Effects of Polyporus Herbal-acupuncture at KI10 on LPS-induced nephritis in rats

Article information

J Korean Med. 2015;36(3):73-83
1Department of Meridian & Acupoint, College of korean Medicine, Daejeon University
Correspondence to: 임윤경(Yun-Kyoung Yim), 대전시 동구 용운동 96-3 대전대학교 한의과대학 경락경혈학교실 12407호, Tel: +82-42-280-2610, Fax: +82-42-280-2647, E-mail: docwindy@dju.kr
Received 2015 June 08; Revised 2015 September 25; Accepted 2015 September 25.

Abstract

Objectives:

The purpose of this study was to evaluate the effects of Polyporus Herbal-acupuncture(PO-HA) at KI10 (Umgok) on nephritis induced by lipopolysaccharide(LPS) in rats.

Methods:

Rats were allocated into normal, control, and 2 experimental groups. The rats in the control group were intra-peritoneally injected with LPS for nephritis induction. The rats in the groups of experiment 1 and experiment 2 were treated with Saline injection, and PO-HA, respectively at KI10 three times for a week and then intra-peritoneally injected with LPS. To evaluate the effects of PO-HA at KI10, WBC count in blood, creatine, tumor necrosis factor-alpha (TNF-α), cytokine-induced neutrophil chemoattractant 1(CINC-1) in serum, urinary volume, creatinine, total protein in urine, myeloperoxidase(MPO) in kidney were measured.

Results:

PO-HA at KI10 significantly suppressed the increase of WBC in blood, TNF-α, CINC-1 in serum, MPO in kidney of LPS-stimulated rats. PO-HA at KI10 significantly suppressed the increase creatinine, total protein in urine of LPS-stimulated rats.

Conclusions:

According to these results, it is postulated that PO-HA at KI10 has an anti-inflammatory and renal-protective effects on LPS-induced nephritis in rats. Therefore, it is suggested that PO-HA at KI10 may be an useful therapeutics for nephritis in clinical field after further researches.

Fig. 1.

Cytotoxicity of PO-HAS on rat liver cells.

Liver cells from normal SD rat were cultured in RPMI 1640 with 10% FBS medium for 72h with or without various concentrations of PO-HAS (Polyporus Herbal-acupuncture solution). The cell viability were measured by MTT assay.

Values represent the means ± SD of 3 independent experiments. Control : culture medium without PO-HAS

10%, 2.5%, 1% PO-HAS : culture medium with 10%, 2.5%, 1% PO-HAS respectively

Fig. 2.

Effect of PO-HA on WBC count in blood of LPS-stimulated rats.

Data were expressed as mean ± SD (n=5).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10

PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

*** : p<0.001 compared to normal group by Kruskal-wallis test

††† : p<0.01 † : p<0.05 compared to LPS group by Kruskal -wallis test

# : p<0.05 compared to Saline group by Kruskal-wallis test

Fig. 3.

Effects of PO-HA on serum creatinine level in LPS-stimulated rats.

Data were expressed as mean ± SD (n=5).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10

PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

*** : p<0.001 compared to normal group by Kruskal-wallis test

## : p<0.01 compared to Saline group by Kruskal-wallis test

Fig. 4.

Effects of PO-HA on serum TNF-alpha level in LPS-stimulated rats.

Data were expressed as mean ± SD (n=5).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10

PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

*** : p<0.001 * : p<0.05 compared to normal group by Kruskal-wallis test

†† : p<0.01 compared to LPS group by Kruskal-wallis test

Fig. 5.

Effects of PO-HA on serum CINC-1 level in LPS-stimulated rats.

Data were expressed as mean ± SD (n=5).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10 PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

*** : p<0.001 ** : p<0.01 compared to normal group by Kruskal -wallis test

†† : p<0.01 compared to LPS group by ANOVA test

Fig. 6.

Effect of PO-HA on urinary volume in LPS-stimulated rats.

Data were expressed as mean ± SD (n=3).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10

PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

** : p<0.01 * : p<0.05 compared to normal group by Kruskal-wallis test

Fig. 7.

Effects of PO-HA on urinary creatinine level in LPS-stimulated rats.

