百令胶囊对狼疮性肾炎患者巨噬细胞功能的影响
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摘要
目的探究百令胶囊调控miR-127的表达对狼疮性肾炎(LN)患者巨噬细胞功能的影响。方法选取赣州市人民医院2016年1月—2017年6月收治的47例LN患者为研究对象,随机分为2组,对照组给予糖皮质激素联合环磷酰胺治疗,治疗组在对照组治疗方案的基础上给予百令胶囊治疗,连续治疗12个月后测定白蛋白(Alb)、血肌酐(SCr)、补体C3、24 h尿蛋白定量(TP)、血红细胞(RBC)以评价临床疗效。分别用百令胶囊提取液培养细胞、miR-127 mimic、miR-127 inhibitor转染细胞。流式细胞术测定巨噬细胞凋亡率与增殖能力;qPCR法检测患者外周血巨噬细胞、人巨噬细胞miR-127表达;Western Blot法检测人巨噬细胞肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)表达。结果治疗后与对照组比较,治疗组Alb、C3、TP和RBC水平降低,治疗组总有效率较高,外周血巨噬细胞miR-127表达水平大幅下降。在细胞实验中,当百令胶囊浓度<50 mg·L~(-1)时巨噬细胞凋亡率不受影响;单独加入百令胶囊提取液、miR-127inhibitor后巨噬细胞G0/G1期比例上升,吞噬能力增强,促炎因子TNF-α,IL-1β、IL-6释放减少;在百令胶囊提取液孵育的巨噬细胞中补充miR-127后,其G0/G1期比例下降,吞噬能力下降,促炎因子TNF-α、IL-1β、IL-6释放增加。结论百令胶囊可能通过下调巨噬细胞内miR-127的表达,抑制巨噬细胞增殖和吞噬功能,减少促炎因子TNF-α、IL-1β、IL-6释放功能,有效缓解狼疮性肾炎的病情。
Objective To preliminarily explore the effect of Bailing capsules on macrophage function in patients with lupus nephritis(LN)by regulating the expression of miR-127. Methods A total of 47 LN patients admitted to our hospital from January 2016 to June 2017 were selected, and randomly divided into treatment group and control group. The control group was treated with glucocorticoid combined with cyclophosphamide,and the treatment group was treated with Bailing capsules on the basis of the control group treatment. Both groups were treated for 12 months.Then clinical efficacy was evaluated according to Alb,SCr,C3,TP and RBC results. Cells were cultured with extract of Bailing capsules and transfected with miR-127 mimic and miR-127 inhibitor,respectively. The apoptosis rate and proliferation capacity of macrophages were measured by flow cytometry. The expression of miR-127 in peripheral blood macrophages and human macrophages was detected by qPCR. The expression of human macrophage TNF-α, IL-1β, and IL-6 was detected by Western Blot. Results After treatment, compared with the control group,the Alb,C3,TP and RBC were decreased,the total effective rate was higher in the treatment group,and the expression level of miR-127 in peripheral blood macrophages was significantly decreased. In cell experiments,the apoptosis rate of macrophages was not affected when the concentration of Bailing capsule was less than 50 mg·L~(-1).The proportion of macrophages in G0/G1 phase increased after the addition of Bailing capsule extract and miR-127 inhibitor,and the phagocytosis was enhanced,and the pro-inflammatory factors TNF-α,IL-1β,and IL-6 were reduced. After adding miR-127 to the macrophages which were incubated in the extract of Bailing capsules before,the proportion of G0/G1 phase decreased,the phagocytic capacity decreased,the pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 were increased. Conclusion Bailing capsule inhibit the proliferation and phagocytosis of macrophages by down-regulating miR-127,and reduce the release of inflammatory cytokines TNF-α,IL-1 and IL-6,so as to alleviate the further development of lupus nephriti.
