肾康注射液对CAPD小鼠腹膜间皮细胞的保护作用及机制研究

目的探讨肾康注射液对连续性不卧床腹膜透析(CAPD)小鼠腹膜间皮细胞(PMCs)的保护作用及可能机制。方法40只ICR小鼠均分为空白对照(A)组,阳性对照腹透(B)组(2.5%腹透液),低(C)、中(D)、高(E)剂量肾康干预组(2.5%腹透液+5、10及20mL/kg肾康注射液),观察至4周。采用生化法检测空腹血糖、总胆固醇、三酰甘油、C-反应蛋白(CRP)水平;采用酶联免疫吸附试验检测血清及透出液中肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)、血管内皮生长因子(VEGF)、结缔组织生长因子(CTGF)水平;HE染色观察腹膜组织病理改变;分别以免疫组化染色和Real-t...

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Published in天津医药 Vol. 45; no. 8; pp. 860 - 864
Main Author 王荔;夏天;张洪震;李荣
Format Journal Article
LanguageChinese
Published 天津医科大学第二医院肾内科 300211%天津医科大学第二医院检验科 300211 2017
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ISSN0253-9896
DOI10.11958/20170502

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Abstract 目的探讨肾康注射液对连续性不卧床腹膜透析(CAPD)小鼠腹膜间皮细胞(PMCs)的保护作用及可能机制。方法40只ICR小鼠均分为空白对照(A)组,阳性对照腹透(B)组(2.5%腹透液),低(C)、中(D)、高(E)剂量肾康干预组(2.5%腹透液+5、10及20mL/kg肾康注射液),观察至4周。采用生化法检测空腹血糖、总胆固醇、三酰甘油、C-反应蛋白(CRP)水平;采用酶联免疫吸附试验检测血清及透出液中肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)、血管内皮生长因子(VEGF)、结缔组织生长因子(CTGF)水平;HE染色观察腹膜组织病理改变;分别以免疫组化染色和Real-timePCR检测腹膜组织中TNF-α、TGF-β1、VEGF、CTGF的蛋白质表达和mRNA转录水平。结果各组小鼠体质量、空腹血糖、总胆固醇、三酰甘油水平差异无统计学意义。与B组比较,C、E组差异均无统计学意义,但D组CRP水平显著减低,无论是血清还是腹腔透出液中,C、D组TNF-α、TGF-β1、VEGF及CTGF表达水平均降低,而E组TNF-α、TGF-β1升高(P<0.05),但E组VEGF及CTGF差异均无统计学意义。与E组比较,除C组腹腔液中CTGF差异无统计学意义外,血清及腹腔透出液中C、D组TNF-α、TGF-β1、VEGF、CTGF均降低(P<0.05)。B组可见腹膜间皮细胞均有损害;C、D、E组有不同程度改善。与B组比较,C、D、E组TNF-α、TGF-β1、VEGF、CTGF蛋白及mRNA水平呈逐渐降低趋势(P<0.05)。结论一定浓度的肾康注射液可通过抑制CAPD小鼠PMCs的TNF-α、TGF-β1、VEGF、CTGF的表达,保护PMCs,从而控制腹膜纤维化的发生发展。
AbstractList R459.51%R2-031; 目的 探讨肾康注射液对连续性不卧床腹膜透析(CAPD)小鼠腹膜间皮细胞(PMCs)的保护作用及可能机制.方法 40只ICR小鼠均分为空白对照(A)组,阳性对照腹透(B)组(2.5%腹透液),低(C)、中(D)、高(E)剂量肾康干预组(2.5%腹透液+5、10及20 mL/kg肾康注射液),观察至4周.采用生化法检测空腹血糖、总胆固醇、三酰甘油、C-反应蛋白(CRP)水平;采用酶联免疫吸附试验检测血清及透出液中肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)、血管内皮生长因子(VEGF)、结缔组织生长因子(CTGF)水平;HE染色观察腹膜组织病理改变;分别以免疫组化染色和Real-time PCR检测腹膜组织中TNF-α、TGF-β1、VEGF、CTGF的蛋白质表达和mRNA转录水平.结果 各组小鼠体质量、空腹血糖、总胆固醇、三酰甘油水平差异无统计学意义.与B组比较,C、E组差异均无统计学意义,但D组CRP水平显著减低,无论是血清还是腹腔透出液中,C、D组TNF-α、TGF-β1、VEGF及CTGF表达水平均降低,而E组TNF-α、TGF-β1升高(P<0.05),但E组VEGF及CTGF差异均无统计学意义.与E组比较,除C组腹腔液中CTGF差异无统计学意义外,血清及腹腔透出液中C、D组TNF-α、TGF-β1、VEGF、CTGF均降低(P<0.05).B组可见腹膜间皮细胞均有损害;C、D、E组有不同程度改善.与B组比较,C、D、E组TNF-α、TGF-β1、VEGF、CTGF蛋白及mRNA水平呈逐渐降低趋势(P<0.05).结论 一定浓度的肾康注射液可通过抑制CAPD小鼠PMCs的TNF-α、TGF-β1、VEGF、CTGF的表达,保护PMCs,从而控制腹膜纤维化的发生发展.
