人参皂苷Rg1对肾间质纤维化大鼠肾组织肝细胞生长因子及转化生长因子-β_1的影响
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摘要
目的探讨人参皂苷Rg1对肾间质纤维化(renal interstitial fibrosis, RIF)大鼠肾组织肝细胞生长因子(hepatocyte growth factor, HGF)、转化生长因子-β_1(transforming growth factor-β_1, TGF-β_1)的调节作用。方法 60只SD大鼠随机分为观察组、对照组各30只。2组大鼠均应用腺嘌呤混悬液150 mg/(kg·d)灌胃,连续17周,制备RIF模型。造模成功后观察组腹腔注射人参皂苷Rg1 20 mL/(kg·d),连续17周;对照组腹腔注射等量生理盐水。分别于造模成功、干预17周时留取24 h尿量;采集腹主动脉血测定血肌酐水平;各组均处死15只大鼠取肾组织行病理检查,评估肾小管、肾间质损伤程度,免疫组织化学法检测肾间质Ⅰ、Ⅲ、Ⅳ型胶原及纤维连接蛋白(fibronectin, FN)、HGF、TGF-β_1表达。结果 2组造模成功时血肌酐、24 h尿量比较差异无统计学意义(P>0.05);干预17周2组血肌酐均较造模成功时降低(P<0.05)、24 h尿量较造模成功时减少(P<0.05),且观察组变化较对照组明显(P<0.05);观察组和对照组造模成功时肾间质Ⅰ型胶原(20.32±3.35、20.83±5.73)、Ⅲ型胶原(10.36±3.37、11.25±2.62)、Ⅳ型胶原(12.40±5.13、13.86±2.23)、FN(9.39±1.35、9.95±2.25)、HGF(6.58±2.37、6.25±1.15)、TGF-β_1(28.83±3.36、28.15±5.29)蛋白积分吸光度(optical density, OD)值比较差异无统计学意义(P>0.05),干预17周Ⅰ型胶原(8.83±1.54、13.22±1.67)、Ⅲ型胶原(3.63±0.62、6.58±1.49)、Ⅳ型胶原(5.50±1.24、10.65±0.94)、FN(4.72±0.63、7.05±0.41)、TGF-β_1(13.93±0.62、23.58±0.49)蛋白积分OD值均较造模成功时降低(P<0.05),HGF蛋白积分OD值(17.83±0.94、8.90±0.87)较造模成功时增高(P<0.05),且观察组变化较对照组明显(P<0.05);对照组和观察组造模成功时肾小管损伤评分[(2.89±0.58)、(2.80±0.26)分]、肾间质损伤评分[(2.95±0.36)、(2.98±0.31)分]比较差异无统计学意义(P>0.05),干预17周肾小管损伤评分[(2.33±0.16)、(0.83±0.27)分]、肾间质损伤评分[(2.56±0.33)、(0.73±0.15)分]均较造模成功时降低(P<0.05),且观察组变化较对照组明显(P<0.05)。结论人参皂苷Rg1可通过上调肾间质HGF水平,抑制TGF-β_1表达来延缓RIF进展,发挥肾组织保护作用。
Objective To investigate the role of ginsenoside Rg1 in regulating hepatocyte growth factor(HGF) and transforming growth factor-β_1(TGF-β_1) in the kidney of rats with renal interstitial fibrosis(RIF). Methods Sixty SD rats were randomly divided into observation group and control group, with 30 rats in each group. Both groups received lavage with adenine suspension 150 mg/(kg·d) for 17 weeks to prepare RIF models. After modeling, observation group was intraperitoneally injected with ginsenoside Rg1 20 mL/(kg·d) for 17 weeks, and control group was intraperitoneally injected with the same volume of normal saline. The 24 h urine volumes and the serum creatinine levels in the abdominal aorta were measured in both groups after modeling and 17-week intervention, respectively. Fifteen rats were sacrificed for pathological examination to evaluate the degree of renal tubular and renal interstitial damage. Immunohistochemistry was used to detect the expressions of typeⅠ,Ⅲ and Ⅳ collagen,fibronectin(FN),HGF and TGF-β1 in renal interstitial cells.Results There were no significant differences in serum creatinine and 24 hurine volume after modeling between two groups(P>0.05).The serum creatinine was significantly lower and 24 hurine volume was significantly less after17-week intervention than that after modeling(P<0.05),which was more obvious in observation group(P<0.05).There were no significant differences in the optical density(OD)values of renal interstitial typeⅠcollagen(20.32±3.35 vs.20.83±5.73),typeⅢ collagen(10.36±3.37 vs.11.25±2.62),type Ⅳ collagen(12.40±5.13 vs.13.86±2.23),FN(9.39±1.35 vs.9.95±2.25),HGF(6.58±2.37 vs.6.25±1.15)and TGF-β1(28.83±3.36 vs.28.15±5.29)after modeling between observation group and control group(P>0.05).The OD values of type Ⅰ collagen(8.83±1.54,13.22±1.67),typeⅢcollagen(3.63±0.62,6.58±1.49),typeⅣ collagen(5.50±1.24,10.65±0.94),FN(4.72±0.63,7.05±0.41),and TGF-β1(13.93±0.62,23.58±0.49)protein after 17-week intervention were significantly lower than those after modeling in both groups(P<0.05),and the OD values of HGF protein(17.83±0.94,8.90±0.87)were significantly higher than those after modeling(P<0.05),which was more obvious in observation group(P<0.05).There