肾炎康对阿霉素肾病大鼠结缔组织生长因子和纤维连结蛋白表达的影响
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摘要
目的观察肾炎康袋泡剂对阿霉素肾病大鼠的治疗作用,并研究其作用机制。方法随机选取10只大鼠为空白对照组,剩余大鼠采用一次性尾静脉注射阿霉素的方法建立阿霉素肾病模型,并随机分为肾病模型组、肾炎康组、泼尼松组,每组10只,肾病模型组予以生理盐水连续灌胃给药8周,肾炎康组和泼尼松组分别予以肾炎康袋泡剂煎液和泼尼松连续灌胃给药8周。收集末次给药后24 h尿液检测尿蛋白水平;取动脉血检测肾功能指标(血清肌酐、尿素和尿酸);取肾脏组织进行病理学染色并计算肾小球硬化指数;采用免疫组织化学染色法检测肾组织中结缔组织生长因子(connective tissue growth factor,CTGF)和纤维连结蛋白(fibronectin,FN)蛋白表达水平;采用qPCR法检测肾组织中CTGF和FN的mRNA相对表达水平。结果肾炎康干预治疗能够显著降低阿霉素肾病大鼠的血清中肌酐、尿素、尿酸水平和尿蛋白水平(P<0.05);肾炎康能够显著改善阿霉素肾病大鼠肾脏的病理结构,抑制肾纤维化水平(P<0.05),并显著降低肾病大鼠肾小球硬化指数(P<0.05)。此外,肾炎康能够显著抑制肾病大鼠肾脏组织中CTGF、FN蛋白和mRNA表达水平(P<0.05)。对于阿霉素肾病大鼠,肾炎康整体改善效果弱于泼尼松,但治疗效果仍然明显。结论肾炎康袋泡剂对阿霉素肾病大鼠有确切的治疗和改善作用,其作用可能是通过调控CTGF和FN表达进而抑制阿霉素肾病大鼠肾纤维化来实现的。
Objective To observe the therapeutic effectiveness of Shenyankang in the treatment of adriamycin-induced nephropathy in rats, and to detect the expression of CTGF and FN in renal tissue of rats. Methods Rat models of nephropathy were established by injecting adriamycin into caudal vein. The rats were divided into blank control group, nephropathy model group, Shenyankang intervention group and prednisone group. Each group contained 10 SD rats aged 6-8 weeks. Urine protein level was detected 24 h after the last administration; arterial blood was taken for renal function; kidney was taken for pathological staining; the expression of CTGF and FN protein was detected by immunohistochemistry; the relative expression of CTGF and FN mRNA was detected by qPCR. Results(1) Shenyankang intervention treatment could significantly reduce urinary protein level in nephrotic rats(P<0.05).(2) Shenyankang could significantly reduce serum creatinine, urea and uric acid levels(P<0.05).(3) Shenyankang could significantly improve the renal pathological structure of nephrotic rats, and inhibit the level of renal fibrosis(P<0.05);(4) Shenyankang could significantly reduce glomerular sclerosis index in rats with nephropathy(P<0.05);(5) Immunohistochemical results showed that Shenyankang could significantly inhibit the expression of CTGF and FN proteins in renal tissues of nephrotic rats(P<0.05).(6) The results of qPCR showed that Shenyankang could significantly inhibit the expression of CTGF and FN mRNA in renal tissues of nephrotic rats(P<0.05).(7) For nephrotic rats, the overall improvement level of Shenyankang was weaker than that of prednisone, but the therapeutic effect was still obvious. Conclusions Shenyankang capsule has a definite therapeutic effect on adriamycin-induced nephropathy in rats. Shenyankang may play an anti-adriamycin nephropathy role by regulating CTGF and FN to inhibit renal fibrosis.