目的探讨肾康注射液对连续性不卧床腹膜透析(CAPD)小鼠腹膜间皮细胞(PMCs)的保护作用及可能机制。方法40只ICR小鼠均分为空白对照(A)组,阳性对照腹透(B)组(2.5%腹透液),低(C)、中(D)、高(E)剂量肾康干预组(2.5%腹透液+5、10及20mL/kg肾康注射液),观察至4周。采用生化法检测空腹血糖、总胆固醇、三酰甘油、C-反应蛋白(CRP)水平;采用酶联免疫吸附试验检测血清及透出液中肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)、血管内皮生长因子(VEGF)、结缔组织生长因子(CTGF)水平;HE染色观察腹膜组织病理改变;分别以免疫组化染色和Real-timePCR检测腹膜组织中TNF-α、TGF-β1、VEGF、CTGF的蛋白质表达和mRNA转录水平。结果各组小鼠体质量、空腹血糖、总胆固醇、三酰甘油水平差异无统计学意义。与B组比较,C、E组差异均无统计学意义,但D组CRP水平显著减低,无论是血清还是腹腔透出液中,C、D组TNF-α、TGF-β1、VEGF及CTGF表达水平均降低,而E组TNF-α、TGF-β1升高(P&lt;0.05),但E组VEGF及CTGF差异均无统计学意义。与E组比较,除C组腹腔液中CTGF差异无统计学意义外,血清及腹腔透出液中C、D组TNF-α、TGF-β1、VEGF、CTGF均降低(P&lt;0.05)。B组可见腹膜间皮细胞均有损害;C、D、E组有不同程度改善。与B组比较,C、D、E组TNF-α、TGF-β1、VEGF、CTGF蛋白及mRNA水平呈逐渐降低趋势(P<0.05)。结论一定浓度的肾康注射液可通过抑制CAPD小鼠PMCs的TNF-α、TGF-β1、VEGF、CTGF的表达,保护PMCs,从而控制腹膜纤维化的发生发展。
Abstract_FL Objective To study the protective effect of Shenkang injection on peritoneal mesothelial cells (PMCs) of continuous ambulatory peritoneal dialysis (CAPD) mice, and explore the possible mechanism. Methods Forty ICR mice were randomly divided into blank control (A) group, peritoneal dialysis (B) group, low dose of Shenkang (C) group ( 2.5%dialysate+5 mL/kg Shenkang injection), medium (D) group (2.5% dialysate+10 mL/kg Shenkang injection) and high (E) group (2.5% dialysate+ 20 mL/kg Shenkang injection). Mice were observed for 4 weeks. Fasting blood glucose, total cholesterol, triglyceride and C-reactive protein (CRP) were detected by biochemical assay. Tumor necrosis factor-alpha (TNF-α), transforming growth factor-beta 1 (TGF-β1), vascular endothelial growth factor (VEGF) and connective tissue growth factor (CTGF) levels of serum and dialysate were detected by ELISA. Pathological changes of peritoneal tissue were observed by HE staining. Expression and mRNA transcription levels of these four cytokines in the peritoneal tissue were detected by immunohistochemical staining and real-time PCR respectively. Results There were no significant differences in body weight, fasting blood glucose, total cholesterol and triglyceride between 5 groups of mice (P>0.05). Compared with group B, there was no significant difference in CRP level between group C and group E, but which was significantly decreased in group D (P<0.05). The serum and dialysate levels of TNF-α, TGF-β1, VEGF and CTGF were decreased in group C and group D. The serum and dialysate levels of TNF-αand TGF-β1 were significantly increased in group E (P<0.05), but there was no significant difference between VEGF and CTGF in group E. Compared with group E, except for CTGF in dialysate of group C, the serum and dialysate levels of TNF-α, TGF-β1, VEGF and CTGF were significant decreased in group C and group D (P<0.05). Damaged PMCs were