疏利三焦法通过转化生长因子-β途径调节糖尿病肾病足细胞凋亡机制的研究
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摘要
目的研究疏利三焦法通过转化生长因子-β(TGF-β)途径调节糖尿病肾病足细胞凋亡的机制。方法 48只SD大鼠,随机抽取8只作为对照组,余40只进行糖尿病肾病模型造模,取60 mg/kg的链脲佐菌素(STZ)溶解于0.1 mol/L灭菌枸橼酸钠缓冲液(pH 4.0)进行腹腔注射,对照组大鼠腹腔注射等量灭菌枸橼酸钠缓冲液。将34只造模成功大鼠随机分为模型组8只、缬沙坦组8只、疏利组9只、疏利高组9只。疏利组予三消汤9 mL/(kg·d)灌胃;疏利高组予三消汤18 mL/(kg·d)灌胃;缬沙坦组予缬沙坦10 mg/(kg·d)灌胃;对照组和模型组予0.9%氯化钠注射液18 mL/(kg·d)灌胃,连续给药8周。干预8周后,观察大鼠血糖、肌酐、尿素氮、24 h尿蛋白定量及肾脏TGF-β_1、Smad2/3蛋白、Smad7蛋白和足细胞凋亡水平。结果模型组、缬沙坦组、疏利组、疏利高组血糖、肌酐、尿素氮、24 h尿蛋白定量水平均高于对照组(P<0.05);缬沙坦组、疏利组、疏利高组肌酐、尿素氮、24 h尿蛋白定量,疏利组、疏利高组血糖与模型组比较均降低(P<0.05);疏利组、疏利高组血糖、肌酐、尿素氮、24 h尿蛋白定量均低于缬沙坦组(P<0.05);疏利高组血糖、肌酐、尿素氮、24 h尿蛋白定量均低于疏利组(P<0.05)。模型组、缬沙坦组、疏利组、疏利高组TGF-β_1、Smad2/3蛋白、足细胞凋亡均高于对照组(P<0.05),Smad7蛋白低于对照组(P<0.05);缬沙坦组、疏利组、疏利高组TGF-β_1、Smad2/3蛋白、足细胞凋亡均低于模型组(P<0.05),Smad7蛋白高于模型组(P<0.05);疏利组、疏利高组TGF-β_1、Smad2/3蛋白、足细胞凋亡均低于缬沙坦组(P<0.05),Smad7蛋白高于缬沙坦组(P<0.05);疏利高组TGF-β_1、Smad2/3蛋白、足细胞凋亡低于疏利组(P<0.05),Smad7蛋白高于疏利组(P<0.05)。结论疏利三焦法可通过抑制TGF-β/Smad信号通路激活,进而抑制足细胞凋亡,缓解糖尿病肾病病情,值得进一步研究探讨。
Objective To investigate the mechanism of Shuli Sanjiao method regulating podocyte apoptosis in diabetic nephropathy through transforming growth factor-β(TGF-β) pathway. Methods Of the 48 SD rats, 8 were randomly selected as control group, and the remaining 40 rats were used to establish diabetic nephropathy model, 60 mg/kg streptozotocin(STZ) was dissolved in 0.1 mol/L sterilized sodium citrate buffer(pH 4.0) for intraperitoneal injection. The rats in the control group were intraperitoneally injected with an equal amount of sterile sodium citrate buffer. 34 rats were successfully modeled and divided into model group(8 rats), valsartan group(8 rats), shuli group(9 rats) and shuligao group(9 rats). The shuli group was given Sanxiao decoction 9 mL/(kg·d) by gavage, the shuligao group was given Sanxiao decoction 18 mL/(kg·d) by gavage, the valsartan group was given valsartan 10 mg/(kg·d) by gavage, and the control group and the model group were given 0.9% sodium chloride injection 18 mL/(kg·d) by gavage. Continuous administration for 8 weeks, after 8 weeks of intervention, the levels of blood sugar, creatinine, urea nitrogen, 24-hour urinary protein, renal TGF-β_1, Smad2/3, Smad7 and podocyte apoptosis were observed. Results The levels of blood sugar, creatinine, urea nitrogen and 24-hour urinary protein in model group, valsartan group, shuli group and shuligao group were higher than those in control group(P<0.05). The levels of creatinine, urea nitrogen and 24-hour urinary protein in valsartan group, shuli group and shuligao group were lower than those in model group, the blood sugar in shuli group and shuligao group were lower than those in model group(P<0.05). The levels of blood sugar, creatinine, urea nitrogen and 24-hour urinary protein in shuli group and shuligao group were lower than those in valsartan group(P<0.05). The levels of blood sugar, creatinine, urea nitrogen and 24-hour urinary protein in shuligao group were lower than those in shuli group(P<0.05). The levels of TGF-β_1, Smad2/3 and podocyte apoptosis in model group were higher than those in control group(P<0.05), while the Smad7 was lower than that in control group(P<0.05). The levels of TGF-β_1, Smad2/3 and podocyte apoptosis in valsartan group, shuli group and shuligao group were lower than those in model group(P<0.05), while the Smad7 was higher than that in model group(P<0.05). The levels of TGF-β_1, Smad2/3 and podocyte apoptosis in shuli group and shuligao group were lower than those in valsartan group(P<0.05), while the Smad7 was higher than that invalsartan group(P<0.05). The levels of TGF-β_1, Smad2/3 and podocyte apoptosis in shuligao group were lower than those in shuli group(P<0.05), while the Smad7 was higher than that in shuli group(P<0.05). Conclusion The shuli sanjiao method can inhibit the activation of TGF-β/Smad signaling pathway, thereby inhibiting podocyte apoptosis and alleviating the condition of diabetic nephropathy, which deserves further study and discussion.
