丹酚酸B调节Bax/Bcl-2的表达对系膜增生性肾炎大鼠肾脏保护作用及机制
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摘要
目的 探究丹酚酸B(SAB)对大鼠系膜增生性肾炎(Ms PGN)肾脏保护作用及机制。方法 选择3周龄SD大鼠,采用改良慢性血清病法复制Ms PGN大鼠模型,分别设置健康对照组(Ctrl组)、SAB单独作用组(SAB组)、Ms PGN组和Ms PGN+SAB组,其中加药组采用口服100 mg/kg SAB。HE染色观察细胞形态,缺口末端标记(TUNEL)染色、免疫组织化学法和Western blot技术检测组织细胞凋亡及相关分子标记;ELISA检测肾损伤炎症因子;Western blot技术检测基因核因子κB p65(NF-κB p65)和肿瘤坏因子-α(TNF-α)等相关分子表达。结果 与Ctrl组比较,SAB单独作用于健康大鼠后,大鼠体质量、尿蛋白、血清肌酐及尿氮含量、细胞凋亡及相关分子、炎症因子无明显差异;与Ctrl组比较, Ms PGN组大鼠体质量无明显差异,尿蛋白、血清肌酐和尿氮含量,促细胞凋亡因子、炎症因子明显上升(P<0.05),凋亡抑制因子显著下降(P<0.05);与Ms PGN组比较,Ms PGN+SAB组大鼠体质量无明显变化,尿蛋白、血清肌酐和尿氮含量、促细胞凋亡因子、炎症因子显著下降(P<0.05),凋亡抑制因子显著上调(P<0.05)。结论 SAB在100 mg/kg剂量下对大鼠无明显毒性,并可以降低NF-κB p65和TNF-α表达,抑制肾小球细胞凋亡,降低大鼠肾小球炎症损伤。
Objective To investigate the function and mechanism of salvianolic acid B(SAB) on mesangial proliferative glomerulonephritis(Ms PGN) in rat models.Methods Three weeks SD rats were selected. The improved Ms PGN model rats were randomly divided into following groups: healthy control(Ctrl group); rats treated by the BSA only group(SAB group); rats treated by the BSA group(Ms PGN group); rats treated by the BSA and SAB group(Ms PGN+SAB group). The SAB was accepted 100 mg/kg by oral. HE staining was performed to detect the cell morphology; immunohistochemisty, TUNEL staining and Western blot were performed to investigate the apoptosis and markers; the level of inflammatory factors of renal injury were detected by ELISA; and Western blot also be used to detected the proteins of nuclear factor kappa-B p65(NF-κB p65) and tumor necrosis factor-α(TNF-α).Results Compared with the Ctrl group, the SAB group, there was no obvious difference in body weight, proteinuria, serum creatinine and urea nitrogen, and apoptosis-related markers and inflammatory. Compared with the Ctrl group, the Ms PGN group rats displayed no difference between body weight, but the proteinuria, serum creatinine and urea nitrogen, and apoptosis-related markers and pro-inflammatory were increased significantly(P<0.05).Compared with the Ms PGN group, the Ms PGN+SAB group rats had no difference between body weight, but the proteinuria, serum creatinine and urea nitrogen, and apoptosis-related markers and pro-inflammatory were decreased significantly(P<0.05).Conclusion SAB has no obvious toxic effect to rats in the dose of 100 mg/kg; and inhibits the expression of NF-κB p65 and TNF-α, decrease the apoptosis and inflammatory of glomerular cells in rat models.
