Tetrandrine may treat membranous glomerulopathy via P13K/Akt signaling pathway regulation: therapeutic mechanism validation using Heymann nephritis rat model

Figures & data

Table 1. The main targets of tetrandrine in the treatment of nephrotic syndrome (NS)

ID	Name	ID	Name	ID	Name	ID	Name
28	ABO	1991	ELANE	3718	JAK3	142	PARP1
207	AKT1	2099	ESR1	8850	KAT2B	5105	PCK1
213	ALB	2224	FDPS	3791	KDR	8654	PDE5A
308	ANXA5	2539	G6PD	3815	KIT	5294	PIK3CG
353	APRT	2739	GLO1	3932	LCK	5328	PLAU
23,621	BACE1	2811	GP1BA	3934	LCN2	5566	PRKACA
598	BCL2L1	2885	GRB2	4048	LTA4H	5660	PSAP
635	BHMT	2936	GSR	4069	LYZ	5879	RAC1
673	BRAF	2944	GSTM1	11,253	MAN1B1	5916	RARG
695	BTK	2950	GSTP1	1432	MAPK14	5950	RBP4
834	CASP1	3074	HEXB	5599	MAPK8	5972	REN
836	CASP3	3156	HMGCR	4193	MDM2	6256	RXRA
890	CCNA2	27,306	HPGDS	4233	MET	6401	SELE
1017	CDK2	3251	HPRT1	4282	MIF	6714	SRC
1022	CDK7	3265	HRAS	4311	MME	6772	STAT1
629	CFB	3290	HSD11B1	4321	MMP12	7068	THRB
1118	CHIT1	3320	HSP90AA1	4314	MMP3	7276	TTR
1384	CRAT	3479	IGF1	4318	MMP9	7421	VDR
1495	CTNNA1	3558	IL2	4846	NOS3	331	XIAP
1508	CTSB	3643	INSR	2908	NR3C1	7535	ZAP70
1513	CTSK	3683	ITGAL	5009	OTC
1956	EGFR	3717	JAK2	23,569	PADI4

Figure 1. Venn diagram of targets for tetrandrine treating MGN

Figure 2. The nerwork of MGN-related targets

Table 2. The top 15 KEGG pathways of tetrandrine related target genes

ID	Description	p.adjust	Count
hsa05215	Prostate cancer	9.61E-10	13
hsa01522	Endocrine resistance	9.61E-10	13
hsa05205	Proteoglycans in cancer	9.61E-10	17
hsa04151	PI3K-Akt signaling pathway	1.24E-09	21
hsa01521	EGFR tyrosine kinase inhibitor resistance	1.29E-08	11
hsa05161	Hepatitis B	2.22E-08	14
hsa04068	FoxO signaling pathway	1.87E-07	12
hsa05418	Fluid shear stress and atherosclerosis	3.23E-07	12
hsa04917	Prolactin signaling pathway	6.75E-07	9
hsa05166	HTLV-I infection	6.75E-07	14
hsa04933	AGE-RAGE signaling pathway in diabetic complications	1.06E-06	10
hsa04014	Ras signaling pathway	1.17E-06	14
hsa04659	Th17 cell differentiation	1.72E-06	10
hsa04015	Rap1 signaling pathway	2.33E-06	13
hsa04010	MAPK signaling pathway	2.73E-06	15

Figure 3. GO and KEGG enrichment analysis. (a) The top 15 significantly enriched terms in biological process (BP); (b) The top 15 significantly enriched terms in cellular component (CC); (c) The top 15 significantly enriched terms in molecular function (MF); (d) The top 15 significantly enriched terms in KEGG pathway. Gene ratio = count/set size

Figure 4. The PPI network of 10 hub genes. IGF1: Insulin-like growth factor 1; AKT1: AKT Serine/Threonine Kinase 1; CASP3: Caspase 3; HRAS: HRas Proto-Oncogene, GTPase; ESR1: Estrogen Receptor 1; MAPK8: Mitogen-Activated Protein Kinase 8; MMP9: Matrix Metallopeptidase 9; SRC: SRC Proto-Oncogene, Non-Receptor tyrosine kinase; EGFR: Epidermal Growth Factor Receptor; ALB: Albumin

Figure 5. The top 15 GO terms of hub genes

Figure 6. The top 15 pathways of hub genes

Figure 7. Compound action disease targets were associated with the top 15 KEGG pathways and the top 15 GO terms

Figure 8. (a) The chemical structure of tetrandrine. (b) The 24 UP contents were detected after 7, 14, 21 and 28 days of tetrandrine treat in control, HN model and tetrandrine groups. (c) The representative images of the histology staining on the kidney tissue. (d) The representative images of TEM on the kidney tissue. **, P < 0.01 vs. control group; ##, P < 0.01 vs. model group

Figure 9. After 7, 14, 21 and 28 days of tetrandrine treat, the serum content of ALB (a), SCr (b), ALT (c) and AST (d) level in control, model and tetrandrine groups. *, P < 0.05 and **, P < 0.01 vs. control group; #, P < 0.05 and ##, P < 0.01 vs. model group

Table 3. The anti-oxidative effect of tetrandrine in Heymann nephritis (HN) rats

		Control	Model	Tetrandrine
Kidney	SOD (U/mL)	145.67 ± 9.03	78.55 ± 9.91**	113.63 ± 5.52 ^##
	MDA (nmol/mL)	8.16 ± 0.34	12.46 ± 0.58**	9.73 ± 0.42 ^##
	GSH-px (μmol/L)	5521.40 ± 505.74	2861.92 ± 557.51**	4408.34 ± 378.48 ^#
Serum	SOD (U/mL)	356.94 ± 30.35	216.42 ± 24.96**	281.79 ± 17.10 ^#
	MDA (nmol/mL)	8.20 ± 0.49	16.36 ± 0.87**	11.49 ± 0.76 ^##
	GSH-px (μmol/L)	543.55 ± 21.82	257.49 ± 28.20**	418.39 ± 35.31^##

The levels of MDA and the activities of SOD and GSH-Px in serum and kidney of HN and control rats were detected after 4-week administration of tetrandrine. ** P < 0.01 vs. control rats; # P < 0.05 and ## P < 0.01 vs. HN rats.

Figure 10. (a) Nephrin and podocin expression of kidney tissues were detected by immunofluorescence staining. (b) The kidney tissues cell apoptosis was analyzed via TUNEL

Figure 11. (a) The expression levels of IL-6, TNFα and MCP-1 in the kidney tissues were examined by qRT-PCR in control, model and tetrandrine groups rats. (b) The expression of p-PI3K, PI3K, Akt, p-Akt, p-BAD, Bcl-2 were examined by western blot in control, model and tetrandrine groups rats. **, P < 0.01 vs. control group; ##, P < 0.01 vs. model group

Data availability

The data used to support the findings of this study are available from the corresponding author upon request.