[1]徐瑞,张晓,刘文霞.黄芪颗粒联合微RNA-30a对肾病综合征小鼠肾组织中内质网应激相关蛋白、有丝分裂原活化蛋白激酶蛋白和血清炎症因子水平的影响[J].新乡医学院学报,2023,40(11):1008-1015.[doi:10.7683/xxyxyxb.2023.11.002]
　XU Rui,ZHANG Xiao,LIU Wenxia.Effect of Huangqi granules combined with microRNA-30a on endoplasmic reticulum stress related protein,mitogen-activated protein kinase protein in renal tissue and serum inflammatory factor levels of mice with nephrotic syndrome[J].Journal of Xinxiang Medical University,2023,40(11):1008-1015.[doi:10.7683/xxyxyxb.2023.11.002]
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黄芪颗粒联合微RNA-30a对肾病综合征小鼠肾组织中内质网应激相关蛋白、有丝分裂原活化蛋白激酶蛋白和血清炎症因子水平的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
40卷
期数:
2023年11
页码:
1008-1015
栏目:
基础研究
出版日期:
2023-11-05

文章信息/Info

Title:
Effect of Huangqi granules combined with microRNA-30a on endoplasmic reticulum stress related protein,mitogen-activated protein kinase protein in renal tissue and serum inflammatory factor levels of mice with nephrotic syndrome
作者:
徐瑞; 张晓; 刘文霞
（南阳市第一人民医院儿三科，河南　南阳　473000）
Author(s):
XU Rui; ZHANG Xiao; LIU Wenxia
（Department of Pediatrics Ⅲ,Nanyang First People′s Hospital,Nanyang 473000,Henan Province,China）
关键词:
黄芪颗粒; 微RNA-30a; 肾病综合征; 小鼠; 内质网应激; 炎症反应; 有丝分裂原活化蛋白激酶蛋白
Keywords:
Huangqi granules; microRNA-30a; nephrotic syndrome; mice; endoplasmic reticulum stress; inflammatory response; mitogen-activated protein kinase protein
分类号:
R692
DOI:
10.7683/xxyxyxb.2023.11.002
文献标志码:
A
摘要:
目的　探讨黄芪颗粒联合微RNA（miR）-30a对肾病综合征小鼠内质网应激、炎症水平和有丝分裂原活化蛋白激酶（MAPK）蛋白表达的影响。
方法　将50只小鼠随机分为对照组（n=10）和模型组(n=40)。模型组小鼠经尾静脉注射阿霉素溶液（7.5 mg·kg-1）制备原发性肾病综合征（PNS）模型，对照组小鼠经尾静脉注射等体积的生理盐水。造模成功后将模型组小鼠随机分为PNS组、黄芪组、miR-30a组和黄芪+miR-30a组，每组10只。黄芪组小鼠灌胃黄芪颗粒冲剂（20 mg·g-1），每日2次；miR-30a组小鼠经尾静脉注射miR-30a antagomir（10 mg·kg-1），每周1次；黄芪+miR-30a组小鼠灌胃黄芪颗粒冲剂（20 mg·g-1，每周1次)，同时经尾静脉注射miR-30a antagomir（10 mg·kg-1）；模型组和对照组小鼠灌胃与黄芪组等量的生理盐水，每日1次；各组小鼠均干预4周。于给药0、2、4周时检测各组小鼠24 h尿蛋白；于末次给药24 h后，应用全自动生物化学分析仪检测各组小鼠血清中总胆固醇（TC）、三酰甘油（TG）、血尿素氮（BUN）和血肌酐（Scr）水平，采用酶联免疫吸附试验法检测各组小鼠血清中白细胞介素（IL）-6、IL-8和肿瘤坏死因子-α（TNF-α）水平，苏木精-伊红染色观察各组小鼠肾组织病理学变化，免疫组织化学法检测各组小鼠肾组织中糖调节蛋白-78（GRP-78）和激活转录因子6（ATF6）的表达，Western blot法检测各组小鼠肾组织中GRP-78、ATF6、p38丝裂原活化蛋白激酶（p38 MAPK）、磷酸化p38丝裂原活化蛋白激酶（p-p38 MAPK）、线粒体外活化蛋白激酶（ERK）、磷酸化线粒体外活化蛋白激酶（p-ERK）蛋白表达。
结果　PNS组、黄芪组、miR-30a组和黄芪+miR-30a组小鼠给药0、2、4周时24 h尿蛋白显著高于对照组（P<0.05）。给药2、4周时，黄芪组、miR-30a组和黄芪+miR-30a组小鼠的24 h尿蛋白显著低于PNS组（P<0.05）；黄芪组与miR-30a组小鼠的24 h尿蛋白比较差异无统计学意义（P>0.05）；黄芪+miR-30a组小鼠24 h尿蛋白显著低于黄芪组和miR-30a组（P<0.05）。PNS组、黄芪组、miR-30a组和黄芪+miR-30a组小鼠血清中TC、TG、BUN、Scr水平显著高于对照组（P<0.05）；黄芪组、miR-30a组和黄芪+miR-30a组小鼠血清中TC、TG、BUN、Scr水平显著低于PNS组（P<0.05）；黄芪组与miR-30a组小鼠血清中TC、TG、BUN、Scr水平比较差异无统计学意义（P>0.05）；黄芪+miR-30a组小鼠血清中TC、TG、BUN、Scr水平显著低于黄芪组和miR-30a组（P<0.05）。PNS组、黄芪组、miR-30a组和黄芪+miR-30a组小鼠血清中IL-6、IL-8、TNF-α水平显著高于对照组（P<0.05）；黄芪组、miR-30a组和黄芪+miR-30a组小鼠血清中IL-6、IL-8、TNF-α水平显著低于PNS组（P<0.05）；黄芪组与miR-30a组小鼠血清中IL-6、IL-8、TNF-α水平比较差异无统计学意义（P>0.05）；黄芪+miR-30a组小鼠血清中IL-6、IL-8、TNF-α水平显著低于黄芪组和miR-30a组（P<0.05）。