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J Acupunct Res > Volume 32(3); 2015 > Article
Jang, Kim, and Yim: The Effect of Pyrrosiae Herba Herbal-acupuncture at KI10 on Lipopolysaccharide Induced Nephritis in Rats
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Abstract

Objectives:

The purpose of this study is to investigate the effects of Pyrrosiae Herba herbal-acupuncture(PH-HA) at KI10(Umgok) on nephritis induced by lipopolysaccharide(LPS) in rats.

Methods:

Rats were assigned to four groups: normal, LPS, saline and PH-HA. Rats in the saline and PH-HA groups were treated with saline injection and PH-HA respectively at KI10, three times over the period of one week. All animals, except those in the normal group, were injected intra-peritoneally with LPS to induce nephritis. WBC, in blood, tumor necrosis factor alpha(TNF-α), cytokine-induced neutrophil chemoattractant 1(CINC-1), blood urea nitrogen(BUN), creatinine in serum, urinal volume, total protein creatinine in urine, and renal myeloperoxidase (MPO) were analyzed.

Results:

  1. PH-HA group showed significantly reduced levels of serum BUN, serum creatinine, TNF-α, and CINC-1 compared to the LPS group. Furthermore, a significant increase in urine output and more significant decreases in total protein in urine and MPO in renal tissue were observed in the PH-HA group when compared to the LPS group.

  2. The PH-HA group showed significantly reduced levels of serum creatinine and renal MPO, and a more significant increase in urine output compared to the saline group.

Conclusions:

