Effects of renal denervation on the vascular endothelial cell autophagy and NLRP3 activation in type 2 diabetic rats

doi:10.11958/20212359

Abstract

Abstract:

Objective To explore the effects of renal denervation (RDN) on the autophagy of vascular endothelial cells and NLRP3 inflammatory complexes in type 2 diabetic mellitus (T2DM) rats. Methods T2DM rat model was established by high-fat diet (HFD) combined with low-dose streptozotocin (STZ) intraperitoneal injection. Rats were randomly divided into four groups: the control group (CON, n=6), the diabetic group (T2DM, n=6), the bilateral sham surgery group (Sham, n=6) and the bilateral RDN group (RDN, n=6). Changes in all indicators were recorded at baseline, before and after RDN and 4 weeks after RDN, respectively. In vitro vascular ring tension test was used to evaluate vascular relaxation function. Western blot assay was used to detect the aorta endothelial cell autophagy of Beclin1, LC3 and p62, and the expression of NLRP3 protein and cysteine aspartate protease 1 (Caspase-1) and endothelial nitric oxide synthase (eNOS) protein. Results Compared with the control group, the endothelium diastolic function of the thoracic aorta was significantly decreased in the T2DM group, the expression of autophagy-related proteins Beclin1 and LC3 were decreased, the expression of p62 was increased. The expression levels of NLRP3, Caspase-1 and IL-1β protein were higher in the T2DM group than those of the control group (P<0.05), the level of eNOS protein was decreased (P<0.05). Compared with the T2DM group and the Sham group, the endothelium diastolic function was significantly improved in the RDN group, and the expression levels of Beclin1 and LC3 were increased, the expression of p62 was decreased. The protein expression levels of NLRP3, Caspase-1 and IL-1β were decreased, the level of eNOS increased (P<0.05). Conclusion The vascular endothelial dysfunction in T2DM rats may be associated with the decreased autophagy level and overexpression of NLRP3 inflammatory complex protein. RDN can enhance the autophagy in vascular endothelial cells, decrease the expression of NLRP3 inflammatory complex protein and improve vascular endothelial dysfunction.

Key words: diabetes mellitus, type 2, vasodilation, endothelial cells, NLR family, pyrin domain-containing 3 protein, autophagy, renal denervation

CLC Number:

R543

Figures/Tables 13

References 18

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基因名称	引物序列（5'→3'）	产物大小（bp）
NLRP3	上游：AGTCTTCGAGATTTCCTGGTAG	114
NLRP3	下游：CTTGTGCTCTGCTTGTGAGG	114
GAPDH	上游：CCTTCCGTGTTCCTACCC	130
GAPDH	下游：CCCAAGATGCCCTTCAGT	130

基因名称	引物序列（5'→3'）	产物大小（bp）
NLRP3	上游：AGTCTTCGAGATTTCCTGGTAG	114
NLRP3	下游：CTTGTGCTCTGCTTGTGAGG	114
GAPDH	上游：CCTTCCGTGTTCCTACCC	130
GAPDH	下游：CCCAAGATGCCCTTCAGT	130

组别	SBP（mmHg）	HR（次/min）	FPG（mmol/L）	FINS（IU/L)	NE（ng/L）	vWF（μg/L）	NO（μmol/L）
CON组	100.33±6.97	319.17±5.19	4.17±0.44	16.16±1.95	66.87±2.01	273.55±14.38	71.60±7.29
T2DM组	98.14±5.24	316.29±6.18	4.01±0.62	15.91±1.42	67.48±2.04	284.39±12.30	74.15±6.69
Sham组	102.28±4.86	314.43±4.43	4.31±0.37	17.21±1.79	66.50±2.27	281.41±14.61	72.96±5.79
RDN组	102.70±6.02	317.20±8.51	3.94±0.43	16.24±2.18	66.29±1.92	280.44±14.49	73.94±6.99
F	0.991	0.583	0.983	0.575	0.384	0.645	0.181

组别	SBP（mmHg）	HR（次/min）	FPG（mmol/L）	FINS（IU/L)	NE（ng/L）	vWF（μg/L）	NO（μmol/L）
CON组	100.33±6.97	319.17±5.19	4.17±0.44	16.16±1.95	66.87±2.01	273.55±14.38	71.60±7.29
T2DM组	98.14±5.24	316.29±6.18	4.01±0.62	15.91±1.42	67.48±2.04	284.39±12.30	74.15±6.69
Sham组	102.28±4.86	314.43±4.43	4.31±0.37	17.21±1.79	66.50±2.27	281.41±14.61	72.96±5.79
RDN组	102.70±6.02	317.20±8.51	3.94±0.43	16.24±2.18	66.29±1.92	280.44±14.49	73.94±6.99
F	0.991	0.583	0.983	0.575	0.384	0.645	0.181

组别	SBP（mmHg）	HR（次/min）	FPG（mmol/L）	FINS（IU/L)	NE（ng/L）	vWF（μg/L）	NO（μmol/L）
CON组	105.50±5.39	319.50±5.61	4.02±0.46	17.02±1.86	67.69±3.22	275.41±16.38	71.73±6.16
T2DM组	136.33±8.78^a	346.17±5.74^a	18.87±2.73^a	35.44±3.87^a	103.10±4.09^a	478.74±20.21^a	38.92±4.90^a
Sham组	135.50±5.99^a	348.83±7.31^a	19.45±2.60^a	33.61±3.33^a	105.01±3.73^a	491.75±18.51^a	37.56±4.18^a
RDN组	136.67±6.41^a	347.50±8.09^a	19.50±2.49^a	35.17±3.38^a	102.03±4.05^a	488.07±21.63^a	36.76±4.99^a
F	29.751^＊＊	25.813^＊＊	67.700^＊＊	46.338^＊＊	133.809^＊＊	179.718^＊＊	66.513^＊＊