Objective To investigate the function and mechanism of salvianolic acid B(SAB) on mesangial proliferative glomerulonephritis(Ms PGN) in rat models.Methods Three weeks SD rats were selected. The improved Ms PGN model rats were randomly divided into following groups: healthy control(Ctrl group); rats treated by the BSA only group(SAB group); rats treated by the BSA group(Ms PGN group); rats treated by the BSA and SAB group(Ms PGN+SAB group). The SAB was accepted 100 mg/kg by oral. HE staining was performed to detect the cell morphology; immunohistochemisty, TUNEL staining and Western blot were performed to investigate the apoptosis and markers; the level of inflammatory factors of renal injury were detected by ELISA; and Western blot also be used to detected the proteins of nuclear factor kappa-B p65(NF-κB p65) and tumor necrosis factor-α(TNF-α).Results Compared with the Ctrl group, the SAB group, there was no obvious difference in body weight, proteinuria, serum creatinine and urea nitrogen, and apoptosis-related markers and inflammatory. Compared with the Ctrl group, the Ms PGN group rats displayed no difference between body weight, but the proteinuria, serum creatinine and urea nitrogen, and apoptosis-related markers and pro-inflammatory were increased significantly(P<0.05).Compared with the Ms PGN group, the Ms PGN+SAB group rats had no difference between body weight, but the proteinuria, serum creatinine and urea nitrogen, and apoptosis-related markers and pro-inflammatory were decreased significantly(P<0.05).Conclusion SAB has no obvious toxic effect to rats in the dose of 100 mg/kg; and inhibits the expression of NF-κB p65 and TNF-α, decrease the apoptosis and inflammatory of glomerular cells in rat models.
引文
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[2] Wu Y Q,Wang Z F,Xu H F,et al.Frequency of primary glomerular disease in northeastern china[J].Braz J Med Biol Res,2011,44(8):810-3.
[3] Zhang A,Han Y F,Wang B,et al.Beyond gap junction channel function:The expression of cx43 contributes to aldosterone-induced mesangial cell proliferation via the erk1/2 and pkc pathways[J].Cell Physiol Biochem,2015,36(3):1210-22.
[4] Chen Y H,Du G H,Zhang J T.Salvianolic acid B protects brain against injuries caused by ischemia-reperfusion in rats[J].Acta Pharmacol Sin,2000,21(5):463-6.
[5] Tian J,Li G,Zhang S,et al.SMND-309,a novel derivate of salvianolic acid B,attenuates apoptosis and ameliorates mitochondrial energy metabolism in rat cortical neurons[J].Basic Clin Pharmacol Toxicol,2009,104(2):176-84.
[6] Striker L J,Doi T,Elliot S,et al.The contribution of glomerular mesangial cells to progressive glomerulosclerosis[J].Semin Nephrol,1989,9(4):318-28.
[7] Bajcsi D,Constantinou K,Krenacs L,et al.Rapidly progressive proliferative glomerulonephritis with monoclonal immunoglobulin G deposits despite the mild histological changes case report[J].Orv Hetil,2018,159(38):1567-72.
[8] Xu J,Wei K,Zhang G,et al.Ethnopharmacology,phytochemistry,and pharmacology of Chinese Salvia species:A review[J].J Ethnopharmacol,2018,225:18-30.
[9] Lin Y L,Wu C H,Luo M H,et al.In vitro protective effects of salvianolic acid B on primary hepatocytes and hepatic stellate cells[J].J Ethnopharmacol,2006,105(1-2):215-22.
[10] Tang M,Zhang J.Prostate apoptosis response-4 involved in the protective effect of salvianolic acid B against amyloid beta peptide-induced damage in PC12 cells[J].Jpn J Pharmacol,2002,88(4):422-7.
[11] Li Q,Verma I M.NF-kappaB regulation in the immune system[J].Nat Rev Immunol,2002,2(10):725-34.
[12] Vermeulen L,De Wilde G,Van Dame P,et al.Transcriptional activation of the NF-κB p65 subunit by mitogen-and stress-activated protein kinase-1 (MSK1)[J].EMBO J,2003,22(6):1313-24.
[13] Ishinaga H,Jono H,Lim J H,et al.Synergistic induction of nuclear factor-kappaB by transforming growth factor-beta and tumour necrosis factor-alpha is mediated by protein kinase A-dependent Rela Acetylation[J].Biochem J,2009,417(2):583-91.