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

Fig. 1
Manufacturing procedure of Pyrrosiae Herba herbal acupuncture solution
acupunct-32-3-15f1.gif
Fig. 2
Experimental procedure
acupunct-32-3-15f2.gif
Fig. 3
Effect of PH-HA on WBC count in blood of LPS-stimulated rats
Data were expressed as mean ± SD(n=5).
Normal: normal SD rat.
LPS: LPS(2 mg/kg) challenge.
Saline: LPS(2 mg/kg) challenge and saline(200 μl/rat) injection at KI10.
PH-HA: LPS(2 mg/kg) challenge and PH-HA(2.5 %, 200 μl/rat) at KI10.
**: p>0.01 compared to normal group by Kruskal-Wallis test.
acupunct-32-3-15f3.gif
Fig. 4
Effects of PH-HA on serum BUN level in LPS-stimulated rats
Data were expressed as mean ± SD(n=5).
Normal: normal SD rat
LPS: LPS(2 mg/kg) challenge.
Saline: LPS(2 mg/kg) challenge and saline(200 μl/rat) injection at KI10.
PH-HA: LPS(2 mg/kg) challenge and PH-HA(2.5 %, 200 μl/rat) at KI10.
**: p<0.01 compared to normal group by Kruskal-Wallis test.
†††: p<0.001 ††: p<0.01 compared to LPS group by Kruskal-Wallis test.
acupunct-32-3-15f4.gif
Fig. 5
Effects of PH-HA on serum creatinine level in LPS-stimulated rats
Data were expressed as mean ± SD(n=5).
Normal: normal SD rat.
LPS: LPS(2 mg/kg) challenge.
Saline: LPS(2 mg/kg) challenge and saline(200 μl/rat) injection at KI10.
PH-HA: LPS(2 mg/kg) challenge and PH-HA(2.5 %, 200 μl/rat) at KI10.
***: p<0.001 compared to normal group ANOVA test.
†††: p<0.001, compared to LPS group by Kruskal-Wallis test.
###: p<0.001 compared to saline group by Kruskal-Wallis test.
acupunct-32-3-15f5.gif
Fig. 6
Effects of PH-HA on serum TNF-alpha level in LPS-stimulated rats
Data were expressed as mean ± SD(n=5).
Normal: normal SD rat.
LPS: LPS(2 mg/kg) challenge.
Saline: LPS(2 mg/kg) challenge and saline(200 μl/rat) injcetion at KI10.
PH-HA: LPS(2 mg/kg) challenge and PH-HA(2.5 %, 200 μl/rat) at KI10.
***: p<0.001 **: p<0.01 *: p<0.05 compared to normal group by Kruskal-Wallis test.
††: p<0.01 compared to LPS group by Kruskal-wallis test.
acupunct-32-3-15f6.gif
Fig. 7
Effects of PH-HA on serum CINC-1 level in LPS-stimulated rats
Data were expressed as mean ± SD(n=5).
Normal: normal SD rat.
LPS: LPS(2 mg/kg) challenge.
Saline: LPS(2 mg/kg) challenge and saline(200 μl/rat) injcetion at KI10.
PH-HA: LPS(2 mg/kg) challenge and PH-HA(2.5 %, 200 μl/rat) at KI10.
**: p<0.01 compared to normal group by Kruskal-Wallis test.
††: p<0.01 compared to LPS group by Kruskal-wallis test.
acupunct-32-3-15f7.gif
Fig. 8
Effect of PH-HA on urinary volume in LPS-stimulated rats
Data were expressed as mean ± SD(n=3).
Normal: normal SD rat.
LPS: LPS(2 mg/kg) challenge.
Saline: LPS(2 mg/kg) challenge and saline(200 μl/rat) injection at KI10.
PH-HA: LPS(2 mg/kg) challenge and PH-HA(2.5 %, 200 μl/rat) at KI10.
**: p<0.01 *: p<0.05 compared to normal group by Kruskal-Wallis test
††: p<0.01 compared to LPS group by Kruskal-Wallis test.
##: p<0.01 compared to saline group by Kruskal-Wallis test.
acupunct-32-3-15f8.gif
Fig. 9
Effects of PH-HA on urinary creatinine level in LPS-stimulated rats
Data were expressed as mean ± SD(n=3).
Normal: normal SD rat.
LPS: LPS(2 mg/kg) challenge.
Saline: LPS(2 mg/kg) challenge and saline(200 μl/rat) injection at KI10.
PH-HA: LPS(2 mg/kg) challenge and PH-HA(2.5 %, 200 μl/rat) at KI10.
*: p<0.05 compared to normal group by Kruskal-Wallis test.
acupunct-32-3-15f9.gif
Fig. 10
Effects of PH-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(2 mg/kg) challenge.
Saline: LPS(2 mg/kg) challenge and saline(200 μl/rat) injection at KI10.
PH-HA: LPS(2 mg/kg) challenge and PH-HA(2.5 %, 200 μl/rat) at KI10.
**: p<0.01 compared to normal group by Kruskal-Wallis test.
†: p<0.05 compared to LPS group by Kruskal-Wallis test.
acupunct-32-3-15f10.gif
Fig. 11
Effects of PH-HA on renal MPO level in LPS-stimulated rats
Data were expressed as mean ± SD(n=5).
Normal: normal SD rat.
LPS: LPS(2 mg/kg) challenge.
Saline: LPS(2 mg/kg) challenge and saline(200 μl/rat) injection at KI10.
PH-HA: LPS(2 mg/kg) challenge and PH-HA(2.5 %, 200 μl/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.
#: p<0.05 compared to saline group by Kruskal-Wallis test.
acupunct-32-3-15f11.gif
Table 1
Reagents
Reagent Manufacturer Country
Phosphate buffer saline(PBS) Sigma USA
Hanks’ balanced salt solution(HBBS)
Lipopolysaccharide(LPS) gum-sucrose absolute ethanol

Antibiotics Hyclone
Fetal bovine serum(FBS)

Bicinchoninic Acid(BCA) kit Becton, Dickinson and Company(BD)

Tumor necrosis factor alpha(TNF-α)
 Enzyme-linked immunosorbent assay(ELISA) kit Oxford
Bio Assay Systems
ALPCO
Cellscience
Creatinine assay kit
Cytokine-induced neutrophil chemoattractant 1(CINC-1) ELISA Kit
Myeloperoxidase(MPO) ELISA Kit

Ethyl ether
Table 2
Instruments
Instruments Manufacturer Country
Rotary evaporator Eyela Japan
ELISA reader TARAN Canada
Sonicate Fisher USA
Metabolic cage B&P Korea
Cytological centrifuge Hanil
Micro centrifuge
VI.
VI.

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