were no significant differences in renal tubular injury scores(2.89±0.58 vs.2.80±0.26)and renal interstitial injury scores(2.95±0.36 vs.2.98±0.31)after modeling between control group and observation group(P>0.05).The renal tubular injury scores(2.33±0.16,0.83±0.27)and renal interstitial injury scores(2.56±0.33,0.73±0.15)after 17-week intervention were significantly lower than those after modeling in control group and observation group(P<0.05),which was more obvious in observation group(P<0.05).Conclusion Ginsenoside Rg1 can slow the progression of RIF to protect the renal tissue by up-regulating renal interstitial HGF levels and inhibiting TGF-β1 expression.
Objective To investigate the role of ginsenoside Rg1 in regulating hepatocyte growth factor(HGF) and transforming growth factor-β_1(TGF-β_1) in the kidney of rats with renal interstitial fibrosis(RIF). Methods Sixty SD rats were randomly divided into observation group and control group, with 30 rats in each group. Both groups received lavage with adenine suspension 150 mg/(kg·d) for 17 weeks to prepare RIF models. After modeling, observation group was intraperitoneally injected with ginsenoside Rg1 20 mL/(kg·d) for 17 weeks, and control group was intraperitoneally injected with the same volume of normal saline. The 24 h urine volumes and the serum creatinine levels in the abdominal aorta were measured in both groups after modeling and 17-week intervention, respectively. Fifteen rats were sacrificed for pathological examination to evaluate the degree of renal tubular and renal interstitial damage. Immunohistochemistry was used to detect the expressions of typeⅠ,Ⅲ and Ⅳ collagen,fibronectin(FN),HGF and TGF-β1 in renal interstitial cells.Results There were no significant differences in serum creatinine and 24 hurine volume after modeling between two groups(P>0.05).The serum creatinine was significantly lower and 24 hurine volume was significantly less after17-week intervention than that after modeling(P<0.05),which was more obvious in observation group(P<0.05).There were no significant differences in the optical density(OD)values of renal interstitial typeⅠcollagen(20.32±3.35 vs.20.83±5.73),typeⅢ collagen(10.36±3.37 vs.11.25±2.62),type Ⅳ collagen(12.40±5.13 vs.13.86±2.23),FN(9.39±1.35 vs.9.95±2.25),HGF(6.58±2.37 vs.6.25±1.15)and TGF-β1(28.83±3.36 vs.28.15±5.29)after modeling between observation group and control group(P>0.05).The OD values of type Ⅰ collagen(8.83±1.54,13.22±1.67),typeⅢcollagen(3.63±0.62,6.58±1.49),typeⅣ collagen(5.50±1.24,10.65±0.94),FN(4.72±0.63,7.05±0.41),and TGF-β1(13.93±0.62,23.58±0.49)protein after 17-week intervention were significantly lower than those after modeling in both groups(P<0.05),and the OD values of HGF protein(17.83±0.94,8.90±0.87)were significantly higher than those after modeling(P<0.05),which was more obvious in observation group(P<0.05).There were no significant differences in renal tubular injury scores(2.89±0.58 vs.2.80±0.26)and renal interstitial injury scores(2.95±0.36 vs.2.98±0.31)after modeling between control group and observation group(P>0.05).The renal tubular injury scores(2.33±0.16,0.83±0.27)and renal interstitial injury scores(2.56±0.33,0.73±0.15)after 17-week intervention were significantly lower than those after modeling in control group and observation group(P<0.05),which was more obvious in observation group(P<0.05).Conclusion Ginsenoside Rg1 can slow the progression of RIF to protect the renal tissue by up-regulating renal interstitial HGF levels and inhibiting TGF-β1 expression.
