基于析因设计探讨金雀根及肾消通络方对糖尿病肾病大鼠肾组织CD68/iNOS和p38MAPK信号通路的影响  

Discussion on the effects of root of caragana sinica and Shenxiao Tongluo Formula on CD68/iNOS and p38MAPK signaling pathway in nephridial tissue of diabetic kidney disease rats with factorial design

作　　者：聂浩坤 聂远 林健 丁英钧[1,2] NIE Hao-kun;NIE Yuan;LIN Jian;DING Ying-jun(Hebei University of Chinese Medicine,Shijiazhuang 050200,China;Hebei Key Laboratory of Integrative Medicine on Liver-Kidney Patterns,Shijiazhuang 050091,China)

机构地区：[1]河北中医学院,石家庄050200 [2]河北省中西医结合肝肾病证研究重点实验室,石家庄050091

出　　处：《中华中医药杂志》2023年第3期1235-1240,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：河北省自然科学基金项目(No.H2019423087)。

摘　　要：目的:基于析因设计观察金雀根及肾消通络方对糖尿病肾病大鼠肾组织巨噬细胞标记物CD68/诱导型一氧化氮合酶(iNOS)、p38MAPK信号通路的影响。方法:将70只SD大鼠随机分为假手术组(S组)10只和造模组60只。造模组大鼠以单侧肾切除联合一次性腹腔注射链脲霉素(STZ)建立糖尿病肾病模型。根据析因设计原理将成模大鼠随机分为模型组(A0B0组)、肾消通络方组(A1B1组)、金雀根组(A1B0组)、肾消通络方减金雀根组(A0B1组),另设阳性对照恩格列净组(Em组)。S组和A0B0组给予等容积的1%羧甲基纤维素钠溶液灌胃,其余组给予相应药物灌胃8周。治疗前及治疗后每隔4周检测空腹血糖、24 h尿微量白蛋白(U-mAlb)。药物干预8周后处死大鼠,检测血肌酐(Scr)、血尿素氮(BUN);PASM-Masson染色观察肾脏病理,免疫组化法检测肾组织CD68、iNOS表达;Western Blot检测肾组织中骨调素(OPN)、转化生长因子-β1(TGF-β1)、p38MAPK、p-p38MAPK/p38MAPK蛋白表达。结果:A0B0组大鼠空腹血糖、Scr、BUN和U-mAlb水平较S组显著升高(P<0.01),肾脏病理表现为肾小球基底膜增厚、系膜增生、系膜外基质增多、出现K-W结节,CD68、iNOS、OPN、TGF-β1、p-p38MAPK蛋白表达显著升高(P<0.01);经治疗后,Em组、A1B1组、A0B1组血清Scr、BUN、U-mAlb水平和肾组织CD68、iNOS、OPN、TGF-β1、p-p38MAPK/p38MAPK表达均有所下调,与A0B0组比较差异有统计学意义(P<0.05,P<0.01);Em组空腹血糖水平显著下调(P<0.01),A1B1组、A0B1组和A1B0组空腹血糖与A0B0组比较差异无统计学意义。结论:金雀根和肾消通络方均可改善肾功能,降低尿蛋白,减轻糖尿病肾病肾脏炎性损伤,其机制可能与抑制巨噬细胞和p38MAPK信号通路激活,减少TGF-β1等炎症介质的释放有关。Objective:To observe the effects of root of caragana sinica and Shenxiao Tongluo Formula on macrophage marker CD68/induced nitric oxide synthase(iNOS)and p38MAPK signaling pathway in nephridial tissue of diabetic kidney disease rats with factorial design.Methods:Seventy SD rats were randomly divided into sham-operated group(S group,n=10)and model group(n=60).The modeling group was established by unilateral nephrectomy combined with streptozotocin(STZ)as disposable intraperitoneal injection of diabetic kidney disease model.According to the principle of factorial design,the model rats were randomly divided into model group(A0B0),Shenxiao Tongluo Formula group(A1B1),root of caragana sinica group(A1B0),Shenxiao Tongluo Formula reducing root of caragana sinica group(A0B1),and positive control engram(Em)group.The sham-operated group and model group were given an equal volume of 1%carboxymethylcellulose sodium solution by gavage,and the remaining groups were given the corresponding drugs by gavage for 8 weeks.Fasting blood glucose(FBG)and 24 h urinary microalbumin(U-mAlb)were measured before and every 4 weeks after treatment.Rats were executed after 8 weeks of drug intervention,and blood creatinine(Scr)and blood urea nitrogen(BUN)were measured;PASM-Masson staining was performed to observe renal pathology,and CD68 and iNOS expression in renal tissues were detected by immunohistochemistry;OPN,TGF-β1,p38MAPK and p-p38MAPK protein expression in renal tissues were detected by Western Blot.Results:Serum FBG,Scr,BUN and U-mAlb levels were significantly upregulated in the A0B0 group of rats compared with the S group(P<0.01),and renal pathology showed glomerular basement membrane thickening,thylakoid hyperplasia,increased extrathylakoid matrix,the appearance of K-W nodules,and significantly higher expression of CD68,iNOS,OPN,TGF-β1,and p-p38MAPK/p38MAPK proteins(P<0.01);After treatment,serum Scr,BUN,U-mAlb levels and renal tissue CD68,iNOS,OPN,TGF-β1,p-p38MAPK/p38MAPK expression were downregulated in the Em,A1B1,A0B1 groups,w

关 键 词：金雀根 肾消通络方 糖尿病肾病 P38MAPK信号通路 炎症反应 析因设计 

分 类 号：R285.5[医药卫生—中药学]