OBJECTIVE:To investigate the efficacy of Bushen Kangshuai(BS-KS)tablet on autophagy and polarization in mouse macrophage RAW 264.7.MEYHODS:Macrophage autophagy was induced by oxidized low-density lipoprotein(100μg/m ...OBJECTIVE:To investigate the efficacy of Bushen Kangshuai(BS-KS)tablet on autophagy and polarization in mouse macrophage RAW 264.7.MEYHODS:Macrophage autophagy was induced by oxidized low-density lipoprotein(100μg/m L).To detect the levels of autophagy,macrophage were transfected with double fluorescence LC3 autophagy adenovirus,then the numbers of autophagosomes and autophagic lysosomes were asessed by confocal microscopy.The autophagy related proteins expression of PI3 K,Akt,phospho-m Akt(p-Akt)and m TOR,phospho-m TOR(p-TOR),p62,microtubule-associated protein 1(LC3-Ⅱ)were determined by western blotting.The macrophage polarization model was induced by lipopolysaccharide(1μg/m L).The m RNA levels of i NOS,CD86(M1 macrophages marker molecules),and CD206,Arg-1(M2 macrophages marker molecules)were detected by real-time quantitative PCR.The concentration of cytokines TNF-αand IL-10 was determined by enzyme-linked immunosorbent assay.The protein expression of nuclear proteins PPAR-γ,NF-κB,and cytoplasmic protein IKBαwas determined by western blotting.RESULTS:The expression of the autophagy-related protein LC3-Ⅱwas increased and the expression of p62 was decreased in the BS-KS intervention group.The protein expression of PI3 K,p-Akt,and p-m TOR was also reduced.BS-KS also inhibited the m RNA expression of i NOS and CD86 on M2 macrophage,but promoted the expression of CD206 and Arg-1 on M2 macrophage.With respect to the regulation of inflammatory factors,BS-KS could inhibit the secretion of pro-inflammatory TNF-αand promote the secretion of anti-inflammatory IL-10.It also inhibited the protein expression of IKB-αand NF-κB,and promoted the expression of nuclear protein PPAR-γ.CONCLUSION:We believe that BS-KS promotes macrophage autophagy by increasing the level of autophagy protein and inhibiting the PI3 K/Akt/m TOR signaling pathway.Furthermore,BS-KS seems to inhibit macrophage M1 polarization and promote M2 polarization via the PPAR gamma/NF-κB signaling pathway,thus playing an inhibitory role in atherosclerosis.展开更多
目的探讨应用补肾抗衰片配合常规西医治疗老年肾虚痰瘀型2型糖尿病肾病的临床效果。方法选取2013年4月~2016年4月我院收治的194例老年肾虚痰瘀型2型糖尿病肾病患者,按照就诊ID号奇、偶数分为观察组(n=97)与对照组(n=97),对照组患者仅接...目的探讨应用补肾抗衰片配合常规西医治疗老年肾虚痰瘀型2型糖尿病肾病的临床效果。方法选取2013年4月~2016年4月我院收治的194例老年肾虚痰瘀型2型糖尿病肾病患者,按照就诊ID号奇、偶数分为观察组(n=97)与对照组(n=97),对照组患者仅接受常规西医治疗,观察组患者在对照组的基础上接受补肾抗衰片治疗,比较两组患者治疗前后生化指标、中医症候改善情况。结果经16周治疗后,两组患者各项生化指标以及中医症候评分较治疗前均明显下降(P<0.05);观察组患者淋巴细胞群CD4+/CD8+、尿ACR、尿Alb、FBG、2 h PBG、Hb A1c均优于对照组患者,差异有统计学意义(P<0.05);观察组气短懒言、肢体麻木、排尿频多、腰背痛、脘腹胀满评分改善情况均优于对照组,差异有统计学意义(P<0.05)。结论应用补肾抗衰片配合常规西医治疗老年肾虚痰瘀型2型糖尿病肾病安全可行,可有效维持机体血糖水平稳定,调节机体免疫能力,显著改善躯体中医症候,可在临床治疗中推广应用。展开更多
基金Supported by National Natural Science Foundation of China(Effect of Bushen Kangshuai Tablet on Early Atherosclerosis from Rho/ROCK Pathway Based on Adventitia DamageNo.81173244)the Project Funding given to the Second Batch of National"Ten Thousand People Plan"Million Project Leader(No.20160621).
