇受体2(sphingosine 1 phosphate receptor 2,S1PR2)通路改善IgA肾病(IgA nephropathy,IgAN)大鼠肾损伤的作用及机制.方法:将54只健康雄性SD大鼠按照完全随机分组法分为空白组(n=10)、模型组(n=10)、泼尼松组(n=10)、肾必宁低剂量组(n=12)和肾必宁高剂量组(n=12).除空白组外,其余4组建立IgAN模型,第9周起治疗组灌胃给药,7周后生化检测血肌酐(SCr)、尿素氮(BUN)、谷丙转氨酶(ALT)、血清白蛋白(ALB)、尿红细胞计数及24 h尿蛋白定量(24 h-UTP);苏木素-伊红(HE)染色观察各组大鼠肾组织病理学变化;免疫荧光观察IgA沉积;蛋白免疫印迹(Western blot)法检测SphK1、S1PR2蛋白表达;实时荧光定量PCR法(RT-PCR)检测SphK1、S1PR2 mRNA表达.结果:与空白组比较,模型组大鼠尿红细胞计数、24 h-UTP、SCr、BUN水平均显著升高(P＜0.01或P＜0.05),ALB水平显著降低(P＜0.01),ALT水平有所升高,但差异无统计学意义(P＞0.05),有明显的肾脏病理损伤,免疫荧光可见大量IgA荧光沉积,肾组织SphK1、S1PR2蛋白及mRNA表达水平显著升高(P＜0.01);与模型组比较,泼尼松组、肾低组、肾高组大鼠尿红细胞数、24 h-UTP、BUN水平均明显降低(P＜0.01),ALB水平显著升高(P＜0.01),各组ALT水平有所下降,但差异无统计学意义(P＞0.05),肾低组SCr水平显著降低(P＜0.05),肾脏病理有不同程度改善,IgA荧光沉积可见不同程度减少,肾组织SphK1、S1PR2蛋白及mRNA表达水平明显下降(P＜0.01).结论:肾必宁颗粒可能通过抑制SphK1/S1PR2通路来降低IgAN大鼠尿蛋白,减轻血尿并改善肾功能,从而起到保护肾脏、防治肾损害的作用.
Abstract
OBJECTIVE To explore the effect and mechanism of Shenbining Granule(SG)on renal injury in rats with IgA nephropathy(IgAN)through sphingosine kinase 1(SphK1)/sphingosine 1 phosphate receptor 2(S1PR2)pathway.METHODS A total of 54 healthy male Sprague-Dawley rats were randomized into 5 groups of blank(n=10),model(n=10),prednisone(n=10),low-dose SG(n=12)and high-dose SG(n=12).Except for blank group,IgAN model was established in the other four groups.Treatment groups received intragastric dosing from Week 9.After 7 weeks,serum creatinine(SCr),blood urea nitrogen(BUN),alanine aminotransferase(ALT),serum albumin(ALB),urinary erythrocyte count and 24-hour urinary protein quantity(24 h-UTP)were measured.Renal pathological changes were observed by hematoxylin-eosin(HE)stain and IgA deposition was detected by immunofluorescence.The protein expressions of SphK1 and S1PR2 were examined by Western blot and the expressions of SphK1 and S1PR2 mRNA by real-time fluorescent quantitative polymerase chain reaction(RT-PCR).RESULTS As compared with blank group,urinary erythrocyte count,the levels of 24 h-UTP,SCr and BUN spiked signifi-cantly in model group(P＜0.01 or P＜0.05).The level of ALB dropped obviously(P＜0.01)while the level of ALT rose.However,no significant difference existed between model and blank groups(P＞0.05).There was obvious renal pathological injury.Massive IgA deposition could be detected by immunofluorescence.The expression levels of SphK1,S1PR2 protein and mRNA jumped sharply in renal tissue(P＜0.01).As compared with model group,urinary erythrocyte count,the levels of 24 h-UTP and BUN declined obviously in prednisone,low-dose SG and high-dose SG groups(P＜0.01),ALB level became mark-edly elevated(P＜0.01)while ALT level declined.The differences were not statistically significant(P＞0.05).SCr level declined markedly in low-dose SG group(P＜0.05).Renal pathology improved and IgA fluorescent deposition diminished.The expressions of SphK1,S1PR2 protein and mRNA dropped significantly in renal tissue(P＜0.01).CONCLUSION SG may lower urinary protein,lessen hematuria and improve renal function in IgAN rats through an inhibition of SphK1/S1PR2 pathway.Thus it protects kidney and prevents renal damage.
关键词
肾必宁颗粒/IgA肾病/鞘氨醇激酶1/1-磷酸鞘氨醇受体2信号通路/肾损伤Key words
Shenbining Granule/IgA nephropathy/sphingosine kinase 1/sphingosine 1 phosphate receptor 2 pathway/renal injury引用本文复制引用
基金项目
国家自然科学基金面上项目(82074493)
河南省中医药学科领军人才项目(豫卫中医函20218号)
河南省卫生健康委国家中医临床研究基地科研专项(2022JDZX003)
河南省中医药科学研究重大专项(2023ZYZD02)
出版年
2024
NETL
NSTL
万方数据