健脾清化方对单侧输尿管梗阻模型大鼠P38MAPK信号通路的影响
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摘要
目的:观察健脾清化方对单侧输尿管梗阻模型大鼠P38MAPK的影响,并探讨其作用机制。方法:SD雄性大鼠90只,随机分为假手术组,模型组,西药组和中药组,行UUO术制备单侧输尿管模型,并于术后7、14、21 d处死大鼠。生化法检测大鼠血肌酐、尿素氮; HE、Masson染色观察肾脏病理形态; WB测肾组织p38、p-p38、TGF-β1蛋白表达;免疫组化法观察肾组织p38蛋白阳性面积率。结果:与正常组比较,模型组SCr、BUN含量、p38、p-p38、TGF-β1表达、p38阳性面积率均升高(P <0. 01,P <0. 05);与模型组比较:氯沙坦钾组和健脾清化方组SCr、BUN含量、p38、p-p38、TGF-β1蛋白表达、p38阳性面积率均降低(P <0. 01,P <0. 05)。HE、Masson示模型组肾小管明显扩张,肾盂肾盏明显扩张。结论:健脾清化方能改善单侧输尿管梗阻模型大鼠的肾脏病理改变,对肾脏损害有一定的保护作用,其机制可能与其能下调SCr、BUN的含量、p38、p-p38、TGF-β1的蛋白表达有关。
Objective: To observe the effects of Jianpi Qinghua decoction on P38 MAPK in rats with unilateral ureteral obstruction and explore its mechanism. Methods: A total of 90 male SD rats were randomly divided into a sham operation group,a model group,a western medicine group and a traditional Chinese medicine group,and the unilateral ureteral model was made by UUO surgery,and the rats were sacrificed at 7,14 and 21 days after surgery. The serum creatinine and urea nitrogen were detected by biochemical method; the pathological morphology of kidney was observed by HE and Masson staining; the expression of p38,p-p38 and TGF-β1 in renal tissue was measured by WB; the positive area ratio of p38 protein in renal tissue was observed by immunohistochemistry. Results: Compared with the normal group,the content of SCr,BUN,the expression of p38,p-p38,TGF-β1 and the positive area ratio of p38 protein in the model group were all increased( P < 0. 01,P < 0. 05); Compared with the model group,the content of SCr,BUN,the protein expression of p38,p-p38,TGF-β1 and the positive area rate of p38 in both the western medicine group and the Jianpi Qinghua decoction group were all decreased( P < 0. 01,P < 0. 05). HE and Masson showed that the renal tubules and the renal pelvis in the model group were significantly expanded. Conclusion: Jianpi Qinghua decoction can improve the renal pathological changes in the model rats with unilateral ureteral obstruction,which has a certain protective effect on kidney damage,and the mechanism may be related to down-regulate the protein expression of SCr,BUN,p38,p-p38 and TGF-β1.
Objective: To observe the effects of Jianpi Qinghua decoction on P38 MAPK in rats with unilateral ureteral obstruction and explore its mechanism. Methods: A total of 90 male SD rats were randomly divided into a sham operation group,a model group,a western medicine group and a traditional Chinese medicine group,and the unilateral ureteral model was made by UUO surgery,and the rats were sacrificed at 7,14 and 21 days after surgery. The serum creatinine and urea nitrogen were detected by biochemical method; the pathological morphology of kidney was observed by HE and Masson staining; the expression of p38,p-p38 and TGF-β1 in renal tissue was measured by WB; the positive area ratio of p38 protein in renal tissue was observed by immunohistochemistry. Results: Compared with the normal group,the content of SCr,BUN,the expression of p38,p-p38,TGF-β1 and the positive area ratio of p38 protein in the model group were all increased( P < 0. 01,P < 0. 05); Compared with the model group,the content of SCr,BUN,the protein expression of p38,p-p38,TGF-β1 and the positive area rate of p38 in both the western medicine group and the Jianpi Qinghua decoction group were all decreased( P < 0. 01,P < 0. 05). HE and Masson showed that the renal tubules and the renal pelvis in the model group were significantly expanded. Conclusion: Jianpi Qinghua decoction can improve the renal pathological changes in the model rats with unilateral ureteral obstruction,which has a certain protective effect on kidney damage,and the mechanism may be related to down-regulate the protein expression of SCr,BUN,p38,p-p38 and TGF-β1.