Objective To preliminarily explore the effect of Bailing capsules on macrophage function in patients with lupus nephritis(LN)by regulating the expression of miR-127. Methods A total of 47 LN patients admitted to our hospital from January 2016 to June 2017 were selected, and randomly divided into treatment group and control group. The control group was treated with glucocorticoid combined with cyclophosphamide,and the treatment group was treated with Bailing capsules on the basis of the control group treatment. Both groups were treated for 12 months.Then clinical efficacy was evaluated according to Alb,SCr,C3,TP and RBC results. Cells were cultured with extract of Bailing capsules and transfected with miR-127 mimic and miR-127 inhibitor,respectively. The apoptosis rate and proliferation capacity of macrophages were measured by flow cytometry. The expression of miR-127 in peripheral blood macrophages and human macrophages was detected by qPCR. The expression of human macrophage TNF-α, IL-1β, and IL-6 was detected by Western Blot. Results After treatment, compared with the control group,the Alb,C3,TP and RBC were decreased,the total effective rate was higher in the treatment group,and the expression level of miR-127 in peripheral blood macrophages was significantly decreased. In cell experiments,the apoptosis rate of macrophages was not affected when the concentration of Bailing capsule was less than 50 mg·L~(-1).The proportion of macrophages in G0/G1 phase increased after the addition of Bailing capsule extract and miR-127 inhibitor,and the phagocytosis was enhanced,and the pro-inflammatory factors TNF-α,IL-1β,and IL-6 were reduced. After adding miR-127 to the macrophages which were incubated in the extract of Bailing capsules before,the proportion of G0/G1 phase decreased,the phagocytic capacity decreased,the pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 were increased. Conclusion Bailing capsule inhibit the proliferation and phagocytosis of macrophages by down-regulating miR-127,and reduce the release of inflammatory cytokines TNF-α,IL-1 and IL-6,so as to alleviate the further development of lupus nephriti.
引文
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[14]TANIGUCHI K,HIKIJI H,OKINAGA T,et al.Essential role of lysophosphatidylcholine acyltransferase 3 in the induction of macrophage polarization in PMA-Treated U937 Cells[J].J Cell Biochem,2015,116(2):2840-2848.
[15]张楠,杨洪涛.缬沙坦联合百令胶囊对IgA肾病患者尿蛋白、尿渗透压的影响及其临床安全性的研究[J].西部中医药,2016,29(10):116-118.
[16]刘春燕,陈云霞,苏俊平,等.百令胶囊对糖尿病肾病患者肾功能的影响[J].河北医药,2016,38(1):52-54.
[17]杨志海,刘冰,陆健,等.百令胶囊对狼疮性肾炎患者临床疗效和感染率影响的多中心前瞻性研究[J].黑龙江医药,2015,28(6):1205-1208.
[18]陈伟英,杨念生,尹培达,等.巨噬细胞局部浸润和增殖在狼疮性肾炎中的作用[J].中华风湿病学杂志,1998,2(2):89-91.
[19]JIN X,YAO T,ZHOU Z,et al.Advanced glycation end products enhance macrophages polarization into M1 phenotype through activating RAGE/NF-κB pathway[J].Biomed Res Int,2015,2015:732450.
[20]WANG N,LIANG H,ZEN K.Molecular mechanisms that influence the macrophage m1-m2 polarization balance[J].Front Immunol,2014,5:614.
[21]ZHOU D,HUANG C,LIN Z,et al.Macrophage polarization and function with emphasis on the evolving roles of coordinated regulation of cellular signaling pathways[J].Cell Signal,2014,26(2):192-197.
[22]DAI Y,SUI W,LAN H,et al.Comprehensive analysis of microRNA expression patterns in renal biopsies of lupus nephritis patients[J].Rheumatol Int,2009,29(7):749-754.
[23]WANG DJ,LEGESSE-MILLER A,JOHNSON EL,et al.Regulation of the let-7a-3 promoter by NF-kappaB[J].PLoS One,2012,7(2):e31240.
[24]DAI R,AHMED SA.MICRORNA,a new paradigm for understanding immunoregulation,inflammation,and autoimmune diseases[J].Transl Res,2011,157(4):163-179.
[2]孙雯,何玉甜,陈敦金.美国风湿病学会“狼疮性肾炎的筛查、治疗及管理指南”解读[J].中华产科急救电子杂志,2012,1(2):21-24.
[3]袁红.狼疮性肾炎的研究进展[J].国际检验医学杂志,2016,37(4):523-526.
