芪苈强心颗粒对心肾综合征大鼠肾组织细胞凋亡的影响
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摘要
目的:探讨芪苈强心颗粒对心肾综合征(cardiorenal syndrome,CRS)模型大鼠肾组织细胞凋亡的影响及可能作用机制。方法:采用左冠状动脉前降支结扎结合肾脏急性缺血再灌注损伤制备CRS模型,根据实验需要分为6组:2周假手术(2w sham)组、2周模型(2w CRS)组、2周药物(2w CRS-Q)组、4周假手术(4w sham)组、4周模型(4w CRS)组和4周药物(4w CRS-Q)组,2周和4周药物组分别给予芪苈强心颗粒(4 g·kg~(-1)·d~(-1))灌胃治疗2周和4周。ELISA法检测血清胱抑素C(Cys-C)、血浆血管紧张素Ⅱ(Ang Ⅱ)、尿中性粒细胞明胶酶相关脂质运载蛋白(NGAL)和尿微量白蛋白(UMA)含量;肌氨酸氧化酶法检测血清肌酐(Cre)含量;HE染色观察大鼠肾组织病理学变化;RT-qPCR法和Western blot检测大鼠肾组织中Ang Ⅱ、Bcl-2和Bax的mRNA和蛋白表达水平;TUNEL染色检测肾组织细胞凋亡。结果:与sham组比较,2w CRS和4w CRS组大鼠血清Cys-C、血清Cre、血浆Ang Ⅱ、UMA和尿NGAL含量均显著升高(P<0.05),肾组织中Bax和Ang Ⅱ的mRNA和蛋白表达水平均显著上调(P<0.05),Bcl-2的mRNA和蛋白表达水平均显著下调(P<0.05),肾组织损伤均严重,肾组织细胞凋亡均明显;与CRS组比较,2w CRS-Q和4w CRS-Q组大鼠血清Cys-C、血清Cre、血浆Ang Ⅱ、尿NGAL和UMA含量均显著降低(P<0.05),肾组织中Bax和Ang Ⅱ的mRNA和蛋白表达水平均显著下调(P<0.05),Bcl-2的mRNA和蛋白表达水平均显著上调(P<0.05),肾组织损伤均有所改善,肾组织细胞凋亡率均显著降低(P<0.05)。结论:芪苈强心颗粒可抑制CRS大鼠肾组织细胞凋亡,改善肾功能,其机制可能与抑制Ang Ⅱ的表达有关。
AIM: To investigate the effect of Qiliqiangxin granule on the apoptosis of renal tissues in rats with cardiorenal syndrome(CRS) and its possible mechanism. METHODS: A rat model of CRS was established by ligation of the left anterior descending coronary artery and acute renal ischemia/reperfusion injury. After operation, the rats were divided into 6 groups: 2-week sham operation(2 w sham) group, 2-week model(2 w CRS) group, 2-week drug(2 w CRS-Q) group, 4-week sham operation(4 w sham) group, 4-week model(4 w CRS) group and 4-week drug(4 w CRS-Q) group. The rats in 2 w CRS-Q group and 4 w CRS-Q group were given Qiliqiangxin granule(4 g·kg~(-1)·d~(-1)) by gavage for 2 weeks and 4 weeks, respectively. The levels of serum cystatin C(Cys-C), plasma angiotensin Ⅱ(Ang Ⅱ), urine neutrophil gelatinase-associated lipocalin(NGAL) and urine microalbumin(UMA) were measured by ELISA. The serum level of creatinine(Cre) was detected by sarcosine oxidase method. The renal histopathological changes were observed by HE staining. The mRNA and protein expression levels of Ang Ⅱ, Bcl-2 and Bax were evaluated by RT-qPCR and Western blot, respectively. The apoptosis rate of renal cells was assessed by TUNEL staining. RESULTS: The levels of serum Cys-C, serum Cre, plasma Ang Ⅱ, urine NGAL and UMA were significantly increased in 2 w CRS group and 4 w CRS group compared with 2 w sham group and 4 w sham group after modeling(P<0.05). The mRNA and protein expression levels of Bax and Ang Ⅱ in the renal tissues of CRS rats were significantly up-regulated(P<0.05), while Bcl-2 was significantly down-regulated(P<0.05) compared with 2 w sham group and 4 w sham group. Compared with 2 w sham group and 4 w sham group, the damage of renal tissues in 2 w CRS and 4 w CRS group was severe, and the apoptotic rates of renal cells were significantly increased. Compared with 2 w CRS group and 4 w CRS group, Qiliqiangxin granule greatly decreased the levels of Cys-C, Cre, Ang Ⅱ, NGAL and UMA, down-regulated the mRNA and protein expression levels of Bax and Ang Ⅱ in the renal tissues, and up-regulated the expression of Bcl-2 at mRNA and protein levels at 2 and 4 weeks. In addition, Qiliqiangxin granule also greatly attenuated the damage and apoptosis of the renal tissues. CONCLUSION: Qiliqiangxin granule significantly inhibits the apoptosis of renal tissues and improves the renal function of CRS rats, and its mechanism may be related to the inhibition of Ang Ⅱ expression.
