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JKM > Volume 34(3); 2013 > Article
Kim and Yim: Anti-inflammatory Effect of Zeae Stigma Herbal-acupuncture at KI10 on LPS-induced Nephritis in Rats

Abstract

Objectives:

The purpose of this study was to investigate the effects of Zeae Stigma herbal-acupuncture (ZS-HA) at KI10 (Umgok) on nephritis induced by lipopolysaccharide (LPS) in rats.

Methods:

Rats were assigned to five groups, normal, LPS, NP, saline and ZS-HA. Rats in NP, saline, and ZS-HA groups were treated with needle prick, saline injection, and ZS-HA respectively at KI10, three times a week. All animals except the normal group were injected intra-peritoneally with LPS to induce nephritis. RBC and WBC, neutrophils in blood, TNF-alpha, CINC-1, creatinine in serum, urinal volume, total protein and creatinine in urine, and renal MPO were analyzed.

Results:

Needle prick at KI10 significantly reduced WBC in blood and CINC-1 in serum of LPS-stimulated rats. Saline injection at KI10 significantly reduced TNF-α in serum and total protein in urine of LPS-stimulated rats. ZS-HA at KI10 significantly increased RBC in blood, and significantly reduced neutrophils in blood, TNF-α in serum, and total protein in urine of LPS-stimulated rats.

Conclusions:

According to these results, it is postulated that ZS-HA at KI10 has anti-inflammatory and renal-protective effects on LPS-induced nephritis in rats, and both acupoint KI10 and herb Zeae Stigma made contributions to this effect. Further studies on the interaction between acupoint KI10 and herb Zeae Stigma may be needed.

