Study on the improvement effect and mechanism of nobiletin on rats with acute kidney injury

doi:10.11958/20221287

Abstract

Abstract:

Objective To investigate the ameliorating effect and mechanism of nobiletin (NOB) by regulating the autophagy mediated by silent information regulator 1 (SIRT-1)/forkhead box transcription factor O3a (FOXO3a) pathway in rats with acute kidney injury (AKI). Methods A rat model of AKI was established by a one-time intraperitoneal injection of cisplatin (20 mg/kg). Sixty SPF SD male rats were randomly grouped by random number table into the control group, the AKI model group (model group), the low-dose NOB group (NOB-L group, 200 mg/kg), the high-dose NOB group (NOB-H group, 400 mg/kg) and the high-dose NOB+SIRT-1 inhibitor EX527 group (NOB-H+EX527 group, 400 mg/kg NOB+5 mg/kg EX527), with 12 rats in each group. The 24-hour urine of rats was collected after the last administration. The 24-hour urine microalbumin content, urine N-acetyl-β-D-glucosaminidase (NAG) content and urinary β2-microglobulin (β2-MG) content were detected. The serum levels of urea nitrogen (BUN) and creatinine (Scr), levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in kidney were detected by enzyme-linked immunosorbent assay (ELISA). HE staining was used to observe the pathological changes of renal tubules. TUNEL staining was applied to observe the apoptosis of rat kidney tissue. SIRT1 and FOXO3a mRNA expression levels in rat kidney tissue were measured by qPCR. Western blot assay was applied to detect the expression of SIRT1/FOXO3a pathway and autophagy-related proteins in rat kidney tissue. Results Compared with the control group, there was severe renal tissue pathological injury in the model group, and the 24 h urine microalbumin content, urine NAG, urine β2-MG, BUN, Scr, renal tubular injury score, renal MDA level and apoptosis rate were obviously increased. The renal SOD level, SIRT1/FOXO3a pathway and autophagy-related protein expression were obviously decreased (P<0.05). Compared with the model group, the pathological injury of kidney tissue was reduced in the NOB-L group and the NOB-H group, and the 24 h urine microalbumin content, urine NAG, urine β2-MG, BUN, Scr, renal tubular injury score, renal MDA level and apoptosis rate were obviously decreased. The renal SOD level, SIRT1/FOXO3a pathway and autophagy-related protein expression were obviously increased (P<0.05). Conclusion NOB may promote autophagy by activating the SIRT-1/FOXO3a pathway, thereby improving acute kidney injury in rats.

Key words: nobiletin, acute kidney injury, cisplatin, autophagy-related proteins, SIRT-1/FOXO3a

CLC Number:

R285.5

Figures/Tables 8

References 25

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基因名称	引物序列（5'→3'）	产物大小（bp）
SIRT1	上游：CAGTTCCAGCCATCTCTGTG	158
SIRT1	下游：GCAACCTGCTCCAAGGTATC	158
FOXO3a	上游：GCAAACCCTCTCGGACTCTC	116
FOXO3a	下游：TTCCCCACATTCAAACCAAC	116
GAPDH	上游：ACAGCAACAGGGTGGTGGAC	197
GAPDH	下游：TTTGAGGGTGCAGCG AACTT	197

基因名称	引物序列（5'→3'）	产物大小（bp）
SIRT1	上游：CAGTTCCAGCCATCTCTGTG	158
SIRT1	下游：GCAACCTGCTCCAAGGTATC	158
FOXO3a	上游：GCAAACCCTCTCGGACTCTC	116
FOXO3a	下游：TTCCCCACATTCAAACCAAC	116
GAPDH	上游：ACAGCAACAGGGTGGTGGAC	197
GAPDH	下游：TTTGAGGGTGCAGCG AACTT	197

组别	24 h尿微量白蛋白（mg）	尿NAG （U/L）	尿β2-MG （mg/L）
Control组	7.86±2.55	1.44±0.18	6.54±0.96
Model组	92.64±12.58^a	4.67±1.25^a	52.42±6.85^a
NOB-L组	68.25±9.68^b	3.37±0.59^b	34.66±4.20^b
NOB-H组	25.13±3.74^bc	1.95±0.34^bc	21.37±2.46^bc
NOB-H+EX527组	56.12±8.26^d	2.89±0.45^d	32.94±2.61^d
F	265.892^＊＊	60.064^＊＊	295.741^＊＊

组别	24 h尿微量白蛋白（mg）	尿NAG （U/L）	尿β2-MG （mg/L）
Control组	7.86±2.55	1.44±0.18	6.54±0.96
Model组	92.64±12.58^a	4.67±1.25^a	52.42±6.85^a
NOB-L组	68.25±9.68^b	3.37±0.59^b	34.66±4.20^b
NOB-H组	25.13±3.74^bc	1.95±0.34^bc	21.37±2.46^bc
NOB-H+EX527组	56.12±8.26^d	2.89±0.45^d	32.94±2.61^d
F	265.892^＊＊	60.064^＊＊	295.741^＊＊

组别	BUN（mmol/L）	Scr（μmol/L）
Control组	2.58±0.54	18.63±2.57
Model组	8.84±1.46^a	82.51±10.23^a
NOB-L组	6.72±1.34^b	60.16±7.44^b
NOB-H组	3.89±0.70^bc	42.56±6.30^bc
NOB-H+EX527组	5.24±1.25^d	56.27±8.82^d
F	109.769^＊＊	135.065^＊＊