AIM: To investigate the effect of Qiliqiangxin granule on the apoptosis of renal tissues in rats with cardiorenal syndrome(CRS) and its possible mechanism. METHODS: A rat model of CRS was established by ligation of the left anterior descending coronary artery and acute renal ischemia/reperfusion injury. After operation, the rats were divided into 6 groups: 2-week sham operation(2 w sham) group, 2-week model(2 w CRS) group, 2-week drug(2 w CRS-Q) group, 4-week sham operation(4 w sham) group, 4-week model(4 w CRS) group and 4-week drug(4 w CRS-Q) group. The rats in 2 w CRS-Q group and 4 w CRS-Q group were given Qiliqiangxin granule(4 g·kg~(-1)·d~(-1)) by gavage for 2 weeks and 4 weeks, respectively. The levels of serum cystatin C(Cys-C), plasma angiotensin Ⅱ(Ang Ⅱ), urine neutrophil gelatinase-associated lipocalin(NGAL) and urine microalbumin(UMA) were measured by ELISA. The serum level of creatinine(Cre) was detected by sarcosine oxidase method. The renal histopathological changes were observed by HE staining. The mRNA and protein expression levels of Ang Ⅱ, Bcl-2 and Bax were evaluated by RT-qPCR and Western blot, respectively. The apoptosis rate of renal cells was assessed by TUNEL staining. RESULTS: The levels of serum Cys-C, serum Cre, plasma Ang Ⅱ, urine NGAL and UMA were significantly increased in 2 w CRS group and 4 w CRS group compared with 2 w sham group and 4 w sham group after modeling(P<0.05). The mRNA and protein expression levels of Bax and Ang Ⅱ in the renal tissues of CRS rats were significantly up-regulated(P<0.05), while Bcl-2 was significantly down-regulated(P<0.05) compared with 2 w sham group and 4 w sham group. Compared with 2 w sham group and 4 w sham group, the damage of renal tissues in 2 w CRS and 4 w CRS group was severe, and the apoptotic rates of renal cells were significantly increased. Compared with 2 w CRS group and 4 w CRS group, Qiliqiangxin granule greatly decreased the levels of Cys-C, Cre, Ang Ⅱ, NGAL and UMA, down-regulated the mRNA and protein expression levels of Bax and Ang Ⅱ in the renal tissues, and up-regulated the expression of Bcl-2 at mRNA and protein levels at 2 and 4 weeks. In addition, Qiliqiangxin granule also greatly attenuated the damage and apoptosis of the renal tissues. CONCLUSION: Qiliqiangxin granule significantly inhibits the apoptosis of renal tissues and improves the renal function of CRS rats, and its mechanism may be related to the inhibition of Ang Ⅱ expression.
引文
[1] Bongartz LG, Braam B, Gaillard CA, et al. Target organ cross talk in cardiorenal syndrome: animal models[J]. Am J Physiol Renal Physiol, 2012, 303(9):F1253-F1263.
[2] 曾爱英, 马青变. 心肾综合征诊治进展[J]. 心血管病学进展, 2017, 38(2):214-218.
[3] 金敬顺, 张霞, 马红微. 芪苈强心胶囊对老年慢性心力衰竭合并早期肾功能损害患者凝血功能及内皮细胞功能的影响[J]. 现代中西医结合杂志, 2016, 25(35):3939-3941.
