Objective:

2

To evaluate the effects of combination of

Radix Astragali

(RA) and

Radix Salviae Miltiorrhizae

(RS) on kidney of spontaneously hypertensive rats (SHRs) and renal intrinsic cells.

Methods:

2

SHRs were intragastrically administrated with RA (5.09 g/kg) and RS (2.55 g/kg) either alone or with combination for 4 weeks; valsartan (13.35 mg/kg) was used as a positive control. Blood pressure and renal ultrasonography were monitored periodically. The biomarkers [microalbumin (mALB)

cystatin C

angiotensin Ⅱ (Ang Ⅱ)

interleukin-1 beta (IL-1β)

and β2-microglobulin (β2-Mg)

etc.] in serum and urine were measured by enzyme-linked immunosorbent assay (ELISA). The protein expressions [phosphorylated adenosine 5'-monophosphate-activated protein kinase-α1 (p-AMPKα1)

sestrin-β

calcium/calmodulin-dependent protein kinase kinase-β (CaMKK-β)

phosphoinositide 3-kinases (PI3K)

serine-threonine protein kinase 1 (AKT1)

and vascular endothelial growth factor receptor 2 (VEGFR2)]

in renal cortex were determined by Western blot.

In vitro

the hypertensive cellular model was established by applying 2×10

-6

mol/L Ang Ⅱ. The primary human podocytes

human glomerular endothelial cells (HRGECs)

and human proximal tubular epithelial cells (HK-2s) were pre-incubated with sulfotanshinone sodium (Tan

10 μg/mL) and/or calycosin-7-O-β-D-glucoside (Cal

5 μg/mL). The cellular viability and apoptosis were assayed by 3-(4

5-Dimethylthiazol-2-yl)-2

5-diphenyltetrazolium bromide (MTT) and Annexin V/PI staining

respectively. The level of endothelial nitric oxide synthase (eNOS) in culture supernatant was determined by ELISA.

Results:

2

RA+RS significantly decreased the diastolic blood pressure

renal vascular resistance index

and parenchymal thickness

increased 24 h urinary volume as well as lowered the levels of urine mALB and serum cystatin C

IL-1β and β2-Mg of SHRs (

P

<

0.05 vs. SHRs). The decreased protein levels of p-AMPKα1

sestrinβ and CaMKK-β and the increased protein levels of PI3K

AKT1 and VEGFR2 in renal cortex of SHRs were normalized after RA+RS treatment (

P

<

0.05).

In vitro

Tan and Cal attenuated the Ang Ⅱ-induced abnormal proliferation and increased the apoptosis of HRGECs and HK-2s and improved the level of eNOS in culture supernatant. Whereas

neither of them showed powerful effect on podocyte.

Conclusion:

2

The combination of RA and RS had potential effects on alleviating the renal damages of SHRs and the renoprotection was independent of blood pressure level.