摘要
目的:观察益肾活血解毒方预处理对大鼠急性肾缺血再灌注损伤(ischemia-reperfusion injury,IRI)的保护作用并探讨其机制。方法:30只雄性SD大鼠随机分为空白组、模型组及益肾活血解毒方组(14.6 g·kg^(-1)),采用经腹侧入路结扎双侧肾蒂(45 min)的方法建立急性肾IRI大鼠模型。术前7 d开始预处理给药,空白组和模型组给予生理盐水灌胃,每日1次。再灌注24 h后取材,检测血清尿素氮(blood urea nitrogen,BUN)、血清肌酐(serum creatinine,SCr)、血清胱抑素C(Cystatin C,CysC)水平;HE染色观察肾脏组织病理学变化并行肾小管损伤评分;透射电镜观察肾小管上皮细胞超微结构变化并计算病变指数;TUNEL法检测肾小管上皮细胞凋亡情况并计算凋亡指数。结果:(1)与空白组比较,模型组大鼠血清BUN、SCr、CysC水平显著升高(P<0.05);与模型组比较,益肾活血解毒方组大鼠血清BUN、SCr、CysC水平显著降低(P<0.05)。(2)HE染色:模型组大鼠肾小管管腔明显扩张,肾小管上皮细胞大量坏死,肾间质呈现水肿状态伴有大量淋巴细胞浸润;与空白组比较,模型组大鼠肾小管损伤评分显著增加(P<0.05);与模型组比较,益肾活血解毒方组大鼠肾脏组织病理形态明显改善,肾小管损伤评分显著降低(P<0.05)。(3)透射电镜:模型组大鼠肾小管损伤严重,微绒毛脱落,细胞外基质浓集,伴随大量线粒体空泡变性和内质网扩张;与空白组比较,模型组大鼠肾小管上皮细胞超微结构病变指数显著升高(P<0.05);益肾活血解毒方组大鼠肾小管上皮细胞内存在少量线粒体肿胀和空泡变性,部分内质网扩张,肾小管上皮细胞超微结构病变指数较模型组降低(P<0.05)。(4)TUNEL:与空白组比较,模型组大鼠肾小管上皮细胞凋亡指数明显升高(P<0.05);与模型组比较,益肾活血解毒方组大鼠肾小管上皮细胞凋亡指数明显降低(P<0.05)。结论:益肾活血解毒方预处理对大鼠急性肾IRI具有保护作用,其机制可能与减轻肾小管上皮细胞损伤、抑制肾小管上皮细胞凋亡有关。
Objective:To observe the protective effect of pre-treatment with Yishen Huoxue Jiedu Formula on acute renal ischemia-reperfusion injury(IRI)in rats and explore its mechanism.Method:30 male SD rats were randomly divided into a blank group,a model group,and a Yishen Huoxue Jiedu Formula group(14.6 g·kg^(-1)).The acute renal ischemia-reperfusion injury(IRI)rat model was established by ligating bilateral renal pedicle(45 minutes).Pre-treatment administration began 7 days before surgery,and the blank group and model group were given physiological saline by gavage once a day.After 24 hours of reperfusion,samples were taken and blood urea nitrogen(BUN),serum creatinine(SCr),and serum cystatin C(Cystatin C)levels were measured;HE staining was used to observe pathological changes in renal tissue and evaluate renal tubular injury score;Observation of ultrastructural changes in renal tubular epithelial cells using transmission electron microscopy and calculation of lesion index;TUNEL method was used to detect apoptosis of renal tubular epithelial cells and calculate the apoptosis index.Result:(1)Compared with the blank group,the serum levels of BUN,SCr,and CysC in the model group rats were significantly increased(P<0.05);Compared with the model group,the serum BUN,SCr,and CysC levels of rats in the Yishen Huoxue Jiedu Formula group were significantly reduced(P<0.05).(2)HE staining:The renal tubular lumen of the model group rats was significantly dilated,with a large number of necrosis of renal tubular epithelial cells and edema in the renal interstitium accompanied by a large number of lymphocyte infiltration;Compared with the blank group,the renal tubular injury score of the model group rats significantly increased(P<0.05);Compared with the model group,the pathological morphology of the kidney tissue in the Yishen Huoxue Jiedu Formula group was significantly improved,and the renal tubular injury score was significantly reduced(P<0.05).(3)Transmission electron microscopy:The renal tubules of the model group rats were severely damaged,with microvilli shedding and extracellular matrix accumulation,accompanied by a large number of mitochondrial vacuolar degeneration and endoplasmic reticulum expansion;Compared with the blank group,the ultrastructural lesion index of renal tubular epithelial cells in the model group rats was significantly increased(P<0.05);The Yishen Huoxue Jiedu Formula group of rats showed swelling and vacuolar degeneration of a few mitochondria in the renal tubular epithelial cells,partial expansion of the endoplasmic reticulum,and a decrease in the ultrastructural lesion index of renal tubular epithelial cells compared to the model group(P<0.05).(4)TUNEL:Compared with the blank group,the apoptosis index of renal tubular epithelial cells in the model group rats was significantly increased(P<0.05);Compared with the model group,the apoptosis index of renal tubular epithelial cells in the Yishen Huoxue Jiedu Formula group was significantly reduced(P<0.05).Conclusion:Pre-treatment with Yishen Huoxue Jiedu Formula has a protective effect on acute renal ischemia-reperfusion injury in rats,and its mechanism may be related to reducing damage to renal tubular epithelial cells and inhibiting apoptosis of renal tubular epithelial cells.
作者
娄丽霞
翟琼瑶
郑丰富
程丰
王锁刚
LOU Lixia;ZHAI Qiongyao;ZHENG Fengfu;CHENG Feng;WANG Suogang(The Third Affiliated Hospital to Henan University of Chinese Medicine,Zhengzhou Henan China 450008;The First Affiliated Hospital to Henan University of Chinese Medicine,Zhengzhou Henan China 450000;Henan University of Chinese Medicine,Zhengzhou Henan China 450046;Fuyang Women and Children′s Hospital,Fuyang Anhui China 236000)
出处
《中医学报》
CAS
2024年第6期1289-1294,共6页
Acta Chinese Medicine
基金
河南省中医药科学研究专项课题项目(20-21ZY2188)
河南省高等学校重点科研计划项目(18A320001)。
关键词
益肾活血解毒方
肾脏
缺血再灌注损伤
细胞凋亡
大鼠
Yishen Huoxue Jiedu Formula
kidney
ischemia reperfusion injury
cell apoptosis
rat