Objective To observe the therapeutic effectiveness of Shenyankang in the treatment of adriamycin-induced nephropathy in rats, and to detect the expression of CTGF and FN in renal tissue of rats. Methods Rat models of nephropathy were established by injecting adriamycin into caudal vein. The rats were divided into blank control group, nephropathy model group, Shenyankang intervention group and prednisone group. Each group contained 10 SD rats aged 6-8 weeks. Urine protein level was detected 24 h after the last administration; arterial blood was taken for renal function; kidney was taken for pathological staining; the expression of CTGF and FN protein was detected by immunohistochemistry; the relative expression of CTGF and FN mRNA was detected by qPCR. Results(1) Shenyankang intervention treatment could significantly reduce urinary protein level in nephrotic rats(P<0.05).(2) Shenyankang could significantly reduce serum creatinine, urea and uric acid levels(P<0.05).(3) Shenyankang could significantly improve the renal pathological structure of nephrotic rats, and inhibit the level of renal fibrosis(P<0.05);(4) Shenyankang could significantly reduce glomerular sclerosis index in rats with nephropathy(P<0.05);(5) Immunohistochemical results showed that Shenyankang could significantly inhibit the expression of CTGF and FN proteins in renal tissues of nephrotic rats(P<0.05).(6) The results of qPCR showed that Shenyankang could significantly inhibit the expression of CTGF and FN mRNA in renal tissues of nephrotic rats(P<0.05).(7) For nephrotic rats, the overall improvement level of Shenyankang was weaker than that of prednisone, but the therapeutic effect was still obvious. Conclusions Shenyankang capsule has a definite therapeutic effect on adriamycin-induced nephropathy in rats. Shenyankang may play an anti-adriamycin nephropathy role by regulating CTGF and FN to inhibit renal fibrosis.
引文
[1] 关建国. 肾炎康配合西医常规用药治疗IgA肾病临床研究[J]. 湖北中医杂志, 2007, 29(11): 29-30.
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[4] 谭丽珍, 邢可, 王娟. 中西医结合治疗小儿肾病综合征41例疗效观察[J]. 儿科药学杂志, 2014,20(9): 23-25.
[5] 毛俐婵, 赵晓珍, 邵朝弟, 等. 肾炎康袋泡茶治疗大鼠慢性肾炎的实验研究[J]. 湖北中医学院学报, 2004, 6(3): 11-13.
[6] Raij L, Azar S, Keane W. Mesangial immune injury, hypertension, and progressive glomerular damage in Dahl rats[J]. Kidney Int, 1984, 26(2): 137-143.
[7] 吴金玉, 黄志敏, 唐宇, 等. 三七注射液对阿霉素诱导的肾纤维化大鼠补体-炎症受体系统的影响[J]. 时珍国医国药, 2018, 29(4): 796-799.
[8] 马晓红, 何立群. 从抑制肾纤维化角度研究健脾清化方对阿霉素肾病大鼠肾功能的作用与机制[J]. 中国中西医结合杂志, 2014, 34(6): 733-738.
[9] Pedrycz A, Wieczorski M, Czerny K. Histological and histochemical assessment of the effects of a single dose adriamycin on fetal rat kidney[J]. Acta Histochem, 2005, 107(3): 215-220.
[10] Toda N, Mukoyama M, Yanagita M, et al. CTGF in kidney fibrosis and glomerulonephritis[J]. Inflamm Regen, 2018, 38(1): 14.
[11] 赵雪谦, 刘云启, 杨敏. 肾纤维化相关生长因子研究新进展[J]. 临床肾脏病杂志, 2017, 17(3): 181-184.
[12] Ito Y, Aten J, Bende JR, et al. Expression of connective tissue growth factor in human renal fibrosis[J]. Kidney Int, 1998, 53(4): 853-861.
[13] Riser BL, Denichilo M, Cortes P, et al. Regulation of connective tissue growth factor activity in cultured rat mesangial cells and its expression in experimental diabetic glomerulosclerosis[J]. J Am Soc Nephrol, 2000, 11(1): 25-38.
[14] Yokoi H, Mukoyama M, Nagae T, et al. Reduction in connective tissue growth factor by antisense treatment ameliorates renal tubulointerstitial fibrosis[J]. J Am Soc Nephrol, 2004, 15(6): 1430-1440.
