Effect of Cordyceps sinensis on podocyte damage induced by high glucose by regulating the AMPK/mTOR pathway

doi:10.11958/20240816

Abstract

Abstract:

Objective To investigate effects of Cordyceps sinensis (CS) on high glucose (HG) induced podocyte injury by regulating the adenylate activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathway. Methods Mouse podocytes were cultured in vitro and divided into the normal glucose (NG) group, the HG group, the HG+CS group, the HG+CS+autophagy inhibitor (HG+CS+3MA) group and the HG+CS+AMPK inhibitor (HG+CS+Compound C) group. Podocyte viability was detected by CCK-8 method. Western blot assay was used to detect the expression levels of podocyte marker proteins podocin and nephrin, autophagy-related proteins Beclin-1 and P62, and pathway related proteins p-AMPK and p-mTOR. Results Compared with the NG group, the cell viability of podocytes decreased, the expression levels of podocin, nephrin，Beclin-1 and p-AMPK protein were decreased, and the expression levels of P62 and p-mTOR protein were increased in the HG group (P<0.05). Compared with the HG group, the cell viability of podocytes was increased, the expression levels of podocin, nephrin, Beclin-1 and p-AMPK protein were significantly increased (P<0.05), and the expression levels of P62 and p-mTOR protein were decreased in the HG+CS group (P<0.05). Compared with the HG+CS group, the cell viability decreased in the HG+CS+3MA group and the HG+CS+Compound C group (P<0.05). Compared with the HG+CS group, the HG+CS+3MA group and the HG+CS+Compound C group showed decreased expression levels of podocin, nephrin and Beclin-1 protein, and increased expression of P62 protein (P<0.05). Compared with the HG+CS group, the expression of p-AMPK protein decreased and the expression of p-mTOR protein increased in the HG+CS+Compound C group (P<0.05). Conclusion Cordyceps sinensis may play a protective role in diabetic nephropathy by up-regulating AMPK/mTOR signaling pathway to induce podocyte autophagy, alleviate high glucose induced podocyte injury and apoptosis.

Key words: cordyceps, diabetic nephropathies, autophagy, podocytes, AMPK/mTOR signaling pathway

CLC Number:

R285.5

Figures/Tables 7

Fig.1 Comparison of cell viability levels of podocytes between the five groups

Fig.2 The expression levels of podocin, nephrin, Beclin-1 and P62 in cells of each group under autophagy inhibitor intervention

Tab.1 Comparison of protein expression levels of podocin, nephrin, Beclin-1 and P62 under autophagy inhibitor intervention between the four groups of cells

组别	podocin	nephrin	Beclin-1	P62
NG组	1.00±0.00	1.00±0.01	1.00±0.00	1.00±0.00
HG组	0.27±0.03^a	0.32±0.05^a	0.33±0.04^a	4.62±0.57^a
HG+CS组	0.63±0.07^b	0.81±0.08^b	0.61±0.06^b	1.87±0.23^b
HG+CS+3MA组	0.20±0.02^c	0.20±0.05^c	0.25±0.05^c	3.61±0.23^c
F	454.211^＊	425.352^＊	322.695^＊	124.156^＊

Fig.3 The expression levels of p-AMPK and p-mTOR protein in each group affected by AMPK inhibitor intervention

Tab.2 Comparison of protein expression levels of p-AMPK and p-mTOR under AMPK inhibitor intervention between the four groups of cells

组别	p-AMPK/AMPK	p-mTOR/mTOR
NG组	1.00±0.00	1.00±0.01
HG组	0.35±0.03^a	1.40±0.05^a
HG+CS组	0.96±0.02^b	1.02±0.05^b
HG+CS+Compound C组	0.27±0.02^c	1.66±0.03^c
F	496.373^＊	305.433^＊

Fig.4 The expression levels of podocin, nephrin, Beclin-1 and P62 protein in each group affected by AMPK inhibitor intervention

Tab.3 Comparison of protein expression levels of podocin, nephrin, Beclin-1 and P62 under AMPK inhibitor intervention between the four groups of cells

组别	podocin	nephrin	Beclin-1	P62
NG组	1.00±0.00	1.00±0.00	1.00±0.00	1.00±0.00
HG组	0.20±0.03^a	0.43±0.05^a	0.23±0.04^a	4.40±0.67^a
HG+CS组	0.88±0.13^b	0.74±0.05^b	0.86±0.10^b	2.53±0.36^b
HG+CS+ Compound C组	0.30±0.05^c	0.31±0.04^c	0.40±0.07^c	4.26±0.47^c
F	151.175^＊	309.741^＊	160.780^＊	64.570^＊

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