大蒜素通过抑制OPN表达和NF‑κB通路活化对大鼠肾结石形成的作用研究

doi:10.19586/j.2095-2341.2021.0106

生物技术进展 ›› 2022, Vol. 12 ›› Issue (2): 305-312.DOI: 10.19586/j.2095-2341.2021.0106

大蒜素通过抑制OPN表达和NF‑κB通路活化对大鼠肾结石形成的作用研究

徐旭红(), 孙建明, 张伦中, 许良余, 曾向阳, 孙龙飞, 陈晓峰()

郴州市第一人民医院泌尿外科，湖南郴州 423000

收稿日期:2021-06-10 接受日期:2021-10-08 出版日期:2022-03-25 发布日期:2022-03-25
通讯作者: 陈晓峰
作者简介:徐旭红 E-mail：houhhm@163.com；

Study on the Effect of Allicin on Rat Kidney Stone Formation by Inhibiting OPN Expression and Activation of NF‑κB Pathway

Xuhong XU(), Jianming SUN, Lunzhong ZHANG, Liangyu XU, Xiangyang ZENG, Longfei SUN, Xiaofeng CHEN()

Department of Urology，the First People's Hospital of Chenzhou City，Hunan Chenzhou 423000，China

Received:2021-06-10 Accepted:2021-10-08 Online:2022-03-25 Published:2022-03-25
Contact: Xiaofeng CHEN

摘要/Abstract

摘要：

大蒜素可改善草酸诱导的肾小管上皮细胞损伤，以肾结石大鼠为研究对象，探讨大蒜素对肾结石大鼠的作用及其可能的机制。采用1%乙二醇+2%氯化铵混合液灌胃造模（空白组除外），分别灌胃大蒜素7.5 mg·kg^-1（低剂量大蒜素组）、15 mg·kg^-1（高剂量大蒜素组）、胃枸橼酸氢钾钠颗粒0.6 g·kg^-1（阳性对照组），其余组灌胃0.9%氯化钠溶液（空白组），检测各组大鼠与肾结石疾病相关的指标。与空白组相比，模型组大鼠肾指数、肌酐（creatinine，Cr）、血清尿素氮（blood urea nitrogen，BUN）水平和天冬氨酸转氨酶（aspartate aminotransferase，AST）、谷丙转氨酶（alanine aminotransferase，ALT）活性及24 h尿量、尿液中草酸、钙和磷含量显著升高（P<0.05），草酸钙结晶评分显著升高（P<0.05），镁含量显著降低（P<0.05），骨桥蛋白（osteopontin，OPN）表达显著升高（P<0.05），核因子κB（nuclear factor-κB，NF-κB）通路活化；与模型组相比，低剂量大蒜素组、高剂量大蒜素组和阳性对照组大鼠肾指数、Cr、BUN水平和AST、ALT活性、24 h尿量、尿液中草酸、钙和磷含量显著降低（P<0.05），草酸钙结晶评分显著降低（P<0.05），镁含量显著升高（P<0.05），OPN表达显著降低（P<0.05），NF?κB通路被抑制。结果表明，大蒜素通过改善大鼠肾功能指标、抑制骨桥蛋白表达和NF?κB通路活化进而抑制肾结石形成。

关键词: 大蒜素, 肾结石, 骨桥蛋白, NF?κB通路

Abstract:

Allicin can improve the injury of renal tubular epithelial cells induced by oxalic acid， this study took rats with kidney stones as the research object to explore the effect of allicin on rats with kidney stones and its possible mechanism. 1% ethylene glycol+2% ammonium chloride method to prepare a model of rats with kidney stones （except blank group）. Allicin 7.5 mg·kg^-1 （low allicin dose group）， 15 mg·kg^-1 （high allicin dose group）， potassium sodium hydrogen citrate 0.6 g·kg^-1 （positive control group）， and 0.9% sodium chloride solution for other groups （blank group） were gavaged respectively. The indexes related to kidney stone disease in each group were detected. Compared with the blank group， the model group had significantly higher renal index， Cr， BUN levels， aspartate aminotransferase （AST）， alanine aminotransferase （ALT） activity， 24 h urine output， urine oxalic acid， calcium and phosphorus content （P<0.05）， and the calcium oxalate crystallization score was significantly increased （P<0.05）， magnesium content was significantly decreased （P<0.05）， osteopontin （OPN） expression was significantly increased （P<0.05）， nuclear factor?κB（NF?κB） pathway was activated. Compared with the model group， the renal index， Cr， BUN levels and AST， ALT activity， 24 h urine output， urine oxalic acid， calcium and phosphorus content in the low allicin dose group， high dose group and the positive control group were significantly reduced （P<0.05）， and calcium oxalate crystal score was significantly reduced （P<0.05）， the magnesium content was significantly increased （P<0.05）， OPN expression was significantly reduced （P<0.05）， and the NF?κB pathway was inhibited. Allicin can inhibit the formation of kidney stones by improving the renal function indexes of rats， inhibiting the expression of osteopontin and the activation of NF?κB pathway.

Key words: allicin, kidney stones, osteopontin, NF?κB pathway

中图分类号:

R692.4

徐旭红, 孙建明, 张伦中, 许良余, 曾向阳, 孙龙飞, 陈晓峰. 大蒜素通过抑制OPN表达和NF‑κB通路活化对大鼠肾结石形成的作用研究[J]. 生物技术进展, 2022, 12(2): 305-312.

Xuhong XU, Jianming SUN, Lunzhong ZHANG, Liangyu XU, Xiangyang ZENG, Longfei SUN, Xiaofeng CHEN. Study on the Effect of Allicin on Rat Kidney Stone Formation by Inhibiting OPN Expression and Activation of NF‑κB Pathway[J]. Current Biotechnology, 2022, 12(2): 305-312.

