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

作　　者：姜浩然 宋纯东[1,2] 段凤阳[1] 王宁丽 郭婷[1] 张博[1] 张霞[1] 丁樱[1,2] 任献青[1,2] 翟文生[1,2] JIANG Haoran;SONG Chundong;DUAN Fengyang;WANG Ningli;GUO Ting;ZHANG Bo;ZHANG Xia;DING Ying;REN Xianqing;ZHAI Wensheng(Pediatric Hospital,First Affiliated Hospi-tal,Henan University of Chinese Medicine,Henan Zhengzhou 450000,China;College of Pediatrics,Henan University of Chinese Medicine,Henan Zhengzhou 450046,China)

机构地区：[1]河南中医药大学第一附属医院儿科医院,河南郑州450000 [2]河南中医药大学儿科医学院,河南郑州450046

出　　处：《中国医院药学杂志》2024年第11期1246-1252,共7页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金面上项目(编号:82074493);河南省中医药学科领军人才项目(编号:豫卫中医函(2021)8号);河南省卫健委国家中医临床研究基地科研专项(编号:2022JDZX003);河南省中医药科学研究重大专项(编号:2023ZYZD02)。

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(RTPCR)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。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

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 

分 类 号：R96[医药卫生—药理学]