Objective To investigate the mechanism of Shuli Sanjiao method regulating podocyte apoptosis in diabetic nephropathy through transforming growth factor-β(TGF-β) pathway. Methods Of the 48 SD rats, 8 were randomly selected as control group, and the remaining 40 rats were used to establish diabetic nephropathy model, 60 mg/kg streptozotocin(STZ) was dissolved in 0.1 mol/L sterilized sodium citrate buffer(pH 4.0) for intraperitoneal injection. The rats in the control group were intraperitoneally injected with an equal amount of sterile sodium citrate buffer. 34 rats were successfully modeled and divided into model group(8 rats), valsartan group(8 rats), shuli group(9 rats) and shuligao group(9 rats). The shuli group was given Sanxiao decoction 9 mL/(kg·d) by gavage, the shuligao group was given Sanxiao decoction 18 mL/(kg·d) by gavage, the valsartan group was given valsartan 10 mg/(kg·d) by gavage, and the control group and the model group were given 0.9% sodium chloride injection 18 mL/(kg·d) by gavage. Continuous administration for 8 weeks, after 8 weeks of intervention, the levels of blood sugar, creatinine, urea nitrogen, 24-hour urinary protein, renal TGF-β_1, Smad2/3, Smad7 and podocyte apoptosis were observed. Results The levels of blood sugar, creatinine, urea nitrogen and 24-hour urinary protein in model group, valsartan group, shuli group and shuligao group were higher than those in control group(P<0.05). The levels of creatinine, urea nitrogen and 24-hour urinary protein in valsartan group, shuli group and shuligao group were lower than those in model group, the blood sugar in shuli group and shuligao group were lower than those in model group(P<0.05). The levels of blood sugar, creatinine, urea nitrogen and 24-hour urinary protein in shuli group and shuligao group were lower than those in valsartan group(P<0.05). The levels of blood sugar, creatinine, urea nitrogen and 24-hour urinary protein in shuligao group were lower than those in shuli group(P<0.05). The levels of TGF-β_1, Smad2/3 and podocyte apoptosis in model group were higher than those in control group(P<0.05), while the Smad7 was lower than that in control group(P<0.05). The levels of TGF-β_1, Smad2/3 and podocyte apoptosis in valsartan group, shuli group and shuligao group were lower than those in model group(P<0.05), while the Smad7 was higher than that in model group(P<0.05). The levels of TGF-β_1, Smad2/3 and podocyte apoptosis in shuli group and shuligao group were lower than those in valsartan group(P<0.05), while the Smad7 was higher than that invalsartan group(P<0.05). The levels of TGF-β_1, Smad2/3 and podocyte apoptosis in shuligao group were lower than those in shuli group(P<0.05), while the Smad7 was higher than that in shuli group(P<0.05). Conclusion The shuli sanjiao method can inhibit the activation of TGF-β/Smad signaling pathway, thereby inhibiting podocyte apoptosis and alleviating the condition of diabetic nephropathy, which deserves further study and discussion.
引文
[1] 陈关芬,辛丽芹.糖尿病肾病与合并糖尿病的非糖尿病肾病患者临床特征及预后比较[J].山东医药,2016,56(46):59-61.
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[5] 罗文娟,喻嵘,唐元,等.左归降糖益肾方对2型糖尿病肾病小鼠足细胞凋亡的影响[J].中医杂志,2015,56(6):511-514.