[15] Okada H, Kikuta T, Kobayashi T, et al. Connective tissue growth factor expressed in tubular epithelium plays a pivotal role in renal fibrogenesis[J]. J Am Soc Nephrol, 2005, 16(1): 133-143.
[16] Sakai N, Nakamura M, Lipson KE, et al. Inhibition of CTGF ameliorates peritoneal fibrosis through suppression of fibroblast and myofibroblast accumulation and angiogenesis[J]. Sci Rep, 2017, 7(1): 5392.
[17] Thiery JP, Sleeman JP. Complex networks orchestrate epithelial-mesenchymal transitions[J]. Nat Rev Mol Cell Biol, 2006, 7(2): 131-142.
[18] Nieto MA. The ins and outs of the epithelial to mesenchymal transition in health and disease[J]. Annu Rev Cell Dev Biol, 2011, 27: 347-376.
[19] 靳晶, 王全胜. 糖尿病肾病上皮细胞向间充质细胞的转分化[J]. 临床肾脏病杂志, 2017, 17(2): 68-74.
[20] Massagué J. TGFbeta in Cancer[J]. Cell, 2008, 134(2): 215-230.
[21] O'Connor JW, Riley PN, Nalluri SM, et al. Matrix rigidity mediates TGFbeta1-induced epithelial-myofibroblast transition by controlling cytoskeletal organization and MRTF-A localization[J]. J Cell Physiol, 2015, 230(8): 1829-1839.
[22] Spencer CW, Laurent F, Jeff HT, et al. Matrix stiffness drives epithelial-mesenchymal transition and tumour metastasis through a TWIST1-G3BP2 mechanotransduction pathway[J]. Nat Cell Biol, 2015, 17(5): 678-688.
[23] Pankov R. Fibronectin at a glance[J]. J Cell Sci, 2002, 115(20): 3861-3863.
[24] Bae YK, Kim A, Kim MK, et al. Fibronectin expression in carcinoma cells correlates with tumor aggressiveness and poor clinical outcome in patients with invasive breast cancer[J]. Hum Pathol, 2013, 44(10): 2028-2037.
[25] Baneyx G, Baugh L, Vogel V. Fibronectin extension and unfolding within cell matrix fibrils controlled by cytoskeletal tension[J]. Proc Natl Acad Sci USA, 2002, 99(8): 5139-5143.
[26] Griggs LA, Hassan NT, Malik RS, et al. Fibronectin fibrils regulate tgf-β1-induced epithelial-mesenchymal transition[J]. Matrix Biol, 2017, 60: 157-175.
[2] 关建国, 徐宏, 李良, 等. 肾炎康治疗慢性肾小球肾炎32例[J]. 陕西中医, 2004, 25(4): 299-301.
[3] 霍雨祥, 关建国, 尹智功. 慢性肾炎患者肾炎康治疗前后β2-M、Cr与BUN的实验观察[J]. 标记免疫分析与临床, 2003, 10(2): 119-120.
[4] 谭丽珍, 邢可, 王娟. 中西医结合治疗小儿肾病综合征41例疗效观察[J]. 儿科药学杂志, 2014,20(9): 23-25.
[5] 毛俐婵, 赵晓珍, 邵朝弟, 等. 肾炎康袋泡茶治疗大鼠慢性肾炎的实验研究[J]. 湖北中医学院学报, 2004, 6(3): 11-13.
[6] Raij L, Azar S, Keane W. Mesangial immune injury, hypertension, and progressive glomerular damage in Dahl rats[J]. Kidney Int, 1984, 26(2): 137-143.
[7] 吴金玉, 黄志敏, 唐宇, 等. 三七注射液对阿霉素诱导的肾纤维化大鼠补体-炎症受体系统的影响[J]. 时珍国医国药, 2018, 29(4): 796-799.