found in group B, which were improved in various degrees in group C, group D and group E. Compared with group B, the protein expression and mRNA relative transcription levels of TNF-α, TGF-β1, VEGF and CTGF tended to decrease gradually in group C, group D and group E (P<0.05). Conclusion A certain concentration of Shenkang injection can protect PMCs by inhibiting the expression of TNF-α, TGF-β1, VEGF and CTGF in CAPD mice, so as to control the occurrence and development of peritoneal fibrosis.
Author 王荔;夏天;张洪震;李荣
AuthorAffiliation 天津医科大学第二医院肾内科,300211;检验科
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LI Rong
XIA Tian
WANG Li
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DocumentTitleAlternate Protective effect and its mechanism of Shenkang injection on peritoneal mesothelial cells in CAPD mice
DocumentTitle_FL Protective effect and its mechanism of Shenkang injection on peritoneal mesothelial cells in CAPD mice
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Keywords 肾功能衰竭
腹膜透析,持续不卧床
retroperitoneal fibrosis
peritoneal mesothelial cells
kidney failure
Shenkang injection
腹膜后纤维化
disease models
肾康注射液
continuous ambulatory
animal
peritoneal dialysis
腹膜间皮细胞
疾病模型,动物
Language Chinese
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Notes peritoneal dialysis, continuous ambulatory;kidney failure;retroperitoneal fibrosis;disease models, animal;peritoneal mesothelial cells; Shenkang injection
12-1116/R
Objective To study the protective effect of Shenkang injection on peritoneal mesothelial cells(PMCs)ofcontinuous ambulatory peritoneal dialysis(CAPD)mice,and explore the possible mechanism.Methods Forty ICR micewere randomly divided into blank control(A)group,peritoneal dialysis(B)group,low dose of Shenkang(C)group(2.5%dialysate+5mL/kg Shenkang injection),medium(D)group(2.5%dialysate+10mL/kg Shenkang injection)and high(E)group(2.5%dialysate+20mL/kg Shenkang injection).Mice were observed for4weeks.Fasting blood glucose,totalcholesterol,triglyceride and C-reactive protein(CRP)were detected by biochemical assay.Tumor necrosis factor-alpha(TNF-α),transforming growth factor-beta1(TGF-β1),vascular endothelial growth factor(VEGF)and connective tissuegrowth factor(CTGF)levels of serum and dialysate were detected by ELISA.Pathological changes of peritoneal t
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R459.51%R2-031; 目的 探讨肾康注射液对连续性不卧床腹膜透析(CAPD)小鼠腹膜间皮细胞(PMCs)的保护作用及可能机制.方法 40只ICR小鼠均分为空白对照(A)组,阳性对照腹透(B)组(2.5%腹透液),低(C)、中(D)、高(E)剂量肾康干预组(2.5%腹透液+5、10及20...
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SubjectTerms 腹膜透析,持续不卧床;肾功能衰竭;腹膜后纤维化;疾病模型,动物;腹膜间皮细胞;肾康注射液
Title 肾康注射液对CAPD小鼠腹膜间皮细胞的保护作用及机制研究
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