对照组小鼠肾小球排列规则，无明显炎症细胞浸润；PNS组小鼠肾小球排列不规则，有大量炎症细胞浸润到肾间质；与PNS组相比，黄芪组和miR-30a组小鼠肾组织病理学变化得到明显改善，肾间质炎症细胞浸润明显减少；黄芪+miR-30a组小鼠肾间质偶见炎症细胞。PNS组、黄芪组、miR-30a组和黄芪+miR-30a组小鼠肾组织中GRP-78、ATF6蛋白相对表达量及p-p38 MAPK/p38 MAPK、p-ERK/ERK比值显著高于对照组（P<0.05）；黄芪组、miR-30a组和黄芪+miR-30a组小鼠肾组织中GRP-78、ATF6蛋白相对表达量及p-p38 MAPK/p38 MAPK、p-ERK/ERK比值显著低于PNS组（P<0.05）；黄芪组与miR-30a组小鼠肾组织中GRP-78、ATF6蛋白相对表达量及p-p38 MAPK/p38 MAPK、p-ERK/ERK比值比较差异无统计学意义（P>0.05）；黄芪+miR-30a组小鼠肾组织中GRP-78、ATF6蛋白相对表达量及p-p38 MAPK/p38 MAPK、p-ERK/ERK比值显著低于黄芪组和miR-30a组（P<0.05）。
结论　黄芪颗粒联合miR-30a可显著降低肾病综合征小鼠内质网应激，减轻炎症反应，抑制肾组织中MAPK蛋白的表达。
Abstract:
Objective　Effect of Huangqi granules combined with microRNA (miR)-30a on endoplasmic reticulum stress,inflammatory levels and mitogen-activated protein kinase (MAPK) protein expression in mice with nephrotic syndrome.
Methods　Fifty mice were randomly divided into control group (n=10) and model group (n=40).The mice in the model group were intravenously injected with adriamycin solution (7.5 mg·kg-1) to establish primary nephrotic syndrome (PNS) model,the mice in the control group were intravenously injected with equivalent volume of physiological saline.After successful modeling,the mice in the model group were randomly divided into the PNS group,Huangqi group,miR-30a group and Huangqi+miR-30a group,with 10 mice in each group.The mice in Huangqi group were given Huangqi particles (20 mg·g-1) by gavage twice a day;the mice in miR-30a group were intravenously injected with miR-30a antagomir (10 mg·kg-1) once a week;the mice in Huangqi+miR-30a group were given Huangqi particles (20 mg·g-1) by gavage once a week and intravenously injected with miR-30a antagomir (10 mg·kg-1) at the same time;the mice in the model group and control group were given equivalent volume of physiological saline by gavage once a day;all mice were treated for 4 weeks.At 0,2 and 4 weeks of administration,the levels of 24-hour urinary protein of mice in each group were detected.At 24 hours after the last administration the serum total cholesterol (TC),triglycerides (TG),blood urea nitrogen (BUN),serum creatinine (Scr) levels of mice in each group were measured by automatic biochemical analyzer;the levels of interleukin (IL)-6,IL-8 and tumor necrosis factor-α(TNF-α) in serum of mice in each group were detected by enzyme-linked immunosorbent assay;the renal pathological changes of mice in each group were observed by hematoxylin-eosin staining;the expressions of glucose-regulated protein-78 (GRP-78) and activating transcription factor 6 (ATF6) in renal tissues of the mice in each group were detected by immunohistochemistry;the expressions of GRP-78,ATF6,p38 mitogen-activated protein kinase (p38 MAPK),phosphorylated p38 mitogen-activated protein kinase(p-p38 MAPK),extracellular signal-regulated kinase (ERK) and phosphorylated extracellular signal-regulated kinase (p-ERK) in renal tissues of the mice in each group were detected by Western blot.