[4] 卢金萍, 李夏, 陈玲, 等. 芪苈强心胶囊治疗老年慢性心肾综合征的临床研究[J]. 世界中医药, 2013, 8(12):1496-1498.
[5] Ronco C, Haapio M, House AA, et al. Cardiorenal syndrome[J]. Heart Fail Clin, 2014, 52(19):251-280.
[6] Kingma JG, Simard D, Rouleau JR, et al. The physiopathology of cardiorenal syndrome: a review of the potential contributions of inflammation[J]. J Cardiovasc Dev Dis, 2017, 4(4):21-28.
[7] Desir J, Neugarten J, Melamed M, et al. Glomerular size reduction associated with severe cardiorenal syndrome[J]. Clin Nephrol, 2016, 85(3):159-164.
[8] 方依琳. 芪苈强心胶囊对急性心肾综合征大鼠肾动脉血流、氧化应激及炎性因子影响[D].郑州: 河南中医学院,2015.
[9] 申素琴, 申玉敏. 芪苈强心胶囊对慢性心力衰竭患者心功能、神经内分泌及细胞因子的影响[J]. 河北中医, 2015, 37(9):1388-1390.
[10] Brisco MA, Testani JM. Novel renal biomarkers to assess cardiorenal syndrome[J]. Curr Heart Fail Rep, 2014, 11(4):485-499.
[11] Ogawaakiyama A, Sugiyama H, Kitagawa M, et al. Serum cystatin C is an independent biomarker associated with the renal resistive index in patients with chronic kidney disease[J]. PLoS One, 2018, 13(3):e0193695.
[12] 张翩, 王玲, 何奔, 等. 生物标志物NGAL、NAG、胱抑素C诊断对比剂急性肾损伤的价值[J]. 兰州大学学报: 医学版, 2018, 44(1):7-13.
[13] 刘静, 高娟. 芪苈强心胶囊联合阿托伐他汀治疗2型心肾综合征疗效及对血清hs-CRP、D-D、ET-1的影响[J]. 现代中西医结合杂志, 2017, 26(28):3100-3103.
[14] Zhao L, Gu Q, Xiang L, et al. Curcumin inhibits apoptosis by modulating Bax/Bcl-2 expression and alleviates oxidative stress in testes of streptozotocin-induced diabetic rats[J]. Ther Clin Risk Manag, 2017, 13:1099-1105.
[15] 任冰霜, 雷艳, 黄梁浒. 肾脏缺血再灌注损伤的分子机制[J]. 实用医学杂志, 2016, 32(10):1710-1712.
[16] 肖骏, 马渝, 邓松柏, 等. 芪苈强心对心肌梗死后大鼠心肌细胞凋亡的影响[J]. 中国病理生理杂志, 2012, 28(6):1045-1050.
[17] 夏青青, 李娟娟, 郭泽云. 血管紧张素Ⅱ及其受体在脑血管疾病中作用研究进展[J]. 神经解剖学杂志, 2012, 28(1):89-92.
[18] 戴榕, 王超, 周军, 等. 硫氢化钠对慢性心力衰竭大鼠心脏功能和肾素-血管紧张素-醛固酮系统活性的影响[J]. 中国病理生理杂志, 2018, 34(2):276-280.
[19] 叶勇, 李磊, 蒋国良, 等. 芪苈强心胶囊通过抑制血管紧张素Ⅱ改善压力超负荷致小鼠心肌肥厚[J]. 中国分子心脏病学杂志, 2011, 11(6):355-361.
[20] 涂珊, 周巧玲, 唐荣, 等. 血管紧张素Ⅱ对肾小管上皮细胞凋亡的影响及冬虫夏草对其的干预作用[J]. 中南大学学报: 医学版, 2012, 37(1):67-72.
[21] 周安宇, 余凌, 李惊子, 等. 血管紧张素ⅡⅠ型受体拮抗剂和血管紧张素转换酶抑制剂的肾保护作用及其对肾内肾素-血管紧张素系统的影响[J]. 中国病理生理杂志, 2001, 17(9):843-846.
[2] 曾爱英, 马青变. 心肾综合征诊治进展[J]. 心血管病学进展, 2017, 38(2):214-218.