文摘OBJECTIVE:To investigate the efficacy of Bushen Kangshuai(BS-KS)tablet on autophagy and polarization in mouse macrophage RAW 264.7.MEYHODS:Macrophage autophagy was induced by oxidized low-density lipoprotein(100μg/m L).To detect the levels of autophagy,macrophage were transfected with double fluorescence LC3 autophagy adenovirus,then the numbers of autophagosomes and autophagic lysosomes were asessed by confocal microscopy.The autophagy related proteins expression of PI3 K,Akt,phospho-m Akt(p-Akt)and m TOR,phospho-m TOR(p-TOR),p62,microtubule-associated protein 1(LC3-Ⅱ)were determined by western blotting.The macrophage polarization model was induced by lipopolysaccharide(1μg/m L).The m RNA levels of i NOS,CD86(M1 macrophages marker molecules),and CD206,Arg-1(M2 macrophages marker molecules)were detected by real-time quantitative PCR.The concentration of cytokines TNF-αand IL-10 was determined by enzyme-linked immunosorbent assay.The protein expression of nuclear proteins PPAR-γ,NF-κB,and cytoplasmic protein IKBαwas determined by western blotting.RESULTS:The expression of the autophagy-related protein LC3-Ⅱwas increased and the expression of p62 was decreased in the BS-KS intervention group.The protein expression of PI3 K,p-Akt,and p-m TOR was also reduced.BS-KS also inhibited the m RNA expression of i NOS and CD86 on M2 macrophage,but promoted the expression of CD206 and Arg-1 on M2 macrophage.With respect to the regulation of inflammatory factors,BS-KS could inhibit the secretion of pro-inflammatory TNF-αand promote the secretion of anti-inflammatory IL-10.It also inhibited the protein expression of IKB-αand NF-κB,and promoted the expression of nuclear protein PPAR-γ.CONCLUSION:We believe that BS-KS promotes macrophage autophagy by increasing the level of autophagy protein and inhibiting the PI3 K/Akt/m TOR signaling pathway.Furthermore,BS-KS seems to inhibit macrophage M1 polarization and promote M2 polarization via the PPAR gamma/NF-κB signaling pathway,thus playing an inhibitory role in atherosclerosis.
文摘目的探讨应用补肾抗衰片配合常规西医治疗老年肾虚痰瘀型2型糖尿病肾病的临床效果。方法选取2013年4月~2016年4月我院收治的194例老年肾虚痰瘀型2型糖尿病肾病患者,按照就诊ID号奇、偶数分为观察组(n=97)与对照组(n=97),对照组患者仅接受常规西医治疗,观察组患者在对照组的基础上接受补肾抗衰片治疗,比较两组患者治疗前后生化指标、中医症候改善情况。结果经16周治疗后,两组患者各项生化指标以及中医症候评分较治疗前均明显下降(P<0.05);观察组患者淋巴细胞群CD4+/CD8+、尿ACR、尿Alb、FBG、2 h PBG、Hb A1c均优于对照组患者,差异有统计学意义(P<0.05);观察组气短懒言、肢体麻木、排尿频多、腰背痛、脘腹胀满评分改善情况均优于对照组,差异有统计学意义(P<0.05)。结论应用补肾抗衰片配合常规西医治疗老年肾虚痰瘀型2型糖尿病肾病安全可行,可有效维持机体血糖水平稳定,调节机体免疫能力,显著改善躯体中医症候,可在临床治疗中推广应用。