[8] 马晓红, 何立群. 从抑制肾纤维化角度研究健脾清化方对阿霉素肾病大鼠肾功能的作用与机制[J]. 中国中西医结合杂志, 2014, 34(6): 733-738.
[9] Pedrycz A, Wieczorski M, Czerny K. Histological and histochemical assessment of the effects of a single dose adriamycin on fetal rat kidney[J]. Acta Histochem, 2005, 107(3): 215-220.
[10] Toda N, Mukoyama M, Yanagita M, et al. CTGF in kidney fibrosis and glomerulonephritis[J]. Inflamm Regen, 2018, 38(1): 14.
[11] 赵雪谦, 刘云启, 杨敏. 肾纤维化相关生长因子研究新进展[J]. 临床肾脏病杂志, 2017, 17(3): 181-184.
[12] Ito Y, Aten J, Bende JR, et al. Expression of connective tissue growth factor in human renal fibrosis[J]. Kidney Int, 1998, 53(4): 853-861.
[13] Riser BL, Denichilo M, Cortes P, et al. Regulation of connective tissue growth factor activity in cultured rat mesangial cells and its expression in experimental diabetic glomerulosclerosis[J]. J Am Soc Nephrol, 2000, 11(1): 25-38.
[14] Yokoi H, Mukoyama M, Nagae T, et al. Reduction in connective tissue growth factor by antisense treatment ameliorates renal tubulointerstitial fibrosis[J]. J Am Soc Nephrol, 2004, 15(6): 1430-1440.
[15] Okada H, Kikuta T, Kobayashi T, et al. Connective tissue growth factor expressed in tubular epithelium plays a pivotal role in renal fibrogenesis[J]. J Am Soc Nephrol, 2005, 16(1): 133-143.
[16] Sakai N, Nakamura M, Lipson KE, et al. Inhibition of CTGF ameliorates peritoneal fibrosis through suppression of fibroblast and myofibroblast accumulation and angiogenesis[J]. Sci Rep, 2017, 7(1): 5392.
[17] Thiery JP, Sleeman JP. Complex networks orchestrate epithelial-mesenchymal transitions[J]. Nat Rev Mol Cell Biol, 2006, 7(2): 131-142.
[18] Nieto MA. The ins and outs of the epithelial to mesenchymal transition in health and disease[J]. Annu Rev Cell Dev Biol, 2011, 27: 347-376.
[19] 靳晶, 王全胜. 糖尿病肾病上皮细胞向间充质细胞的转分化[J]. 临床肾脏病杂志, 2017, 17(2): 68-74.
[20] Massagué J. TGFbeta in Cancer[J]. Cell, 2008, 134(2): 215-230.
[21] O'Connor JW, Riley PN, Nalluri SM, et al. Matrix rigidity mediates TGFbeta1-induced epithelial-myofibroblast transition by controlling cytoskeletal organization and MRTF-A localization[J]. J Cell Physiol, 2015, 230(8): 1829-1839.
[22] Spencer CW, Laurent F, Jeff HT, et al. Matrix stiffness drives epithelial-mesenchymal transition and tumour metastasis through a TWIST1-G3BP2 mechanotransduction pathway[J]. Nat Cell Biol, 2015, 17(5): 678-688.
[23] Pankov R. Fibronectin at a glance[J]. J Cell Sci, 2002, 115(20): 3861-3863.
[24] Bae YK, Kim A, Kim MK, et al. Fibronectin expression in carcinoma cells correlates with tumor aggressiveness and poor clinical outcome in patients with invasive breast cancer[J]. Hum Pathol, 2013, 44(10): 2028-2037.
[25] Baneyx G, Baugh L, Vogel V. Fibronectin extension and unfolding within cell matrix fibrils controlled by cytoskeletal tension[J]. Proc Natl Acad Sci USA, 2002, 99(8): 5139-5143.
[26] Griggs LA, Hassan NT, Malik RS, et al. Fibronectin fibrils regulate tgf-β1-induced epithelial-mesenchymal transition[J]. Matrix Biol, 2017, 60: 157-175.