Results　At 0,2,4 weeks of administration,the level of 24-hour urinary protein of mice in the PNS group,Huangqi group,miR-30a group,Huangqi+miR-30a group was significantly higher than that in the control group (P<0.05);at 2,4 weeks of administration,the level of 24-hour urinary protein of mice in the Huangqi group,miR-30a group,Huangqi+miR-30a group was significantly lower than that in the PNS group (P<0.05);there was no significant difference in the level of 24-hour urinary protein of mice between the Huangqi group and miR-30a group(P>0.05);the level of 24-hour urinary protein of mice in the Huangqi+miR-30a group was significantly lower than that in the Huangqi group and miR-30a group (P<0.05).The levels of serum TC,TG,BUN and Scr of mice in the PNS group,Huangqi group,miR-30a group and Huangqi+miR-30a group were significantly higher than those in the control group (P<0.05);the levels of serum TC,TG,BUN and Scr of mice in the Huangqi group,miR-30a group and Huangqi+miR-30a group were significantly lower than those in the PNS group (P<0.05);there was no significant difference in the serum TC,TG,BUN and Scr levels of mice between the Huangqi group and the miR-30a group (P>0.05);the levels of serum TC,TG,BUN and Scr of mice in the Huangqi+miR-30a group were significantly lower than those in the Huangqi group and miR-30a group (P<0.05).The levels of serum IL-6,IL-8,TNF-α of mice in the PNS group,Huangqi group,miR-30a group and Huangqi+miR-30a group were significantly higher than those in the control group (P<0.05);the levels of serum IL-6,IL-8,TNF-α of mice in the Huangqi group,miR-30a group and Huangqi+miR-30a group were significantly lower than those in the PNS group (P<0.05);there was no significant difference in the serum IL-6,IL-8,TNF-α levels of mice between the Huangqi group and the miR-30a group (P>0.05);the levels of serum IL-6,IL-8,TNF-α of mice in the Huangqi+miR-30a group were significantly lower than those in the Huangqi group and miR-30a group (P<0.05).In the control group,the arrangement of glomerulus of mice were regular and without significant infiltration of inflammatory cells;in the PNS group,the arrangement of glomerulus of mice were irregular and with a large number of inflammatory cells infiltrated into the renal interstitium;compared with the PNS group,the pathological changes of renal tissue of the mice in the Huangqi group and miR-30a group significantly improved,and the infiltration of inflammatory cells in the renal interstitium significantly reduced;in the Huangqi+miR-30a group,inflammatory cells were occasionally seen in the renal interstitium of mice.The relative expressions of GRP-78,ATF6 protein and the ratios of p-p38 MAPK/p38 MAPK,p-ERK/ERK in renal tissue of the mice in the PNS group,Huangqi group,miR-30a and Huangqi+miR-30a group were significantly higher than those in the control group (P<0.05);the relative expressions of GRP-78,ATF6 protein and the ratios of p-p38 MAPK/p38 MAPK,p-ERK/ERK in the renal tissue of mice in the Huangqi group,miR-30a group and Huangqi+miR-30a group were significantly lower than those in the PNS group(P<0.05);there was no significant difference in the relative expressions of GRP-78,ATF6 protein and the ratios of p-p38 MAPK/p38 MAPK,p-ERK/ERK in renal tissue of the mice between the Huangqi group and the miR-30a group (P>0.05);the relative expression of GRP-78,ATF6 protein and the ratios of p-p38 MAPK/p38 MAPK,p-ERK/ERK in renal tissue of the mice in the Huangqi+miR-30a group were significantly lower than those in the Huangqi group and the miR-30a group (P<0.05).
Conclusion　Huangqi granules combined with miR-30a can significantly reduce endoplasmic reticulum stress and alleviate inflammatory reactions in mice with nephrotic syndrome.It also can significantly inhibit the expression of MAPK protein in renal tissue.

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更新日期/Last Update: 2023-11-05