[4]王世英,陈宝平,杨素霞,等.维持性血液透析肾衰竭感染患者应用百令胶囊的临床效果分析[J].中华医院感染学杂志,2016,26(10):2308-2310.
[5]闫冬梅.百令胶囊联合缬沙坦治疗慢性肾小球肾炎尿蛋白的疗效观察[J].中国急救医学,2017,37(12):116-117.
[6]刘攀,何天祎,赵汝星,等.百令胶囊对Graves病患者自身免疫的调理作用[J].山东大学学报(医学版),2017,55(9):90-95.
[7]GEISSMANN F,MANZ M G,JUNG S,et al.Development of monocytes,macrophages,and dendritic cells[J].Science,2010,327(5966):656-661.
[8]ZARJOU A,YANG S,ABRAHAM E,et al.Identification of a microRNA signature in renal fibrosis:role of miR-21[J].American Journal of Physiology,2011,301:793-801.
[9]AKAO Y,IIO A,ITOH T,et al.Microvesicle-mediated RNAmolecule delivery system using monocytes/macrophages[J].Molecular therapy:the journal of the American Society of Gene Therapy,2011,19(2):395-399.
[10]王晓阳,王广洁,张晓雪,等.百令胶囊联合他克莫司治疗狼疮性肾炎的临床研究[J].现代药物与临床,2017,32(6):1065-1069.
[11]胡赟霞,邹耀红,胥魏,等.系统性红斑狼疮国际临床合作组提出的新标准与美国风湿病学会1997修订的系统性红斑狼疮分类标准比较分析[C]//第15次全国风湿病学学术会议论文集.无锡:江苏省无锡人民医院,2010:159-159.
[12]中华医学会风湿病学分会.系统性红斑狼疮诊断及治疗指南[J].中华风湿病学杂志,2010,14(5):342-346.
[13]李向培.解读美国风湿病学会指南推进狼疮肾炎诊治的研究[J].中华风湿病学杂志,2013,17(4):217-220.
[14]TANIGUCHI K,HIKIJI H,OKINAGA T,et al.Essential role of lysophosphatidylcholine acyltransferase 3 in the induction of macrophage polarization in PMA-Treated U937 Cells[J].J Cell Biochem,2015,116(2):2840-2848.
[15]张楠,杨洪涛.缬沙坦联合百令胶囊对IgA肾病患者尿蛋白、尿渗透压的影响及其临床安全性的研究[J].西部中医药,2016,29(10):116-118.
[16]刘春燕,陈云霞,苏俊平,等.百令胶囊对糖尿病肾病患者肾功能的影响[J].河北医药,2016,38(1):52-54.
[17]杨志海,刘冰,陆健,等.百令胶囊对狼疮性肾炎患者临床疗效和感染率影响的多中心前瞻性研究[J].黑龙江医药,2015,28(6):1205-1208.
[18]陈伟英,杨念生,尹培达,等.巨噬细胞局部浸润和增殖在狼疮性肾炎中的作用[J].中华风湿病学杂志,1998,2(2):89-91.
[19]JIN X,YAO T,ZHOU Z,et al.Advanced glycation end products enhance macrophages polarization into M1 phenotype through activating RAGE/NF-κB pathway[J].Biomed Res Int,2015,2015:732450.
[20]WANG N,LIANG H,ZEN K.Molecular mechanisms that influence the macrophage m1-m2 polarization balance[J].Front Immunol,2014,5:614.
[21]ZHOU D,HUANG C,LIN Z,et al.Macrophage polarization and function with emphasis on the evolving roles of coordinated regulation of cellular signaling pathways[J].Cell Signal,2014,26(2):192-197.
[22]DAI Y,SUI W,LAN H,et al.Comprehensive analysis of microRNA expression patterns in renal biopsies of lupus nephritis patients[J].Rheumatol Int,2009,29(7):749-754.
[23]WANG DJ,LEGESSE-MILLER A,JOHNSON EL,et al.Regulation of the let-7a-3 promoter by NF-kappaB[J].PLoS One,2012,7(2):e31240.
[24]DAI R,AHMED SA.MICRORNA,a new paradigm for understanding immunoregulation,inflammation,and autoimmune diseases[J].Transl Res,2011,157(4):163-179.