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[7] 尤丽菊,芦琨,田华.姜黄素对糖尿病肾病大鼠的肾转化生长因子表达影响[J].中国临床药理学杂志,2015,31(15):1530-1532.
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[10] 郑涛,李晓宁,王晓梅.宣通三焦汤治疗终末期糖尿病肾病临床研究[J].山东中医杂志,2014,33(6):449-450.
[11] 夏世澄.从三焦论治终末期糖尿病肾病34例[J].中国中医急症,2005,14(8):786.
[12] Shen Y,Cai R,Sun J,et al.Diabetes mellitus as a risk factor for incident chronic kidney disease and end-stage renal disease in women compared with men:a systematic review and meta-analysis[J].Endocrine,2017,55(1):66-76.
[13] Denhez B,Geraldes P.Regulation of nephrin phosphorylation in diabetes and chronic kidney injury[J].Adv Exp Med Biol,2017,966:149-161.
[14] 陈瑜,刘超.足细胞损伤与糖尿病肾病[J].中国实用内科杂志,2007,27(23):1876-1879.
[15] Ka SM,Yeh YC,Huang XR,et al.Kidney-targeting Smad7 gene transfer inhibits renal TGF-β/MAD homologue(SMAD)and nuclear factor κB(NF-κB)signalling pathways,and improves diabetic nephropathy in mice[J].Diabetologia,2012,55(2):509-519.
[16] 王宇晖,童孟立,杨汝春.黄芪甲苷联合大黄素通过Smads通路抑制TGF-β1诱导的足细胞凋亡[J].中华中医药学刊,2014,32(5):1089-1093.
[2] Chuang PY,Dai Y,Liu R,et al.Alteration of forkhead Box O(Foxo4)acetylation mediates apoptosis of podocytes in diabetes mellitus[J].Plos One,2011,6(8):e23566.
[3] 王静,郝丽,陈卫东.普罗布考对糖尿病肾病大鼠肾组织转化生长因子-β1及血红素加氧酶-1表达的影响[J].蚌埠医学院学报,2013,38(10):1229-1234.
[4] Eid AA.Mammalian target of rapamycin regulates Nox4-mediated podocyte depletion in diabetic renal injury[J].Diabetes,2013,62(8):2935-2947.
[5] 罗文娟,喻嵘,唐元,等.左归降糖益肾方对2型糖尿病肾病小鼠足细胞凋亡的影响[J].中医杂志,2015,56(6):511-514.
[6] 孙建新,郑蛟东,吕芳.试从三焦论治糖尿病肾病[J].北京中医药,2008,27(7):517-518.
[7] 尤丽菊,芦琨,田华.姜黄素对糖尿病肾病大鼠的肾转化生长因子表达影响[J].中国临床药理学杂志,2015,31(15):1530-1532.
[8] 徐叔云,卞如濂,陈修.药理实验方法学[M].3版.北京:人民卫生出版社,2002:1861.
[9] 金丽霞,金丽军,宋立群,等.基于三焦气化理论辨治糖尿病肾病[J].中医药信息,2017,34(1):44-46.
[10] 郑涛,李晓宁,王晓梅.宣通三焦汤治疗终末期糖尿病肾病临床研究[J].山东中医杂志,2014,33(6):449-450.
[11] 夏世澄.从三焦论治终末期糖尿病肾病34例[J].中国中医急症,2005,14(8):786.
[12] Shen Y,Cai R,Sun J,et al.Diabetes mellitus as a risk factor for incident chronic kidney disease and end-stage renal disease in women compared with men:a systematic review and meta-analysis[J].Endocrine,2017,55(1):66-76.
[13] Denhez B,Geraldes P.Regulation of nephrin phosphorylation in diabetes and chronic kidney injury[J].Adv Exp Med Biol,2017,966:149-161.
[14] 陈瑜,刘超.足细胞损伤与糖尿病肾病[J].中国实用内科杂志,2007,27(23):1876-1879.
[15] Ka SM,Yeh YC,Huang XR,et al.Kidney-targeting Smad7 gene transfer inhibits renal TGF-β/MAD homologue(SMAD)and nuclear factor κB(NF-κB)signalling pathways,and improves diabetic nephropathy in mice[J].Diabetologia,2012,55(2):509-519.
[16] 王宇晖,童孟立,杨汝春.黄芪甲苷联合大黄素通过Smads通路抑制TGF-β1诱导的足细胞凋亡[J].中华中医药学刊,2014,32(5):1089-1093.