引文
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[13] 瞿丛新,郭小军,李泉源,等.乳腺癌组织中EphA2、SIAH2、β-catenin的表达量及其与肿瘤恶性程度的相关性[J].临床和实验医学杂志,2016,15(9):844-847.
[14] 李娅妮,梁柳丹,陈燕琼,等.旁路信号通路激活介导的EML4-ALK融合基因阳性肺癌细胞株H3122对alectinib继发耐药的研究[J].中国病理生理杂志,2017,33(5):769-775.
[15] 艾麦提·牙森,金鑫,陈梓昕,等.TGF-β1信号介导的肝星状细胞促进胚胎肝前体细胞向胆管细胞方向分化[J].第三军医大学学报,2018,40(9):746-752.
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[2] DICARA D M,CHIRGADZE D Y,POPE A R,et al.Characterization and structural determination of a new anti-MET function-blocking antibody with binding epitope distinct from the ligand binding domain[J].Sci Rep,2017,7(1):9000.
[3] JIANG Q F,HUANG M Y,WU K Y,et al.Intervention effects of atorvastatin combined with panax notoginseng saponins on rats with atherosclerosis complicated with hepatic injury[J].Pharmacogn Mag,2017,13(51):430-438.
[4] 刘凌汐,于洋.雷公藤多苷片联合复方α-酮酸片治疗糖尿病肾病的临床观察[J].中国药房,2017,28(33):4654-4657.
[5] 陈建泉,张建成,陈林,等.肝细胞生长因子/转化生长因子-β1对人心房成纤维细胞增殖的作用[J].中华老年多器官疾病杂志,2017,16(11):812-817.
[6] ZHAN H,HUANG F,MA W,et al.Protective effect of ginsenoside Rg1 on bleomycin-induced pulmonary fibrosis in rats:involvement of caveolin-1 and TGF-β1 signal pathway[J].Biol Pharm Bull,2016,39(8):1284-1292.
[7] 王红月,张洋,刘丽,等.PCI-neo-肝细胞生长因子对脂多糖刺激的大鼠肾小球系膜细胞α-平滑肌肌动蛋白表达的影响[J].中国实验诊断学,2016,20(8):1264-1267.
[8] 余晓红,吕宏迪,杨旸,等.转化生长因子-β1和Smad7信号通路与大鼠肝纤维化的相关性[J].中华实用诊断与治疗杂志,2015,29(7):648-650.
[9] 宋菲,原博.脂肪干细胞来源的条件培养基对TGF-β1共同干预下的真皮成纤维细胞致纤维化能力的影响[J].上海交通大学学报(医学版),2017,37(5):588-594.
[10] MAN S,QIU P,LI J,et al.Global metabolic profiling for the study of Rhizoma Paridis saponins-induced hepatotoxicity in rats[J].Environ Toxicol,2017,32(1):99-108.
[11] ZANONI P,KHETARPAL S A,LARACH D B,et al.Rare variant in scavenger receptor B1 raises HDL cholesterol and increases risk of coronary heart disease[J].Science,2016,351(6278):1166-1171.
[12] 张新志,黄迪,吴锋,等.TGF-β1/p38MAPK通路对肾间质纤维化影响及抗纤灵冲剂干预机制的实验研究[J].中华中医药杂志,2011,26(2):245-248.
[13] 瞿丛新,郭小军,李泉源,等.乳腺癌组织中EphA2、SIAH2、β-catenin的表达量及其与肿瘤恶性程度的相关性[J].临床和实验医学杂志,2016,15(9):844-847.
[14] 李娅妮,梁柳丹,陈燕琼,等.旁路信号通路激活介导的EML4-ALK融合基因阳性肺癌细胞株H3122对alectinib继发耐药的研究[J].中国病理生理杂志,2017,33(5):769-775.
[15] 艾麦提·牙森,金鑫,陈梓昕,等.TGF-β1信号介导的肝星状细胞促进胚胎肝前体细胞向胆管细胞方向分化[J].第三军医大学学报,2018,40(9):746-752.
[16] ZYWIEL M G,CHERIAN J J,BANERJEE S,et al.Systemic cobalt toxicity from total hip arthroplasties:review of a rare condition Part 2.measurement,risk factors,and step-wise approach to treatment[J].Bone Joint J,2016,98B(1):14-20.