Fig. 1.
Effect of LPS on serum TNF-α level in SD rats.
The male SD rats were injected intra-peritoneally with LPS(lipopolysaccharide; 2mg/kg). Blood samples were taken from rat heart at 1 hr or 3 hr after the LPS injection, and serum TNF-α level was analysed by ELISA.
Data were expressed as mean ± SD (n=3).
No treatment : normal SD rat.
LPS 2 mg/kg: SD rat with LPS (2mg/kg) challenge
* : p<0.05, compared to no-treatment group by t-test
jkm-34-3-25-3f1.tif
Fig. 2.
Effect of LPS on renal TNF-α level in SD rats.
The male SD rats were injected intra-peritoneally with LPS (2mg/kg). Rat kidney was removed at 1 hr or 3 hr after the LPS injection, and renal TNF-α level was analysed by ELISA. Data were expressed as mean ± SD (n=3).
No treatment : normal SD rat without LPS challenge
LPS 2 mg/kg: SD rat with LPS (2mg/kg) challenge
** : p<0.001, compared to no-treatment group by t-test
jkm-34-3-25-3f2.tif
Fig. 3.
Effects of ZS-HA on RBC count in blood of LPS-stimulated rats.
The male SD rats were treated as described in the materials and method and injected intra-peritoneally with LPS (2mg/kg). Blood samples were taken from rat hearts 1 hr after the LPS injection and RBC count was analysed.
Data were expressed as mean ± SD (n=5).
Normal : normal SD rat
LPS : LPS (2mg/kg) challenge
N.P.: LPS (2mg/kg) challenge and a needle prick at KI10
Saline : LPS (2mg/kg) challenge and saline(200μl/rat) injection at KI10
ZS-HA : LPS (2mg/kg) challenge and ZS-HA(10%, 200μl/rat) at KI10
* : p<0.05 compared to normal group by ANOVA test
† : p<0.05 compared to LPS group by ANOVA test
‡ : p<0.05 compared to N.P. group by ANOVA test
jkm-34-3-25-3f3.tif
Fig. 4.
Effect of ZS-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
N.P.: LPS (2mg/kg) challenge and a needle prick at KI10
Saline : LPS (2mg/kg) challenge and saline(200μl/rat) injection at KI10
ZS-HA : LPS (2mg/kg) challenge and ZS-HA(10%, 200μl/rat) at KI10
** : p<0.01 compared to normal group by ANOVA test
†† : p<0.01 compared to LPS group by ANOVA test
jkm-34-3-25-3f4.tif
Fig. 5.
Effect of ZS-HA on neutrophil count in blood of LPS-stimulated rats.
Data were expressed as mean ± SD (n=5).
Normal : normal SD rat
LPS : LPS (2mg/kg) challenge
N.P.: LPS (2mg/kg) challenge and a needle prick at KI10
Saline : LPS (2mg/kg) challenge and saline(200μl/rat) injection at KI10
ZS-HA : LPS (2mg/kg) challenge and ZS-HA(10%, 200μl/rat) at KI10
** : p<0.01 * : p<0.05 compared to normal group by ANOVA test
††† : p<0.001 compared to LPS group by ANOVA test
‡‡‡ : p<0.001 compared to N.P. group by ANOVA test
### : p<0.001 compared to saline group by ANOVA test
jkm-34-3-25-3f5.tif
Fig. 6.
Effects of ZS-HA on serum creatine level in LPS-stimulated rats.
Data were expressed as mean ± SD (n=5).
Normal : normal SD rat
LPS : LPS (2mg/kg) challenge
N.P.: LPS (2mg/kg) challenge and a needle prick at KI10
Saline : LPS (2mg/kg) challenge and saline(200μl/rat) injection at KI10
ZS-HA : LPS (2mg/kg) challenge and ZS-HA(10%, 200μl/rat) at KI10
** : p<0.01 * : p<0.05 compared to normal group ANOVA test
jkm-34-3-25-3f6.tif
Fig. 7.
Effects of ZS-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
N.P.: LPS (2mg/kg) challenge and a needle prick at KI10
Saline : LPS (2mg/kg) challenge and saline(200μl/rat) injection at KI10
ZS-HA : LPS (2mg/kg) challenge and ZS-HA(10%, 200μl/rat) at KI10
*** : p<0.001 * : p<0.05 compared to normal group by ANOVA test
† : p<0.05 compared to LPS group by ANOVA test
‡ : p<0.05 compared to N.P. group by ANOVA test
jkm-34-3-25-3f7.tif
Fig. 8.
Effects of ZS-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
N.P.: LPS (2mg/kg) challenge and a needle prick at KI10
Saline : LPS (2mg/kg) challenge and saline(200/μl/rat) injection at KI10
ZS-HA : LPS (2mg/kg) challenge and ZS-HA(10%, 200μl/rat) at KI10
** : p<0.01 compared to normal group by ANOVA test
† : p<0.05 compared to LPS group by ANOVA test
jkm-34-3-25-3f8.tif
Fig. 9.
Effects of ZS-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
N.P.: LPS (2mg/kg) challenge and a needle prick at KI10
Saline : LPS (2mg/kg) challenge and saline(200μl/rat) injection at KI10
ZS-HA : LPS (2mg/kg) challenge and ZS-HA(10%, 200μl/rat) at KI10
jkm-34-3-25-3f9.tif
Fig. 10.
Effects of ZS-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
N.P.: LPS (2mg/kg) challenge and a needle prick at KI10
Saline : LPS (2mg/kg) challenge and saline(200μl/rat) injection at KI10
ZS-HA : LPS (2mg/kg) challenge and ZS-HA(10%, 200μl/rat) at KI10
* : p<0.05 compared to normal group by ANOVA test
†† : p<0.01 † : p<0.05 compared to LPS group by ANOVA test
jkm-34-3-25-3f10.tif
Fig. 11.
Effects of ZS-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
N.P.: LPS (2mg/kg) challenge and a needle prick at KI10
Saline : LPS (2mg/kg) challenge and saline(200μl/rat) injection at KI10
ZS-HA : LPS (2mg/kg) challenge and ZS-HA(10%, 200μl/rat) at KI10
*** : p<0.001 compared to normal group by ANOVA test
jkm-34-3-25-3f11.tif
Scheme 1.
Manufacturing procedure of Zeae Stigma Herbal acupuncture Solution.
jkm-34-3-25-3f12.tif
Scheme 2.
Experimental Procedure
jkm-34-3-25-3f13.tif

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