[3] 金敬顺, 张霞, 马红微. 芪苈强心胶囊对老年慢性心力衰竭合并早期肾功能损害患者凝血功能及内皮细胞功能的影响[J]. 现代中西医结合杂志, 2016, 25(35):3939-3941.
[4] 卢金萍, 李夏, 陈玲, 等. 芪苈强心胶囊治疗老年慢性心肾综合征的临床研究[J]. 世界中医药, 2013, 8(12):1496-1498.
[5] Ronco C, Haapio M, House AA, et al. Cardiorenal syndrome[J]. Heart Fail Clin, 2014, 52(19):251-280.
[6] Kingma JG, Simard D, Rouleau JR, et al. The physiopathology of cardiorenal syndrome: a review of the potential contributions of inflammation[J]. J Cardiovasc Dev Dis, 2017, 4(4):21-28.
[7] Desir J, Neugarten J, Melamed M, et al. Glomerular size reduction associated with severe cardiorenal syndrome[J]. Clin Nephrol, 2016, 85(3):159-164.
[8] 方依琳. 芪苈强心胶囊对急性心肾综合征大鼠肾动脉血流、氧化应激及炎性因子影响[D].郑州: 河南中医学院,2015.
[9] 申素琴, 申玉敏. 芪苈强心胶囊对慢性心力衰竭患者心功能、神经内分泌及细胞因子的影响[J]. 河北中医, 2015, 37(9):1388-1390.
[10] Brisco MA, Testani JM. Novel renal biomarkers to assess cardiorenal syndrome[J]. Curr Heart Fail Rep, 2014, 11(4):485-499.
[11] Ogawaakiyama A, Sugiyama H, Kitagawa M, et al. Serum cystatin C is an independent biomarker associated with the renal resistive index in patients with chronic kidney disease[J]. PLoS One, 2018, 13(3):e0193695.
[12] 张翩, 王玲, 何奔, 等. 生物标志物NGAL、NAG、胱抑素C诊断对比剂急性肾损伤的价值[J]. 兰州大学学报: 医学版, 2018, 44(1):7-13.
[13] 刘静, 高娟. 芪苈强心胶囊联合阿托伐他汀治疗2型心肾综合征疗效及对血清hs-CRP、D-D、ET-1的影响[J]. 现代中西医结合杂志, 2017, 26(28):3100-3103.
[14] Zhao L, Gu Q, Xiang L, et al. Curcumin inhibits apoptosis by modulating Bax/Bcl-2 expression and alleviates oxidative stress in testes of streptozotocin-induced diabetic rats[J]. Ther Clin Risk Manag, 2017, 13:1099-1105.
[15] 任冰霜, 雷艳, 黄梁浒. 肾脏缺血再灌注损伤的分子机制[J]. 实用医学杂志, 2016, 32(10):1710-1712.
[16] 肖骏, 马渝, 邓松柏, 等. 芪苈强心对心肌梗死后大鼠心肌细胞凋亡的影响[J]. 中国病理生理杂志, 2012, 28(6):1045-1050.
[17] 夏青青, 李娟娟, 郭泽云. 血管紧张素Ⅱ及其受体在脑血管疾病中作用研究进展[J]. 神经解剖学杂志, 2012, 28(1):89-92.
[18] 戴榕, 王超, 周军, 等. 硫氢化钠对慢性心力衰竭大鼠心脏功能和肾素-血管紧张素-醛固酮系统活性的影响[J]. 中国病理生理杂志, 2018, 34(2):276-280.
[19] 叶勇, 李磊, 蒋国良, 等. 芪苈强心胶囊通过抑制血管紧张素Ⅱ改善压力超负荷致小鼠心肌肥厚[J]. 中国分子心脏病学杂志, 2011, 11(6):355-361.
[20] 涂珊, 周巧玲, 唐荣, 等. 血管紧张素Ⅱ对肾小管上皮细胞凋亡的影响及冬虫夏草对其的干预作用[J]. 中南大学学报: 医学版, 2012, 37(1):67-72.
[21] 周安宇, 余凌, 李惊子, 等. 血管紧张素ⅡⅠ型受体拮抗剂和血管紧张素转换酶抑制剂的肾保护作用及其对肾内肾素-血管紧张素系统的影响[J]. 中国病理生理杂志, 2001, 17(9):843-846.