图/表 12

表1 5组大鼠肾指数、饮水量和进食量比较（x±s）

Table 1 Comparison of renal index, water intake and food intake among five groups (x±s)

组别	肾指数/（mg·g^-1）	饮水量/mL	进食量/g
空白组	9.32±0.32	33.48±4.05	26.56±2.78
模型组	16.29±0.42^*	18.91±4.27^*	13.40±2.45^*
阳性对照组	10.74±0.49^#	32.98±3.52^#	26.49±2.86^#
低剂量大蒜素组	12.69±0.62^#	27.68±2.25^#	20.49±3.79^#
高剂量大蒜素组	10.21±0.40^#	35.12±3.54^#	26.76±2.60^#

表2 5组大鼠Cr、BUN、AST和ALT的比较（x±s）

Table 2 Comparison of Cr, BUN, AST and ALT among five groups (x±s)

组别	Cr/（μmol·L^-1）	BUN/（mmol·L^-1）	AST/（U·L^-1）	ALT/（U·L^-1）
空白组	26.29±4.00	7.54±0.71	35.04±3.73	21.32±3.99
模型组	83.43±4.64^*	17.25±1.53^*	80.45±2.87^*	82.96±6.55^*
阳性对照组	36.29±2.99^#	8.96±0.75^#	43.93±3.29^#	35.02±4.11^#
大蒜素低剂量组	45.25±3.69^#	12.46±1.15^#	47.94±4.44^#	44.65±2.68^#
大蒜素高剂量组	34.29±2.68^#	9.37±1.02^#	43.94±3.86^#	35.46±3.88^#

图1 各组大鼠尿液中草酸钙结晶的观察注：图中放大倍数为400倍。

Fig.1 Observation of calcium oxalate crystals in urine of rats in each group

表3 5组大鼠尿量和草酸含量比较（x±s）

Table 3 Comparison of urine volume and oxalic acid content among five groups (x±s)

组别	24 h尿量/mL	草酸/（ng·L^-1）
空白组	11.70±1.33	21.28±2.42
模型组	18.71±2.53^*	39.36±2.53^*
阳性对照组	12.51±1.91^#	25.55±2.51^#
低剂量大蒜素组	13.53±1.62^#	31.75±3.23^#
高剂量大蒜素组	12.41±1.37^#	24.23±2.25^#

表4 5组大鼠尿液中钙、镁和磷含量比较（x±s）

Table 4 Comparison of calcium, magnesium and phosphorus contents in urine among five groups (x±s)

组别	钙含量/（mmol·L^-1）	镁含量/（mmol·L^-1）	磷含量/（mmol·L^-1）
空白组	2.12±0.10	3.57±0.86	0.32±0.03
模型组	3.78±0.55^*	0.74±0.08^*	1.88±0.43^*
阳性对照组	2.43±0.41^#	2.48±0.46^#	0.62±0.10^#
大蒜素低剂量组	3.08±0.32^#	1.84±0.34^#	1.07±0.09^#
大蒜素高剂量组	2.42±0.40^#	2.60±0.44^#	0.68±0.07^#

图2 HE染色检测各组大鼠肾脏草酸钙结晶注：图中放大倍数为400倍。

Fig. 2 HE staining for calcium oxalate crystals in rats kidney in each group

表5 5组大鼠草酸钙结晶评分比较（x±s）

Table 5 Calcium oxalate crystal score of rats among five groups (x±s)

组别	结晶评分/分
空白组	1.00±0.00
模型组	3.50±0.50^*
阳性对照组	1.95±0.23^#
大蒜素低剂量组	2.47±0.22^#
大蒜素高剂量组	1.92±0.26^#

图3 免疫组化检测各组大鼠肾组织中OPN表达注：图中放大倍数为400倍。

Fig. 3 Immunohistochemical detection of OPN expression in renal tissues of rats in each group

图4 Western blot检测各组大鼠肾组织中OPN表达注：1—空白组；2—模型组；3—阳性对照组；4—大蒜素低剂量组；5—大蒜素高剂量组。

Fig. 4 Western blot detection of OPN expression in kidney tissues of rats in each group

表6 5组大鼠肾组织中OPN mRNA和蛋白表达比较（x±s）

Table 6 Comparison of OPN mRNA and protein expression in kidney tissues among five groups (x±s)

组别	OPN mRNA表达	OPN蛋白表达
空白组	1.00±0.05	0.14±0.01
模型组	3.35±0.19^*	0.83±0.07^*
阳性对照组	1.43±0.09^#	0.42±0.05^#
大蒜素低剂量组	2.40±0.23^#	0.61±0.04^#
大蒜素高剂量组	1.52±0.12^#	0.44±0.04^#

图5 Western blot检测各组大鼠肾组织中NF-κb通路的表达注：1—空白组；2—模型组；3—阳性对照组；4—大蒜素低剂量组；5—大蒜素高剂量组。

Fig.5 Western blot results of NF-κB pathway in renal tissues of rats in each group

表7 5组大鼠肾组织中p-NF-κB/NF-κB表达比较（x±s）

Table 7 Comparison of expression of p-NF-κB/NF-κB in renal tissues of rats among five groups (x±s)

组别	pNFκB/NFκB表达
空白组	0.35±0.03
模型组	2.53±0.28^*
阳性对照组	1.17±0.13^#
大蒜素低剂量组	1.88±0.14^#
大蒜素高剂量组	1.10±0.13^#

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大蒜素通过抑制OPN表达和NF‑κB通路活化对大鼠肾结石形成的作用研究

Study on the Effect of Allicin on Rat Kidney Stone Formation by Inhibiting OPN Expression and Activation of NF‑κB Pathway

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