益气健脾消瘀泄浊方对慢性肾衰竭大鼠骨骼肌自噬的影响

Effects of Yiqi Jianpi Xiaoyu Xiezhuo Decoction on Autophagy of Skeletal Muscle in Rats with Chronic Renal Failure

[目的]观察益气健脾消瘀泄浊方对慢性肾衰竭(chronic renal failure,CRF)大鼠骨骼肌自噬的影响,探讨益气健脾消瘀泄浊方在保护肾功能基础上改善CRF大鼠蛋白质-能量营养不良(protein-energy wasting,PEW)的作用和机制。[方法]选择25只无特定病原体(specified pathogen free,SPF)级6周龄雄性SD大鼠建立CRF-PEW模型,并分为模型对照组、阳性对照组、中药治疗组,另选择5只相同遗传背景的SD大鼠作为假手术组;阳性对照组予复方藁-酮酸片混悬液,中药治疗组予益气健脾消瘀泄浊方中药煎剂,模型对照组与假手术组均予等体积0.9%氯化钠注射液,连续灌胃8周。于治疗前后检测大鼠血肌酐(serum creatinine,Scr)、血尿素氮(blood urea nitrogen,BUN)、血清白蛋白(albumin,ALB)、24 h尿蛋白(24 hours urine protein,24 h Upr)水平;检测大鼠骨骼肌p62、微管相关蛋白轻链3A/3B (microtubule-associated protein light chain 3A/3B,LC3A/B)、Beclin-1以及叉头框转录因子O3a(forkhead-box transcription factor O3a,Foxo3a)基因及蛋白表达水平。[结果]与假手术组比较,模型对照组Scr、BUN和24 h Upr水平显著升高(P<0.01),ALB显著降低(P<0.05);骨骼肌p62基因及蛋白表达水平显著降低(P<0.01),LC3A/B、Beclin-1、Foxo3a基因及蛋白表达水平显著升高(P<0.01)。与模型对照组比较,中药治疗组Scr、BUN和24 h Upr水平降低(P<0.05,P<0.01),ALB升高(P<0.01);骨骼肌p62基因及蛋白表达水平升高(P<0.01),LC3A/B、Beclin-1、Foxo3a基因及蛋白表达水平降低(P<0.05,P<0.01)。[结论]益气健脾消瘀泄浊方可改善CRF大鼠的PEW状态,防止骨骼肌萎缩,其作用机制与抑制骨骼肌自噬有关。

[Objective]To observe the effects of Yiqi Jianpi Xiaoyu Xiezhuo Decoction on autophagy of skeletal muscle in rats with chronic renal failure(CRF),and investigate the effect and mechanism of Yiqi Jianpi Xiaoyu Xiezhuo Decoction on relieving protein-energy wasting(PEW)on the basis of protecting renal function in rats with CRF.[Methods]Twenty-five male specified pathogen free(SPF)SD rats aged 6 weeks were modeled in CRF-PEW and randomly divided into three groups including model control group,positive drug control group and Chinese medicine treatment group.Another 5 male SD rats with the same genetic background were used as sham operation group.Rats in positive drug control group were given compound藁-Ketonic acid tablet suspension,Chinese medicine treatment group was given Yiqi Jianpi Xiaoyu Xiezhuo Decoction,model control group and sham operation group were given equal volume 0.9%sodium chloride injection.Each group was kept treatment continually by gavage for 8 weeks.The blood and urine were collected for serum creatinine(Scr),blood urea nitrogen(BUN),albumin(ALB)and 24 hours urine protein(24 h Upr)before and after treatment.The skeletal muscle was collected for detection of gene and protein expression of p62,microtubule-associated protein light chain 3 A/3 B(LC3 A/B),Beclin-1 and forkhead-box transcription factor O3 a(Foxo3 a).[Results]Compared with sham operation group,Scr,BUN and 24 h Upr level in model control group were significantly increased(P<0.01),ALB levels was significantly decreased(P<0.05);in skeletal muscle,gene and protein expression levels of p62 were significantly decreased(P<0.01),LC3 A/B,Beclin-1 and Foxo3 a were significantly increased(P<0.01).Compared with model control group,Scr,BUN and 24 h Upr levels in Chinese medicine treatment group were decreased(P<0.05,P<0.01),ALB level was increased(P<0.01);in skeletal muscle,gene and protein expression levels of p62 were increased(P<0.01),LC3 A/B,Beclin-1 and Foxo3 a were decreased(P<0.05,P<0.01).[Conclusion]Yiqi Jianpi Xiaoyu Xiezhuo Decoction can improve PEW in rats with CRF and prevent muscular atrophy.The mechanism may be related to inhibiting the autophagy of skeletal muscle.