Data were expressed as mean ± SD (n=3).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10

PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

* : p<0.05 compared to normal group by Kruskal-wallis test

††† : p<0.001 compared to LPS group by Kruskal-wallis test

## : p<0.01 compared to Saline group by Kruskal-wallis test

Fig. 8.

Effects of PO-HA on total protein level in urine of LPS-stimulated rats.

Data were expressed as mean ± SD (n=3).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10

PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

* : p<0.05 compared to normal group by Kruskal-wallis test

† : p<0.05 compared to LPS group by Kruskal-wallis test

Fig. 9.

Effects of PO-HA on renal MPO level in LPS-stimulated rats.

Data were expressed as mean ± SD (n=5).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10

PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

*** : p<0.001 compared to normal group by Kruskal-wallis test

†† : p<0.01 compared to LPS group by Kruskal-wallis test

Scheme 1.

Manufacturing procedure of Polyporus Herbal-acupuncture Solution.

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Article information Continued

Fig. 1.

Cytotoxicity of PO-HAS on rat liver cells.

Liver cells from normal SD rat were cultured in RPMI 1640 with 10% FBS medium for 72h with or without various concentrations of PO-HAS (Polyporus Herbal-acupuncture solution). The cell viability were measured by MTT assay.

Values represent the means ± SD of 3 independent experiments. Control : culture medium without PO-HAS

10%, 2.5%, 1% PO-HAS : culture medium with 10%, 2.5%, 1% PO-HAS respectively

Fig. 2.

Effect of PO-HA on WBC count in blood of LPS-stimulated rats.

Data were expressed as mean ± SD (n=5).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10

PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

*** : p<0.001 compared to normal group by Kruskal-wallis test

††† : p<0.01 † : p<0.05 compared to LPS group by Kruskal -wallis test

# : p<0.05 compared to Saline group by Kruskal-wallis test

Fig. 3.

Effects of PO-HA on serum creatinine level in LPS-stimulated rats.

Data were expressed as mean ± SD (n=5).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10

PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

*** : p<0.001 compared to normal group by Kruskal-wallis test

## : p<0.01 compared to Saline group by Kruskal-wallis test

Fig. 4.

Effects of PO-HA on serum TNF-alpha level in LPS-stimulated rats.

Data were expressed as mean ± SD (n=5).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10

PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

*** : p<0.001 * : p<0.05 compared to normal group by Kruskal-wallis test

†† : p<0.01 compared to LPS group by Kruskal-wallis test

Fig. 5.

Effects of PO-HA on serum CINC-1 level in LPS-stimulated rats.

Data were expressed as mean ± SD (n=5).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10 PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

*** : p<0.001 ** : p<0.01 compared to normal group by Kruskal -wallis test

†† : p<0.01 compared to LPS group by ANOVA test

Fig. 6.

Effect of PO-HA on urinary volume in LPS-stimulated rats.

Data were expressed as mean ± SD (n=3).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10

PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

** : p<0.01 * : p<0.05 compared to normal group by Kruskal-wallis test

Fig. 7.

Effects of PO-HA on urinary creatinine level in LPS-stimulated rats.

Data were expressed as mean ± SD (n=3).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10

PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

* : p<0.05 compared to normal group by Kruskal-wallis test

††† : p<0.001 compared to LPS group by Kruskal-wallis test

## : p<0.01 compared to Saline group by Kruskal-wallis test

Fig. 8.

Effects of PO-HA on total protein level in urine of LPS-stimulated rats.

Data were expressed as mean ± SD (n=3).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10

PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

* : p<0.05 compared to normal group by Kruskal-wallis test

† : p<0.05 compared to LPS group by Kruskal-wallis test

Fig. 9.

Effects of PO-HA on renal MPO level in LPS-stimulated rats.

Data were expressed as mean ± SD (n=5).

Normal : normal SD rat

LPS : LPS (2mg/kg) challenge

Saline : LPS (2mg/kg) challenge and Saline(200μℓ/rat) injection at KI10

PO-HA : LPS (2mg/kg) challenge and PO-HA(10%, 200μℓ/rat) at KI10

*** : p<0.001 compared to normal group by Kruskal-wallis test

†† : p<0.01 compared to LPS group by Kruskal-wallis test

Scheme 1.

Manufacturing procedure of Polyporus Herbal-acupuncture Solution.