肾衰Ⅱ号方对慢性肾功能衰竭大鼠糖酵解限速酶HK2及PKM2表达的影响  

Effects of Shenshuai Formula Ⅱ on glycolysis rate-limiting enzymes HK2 and PKM2 expression in chronic renal failure rats

作　　者：杨柳易 王琛[1] 王蒙 周圆[1] YANG Liu-yi;WANG Chen;WANG Meng;ZHOU Yuan(Department of Nephrology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Nephropathy Institute of Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai Key Laboratory of Traditional Chinese Clinical Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院肾病科,上海中医药大学中医肾病研究所,上海市中医临床重点实验室,上海201203

出　　处：《中华中医药杂志》2020年第11期5807-5811,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81973770);上海市进一步加快中医药事业发展三年行动计划建设项目(No.Y2018-2020-FWTX-7005)。

摘　　要：目的:观察肾衰Ⅱ号方对5/6(A/I)慢性肾功能衰竭(CRF)大鼠肾组织糖酵解限速酶的影响及其作用机制。方法:随机选取55只SD雄性大鼠采用5/6(A/I)肾切除术制备CRF大鼠模型,将36只成功造模大鼠随机分为模型组、中药组、西药组,每组12只,另取12只大鼠作为假手术组。给予相应干预,8周后检测肌酐清除率(Ccr),采用比色法测定血清乳酸水平,RT-PCR检测糖酵解限速酶己糖激酶2(HK2)、丙酮酸激酶M2(PKM2)m RNA水平,Western blot检测HK2、PKM2和纤维连接蛋白(Fn)的表达,Masson染色观察肾组织病理形态。结果:与假手术组比较,模型组Ccr降低(P<0.01),Fn蛋白表达增加(P<0.01),HK2、PKM2 mRNA及蛋白表达增加(P<0.01,P<0.05);与模型组比较,中药组及西药组Fn蛋白表达降低(P<0.01),中药组HK2、PKM2 mRNA及蛋白表达降低(P<0.01,P<0.05)。Masson染色结果显示,与假手术组比较,模型组胶原沉积明显,肾间质纤维化严重(P<0.01);与模型组比较,中药组与西药组胶原沉积明显减少,肾间质纤维化改善(P<0.01)。结论:慢性肾功能衰竭与糖酵解之间具有正相关性。肾衰Ⅱ号方改善肾功能,抑制肾间质纤维化的机制可能与抑制糖酵解有关。Objective:To observe the effects and mechanism of Shenshuai FormulaⅡon the rate-limiting enzyme of renal glycolysis in rats with chronic renal failure induced by 5/6 ablation/infarction(A/I).Methods:Totally 55 SD male rats were randomly selected to induce CRF rat model by 5/6(A/I)and 36 successful rat models were randomized into model group,Chinese medicine group and Western medicine group,12 rats in each group.Another 12 rats were recruited as sham operation group.The corresponding intervention was given.After 8 weeks,Ccr was observed,the serum lactate level was determined by colorimetry.The mRNA levels of glycolysis rate-limiting enzymes hexokinase 2(HK2)and pyruvate kinase M2(PKM2)were detected by RTPCR,and HK2,PKM2 and Fn were detected by Western blot.The renal pathological changes were observed by Masson staining.Results:Compared with the sham operation group,the Ccr of the model group decreased(P<0.01).The expression of Fn protein in the model group increased(P<0.01),and the expression of HK2 and PKM2 mRNA and protein expression increased significantly(P<0.01,P<0.05).Compared with the model group,the levels of Fn protein in the Chinese medicine group and the western medicine group decreased(P<0.01),and the expression levels of HK2 and PKM2 mRNA and protein expression decreased(P<0.01,P<0.05).The results of Masson staining showed that compared with the sham operation group,the collagen deposition was obvious and the renal interstitial fibrosis was serious in the model group(P<0.01).Compared with the model group,the collagen deposition in the Chinese medicine group and western medicine group was significantly decreased,and the renal interstitial fibrosis was improved(P<0.01).Conclusion:There was a positive correlation between chronic renal failure and glycolysis.The mechanism of Shenshuai Formula II in improving renal function and inhibiting renal interstitial fibrosis may be related to the inhibition of glycolysis.

关 键 词：慢性肾功能衰竭 肾衰Ⅱ号方 糖酵解 己糖激酶2 丙酮酸激酶M2 

分 类 号：R285.5[医药卫生—中药学]