引文
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[14]Liu HS,Pan CE,Liu QG,et al.Effect of NF-kappaB and p38 MAPKin activated monocytes/macrophages on pro-inflammatory cytokines of rats with acute pancreatitis[J].World J Gastroenterol,2003,9(11):2513-2518.
[15]宋小乐.TGF-β1/Smad信号转导通路对肾间质纤维化的影响[J].南昌大学学报:医学版,2011,51(6):92-95,98.
[16]娄强,张芳,李淑莲.p38 MAPK在顺铂诱导肾小管上皮细胞凋亡中的作用[J].中国病理生理杂志,2018,34(9):1678-1683.
[2]Klahr S,Morrissey J.Obstructive nephropathy and renal fibrosis[J].Am J Physiol Renal Physiol,2002,283(5):F861-F875.
[3]Svensson CI,Marsala M,Westerlund A,et al.Activation of p38 mitogen-activated protein kinase in spinal microglia is a critical link in inflammation-induced spinal pain processing[J].J Neurochem,2003,86(6):1534-1544.
[4]杜俊英,方剑乔,梁宜,等.p38MAPK通路参与炎性痛及电针干预的可能性[J].医学信息(上旬刊),2011,24(6):3612-3613.
[5]杨晓溪,崔建美,喇孝瑾,等.针刺对哮喘大鼠肺组织p38蛋白激酶及c-fos蛋白表达的影响[J].时珍国医国药,2013,24(4):1004-1006.
[6]陈刚,何立群.健脾清化方治疗慢性肾衰竭53例临床观察[J].中国中西医结合肾病杂志,2006,7(10):591-593.
[7]Ahmed H,Mohamed Y,Tawfek A,et al.Renal fibrosis[J].Menoufia Med J,2015,28(2):540-546.
[8]袁媛.大鼠单侧输尿管结扎模型和饥饿模型尿液蛋白质组研究[D].北京:北京协和医学院,2015.
[9]张盟.CHOP基因敲除后抑制UUO诱导的小鼠肾脏纤维化的机制研究[D].武汉:华中科技大学,2016.
[10]邹朝霞,李均.内质网应激与肾脏纤维化[J].海南医学,2017,28(4):615-617.
[11]马晓红,王东,王云满,等.健脾清化方对慢性肾衰大鼠肾组织TGF-β1 mRNA及CTGFmRNA表达的影响[J].中国中医药科技,2013,20(2):122-123,128.
[12]徐蝶衣,王骞,张长明.健脾清化法治疗慢性肾衰的研究进展[J].环球中医药,2018,11(2):312-315.
[13]Zheng SY,Fu XB,Xu JG,et al.Inhibition of p38 mitogen-activated protein kinase may decrease intestinal epithelial cell apoptosis and improve intestinal epithelial barrier function after ischemia-reperfusion injury[J].World J Gastroenterol,2005,11(5):656-660.
[14]Liu HS,Pan CE,Liu QG,et al.Effect of NF-kappaB and p38 MAPKin activated monocytes/macrophages on pro-inflammatory cytokines of rats with acute pancreatitis[J].World J Gastroenterol,2003,9(11):2513-2518.
[15]宋小乐.TGF-β1/Smad信号转导通路对肾间质纤维化的影响[J].南昌大学学报:医学版,2011,51(6):92-95,98.
[16]娄强,张芳,李淑莲.p38 MAPK在顺铂诱导肾小管上皮细胞凋亡中的作用[J].中国病理生理杂志,2018,34(9):1678-1683.