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J Acupunct Res > Volume 31(1); 2014 > Article
Han, Kim, Kim, and Yim: The Effect of Alismatis Rhizoma Herbal-acupuncture at KI10 on LPS-induced Nephritis in Rats
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Abstract

Objectives :

This study aimed to evaluate the effects of Alismatis Rhizoma Herbal-acupuncture(AR-HA) at KI10(Umgok) on nephritis induced by lipopolysaccharide(LPS) in rat.

Methods :

Rats were injected with LPS to induce nephritis. The rats in KI10-AR-HA group were treated with AR-HA, KI10-NP group with needle prick, KI10-saline group with normal saline injection respectively at KI10 three times for a week. Several experimental items were performed including measurements of the numbers of RBC, WBC, neutrophil in blood, the levels of creatinine, TNF-α, CINC-1 in serum, creatinine, total protein in urine, MPO in kidney.

Results :

Needle prick stimulation at KI10 significantly reduced WBC in blood of LPS-stimulated rats. Saline injection at KI10 significantly reduced WBC in blood, TNF-α in serum, total protein in urine of LPS-stimulated rats. AR-HA at KI10 significantly reduced WBC, neutrophil in blood, TNF-α, CINC-1 in serum, creatinine, total protein in urine of LPS-stimulated rats.

Conclusion :

AR-HA at KI10 has a therapeutic effect on nephritis in LPS-stimulated rat. There may be a synergism between KI10(Umgok) stimulation and AR-HA solution.

Fig. 1.
Effect of AR-HA at KI10 on RBC 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.
KI10-NP : LPS(2 mg/kg) challenge and a needle prick at KI10.
KI10-saline : LPS(2 mg/kg) challenge and saline(200 ul/rat) injection at KI10.
KI10-AR-HA : LPS(2 mg/kg) challenge and AR-HA(5 %, 200 ul/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. ‡: p<0.05 compared to NP group by Kruskal-wallis test.
acupunct-31-1-51-6f1.gif
Fig. 2.
Effect of AR-HA at KI10 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.
KI10-NP : LPS(2 mg/kg) challenge and a needle prick at KI10.
KI10-saline : LPS(2 mg/kg) challenge and saline(200 μl/rat) injection at KI10.
KI10-AR-HA : LPS(2 mg/kg) challenge and AR-HA(5 %, 200 μl/rat) at KI10.
*** : p<0.001 compared to normal group by Kruskal-wallis test.
††† : p<0.001, †† : p<0.01,
†: p<0.05 compared to LPS group by Kruskal-wallis test.
acupunct-31-1-51-6f2.gif
Fig. 3.
Effect of AR-HA at KI10 on neutrophil 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.
KI10-NP : LPS(2 mg/kg) challenge and a needle prick at KI10.
KI10-saline : LPS(2 mg/kg) challenge and saline(200 μl/rat) injection at KI10.
KI10-AR-HA : LPS(2 mg/kg) challenge and AR-HA(5 %, 200 μl/rat) at KI10.
††† : p<0.001 compared to LPS group by Kruskal-wallis test.
‡‡ : p<0.01 compared to N.P. group by Kruskal-wallis test.
## : p<0.01 compared to saline group by Kruskal-wallis test.
acupunct-31-1-51-6f3.gif
Fig. 4.
Effect of AR-HA at KI10 on serum TNF-α level in LPS-stimulated rats
Data were expressed as mean ± SD(n=5).
Normal : normal SD rat.
LPS : LPS(2 mg/kg) challenge.
KI10-NP : LPS(2 mg/kg) challenge and a needle prick at KI10.
KI10-saline : LPS(2 mg/kg) challenge and saline(200 μl/rat) injection at KI10.
KI10-AR-HA : LPS(2 mg/kg) challenge and AR-HA(5 %, 200 μl/rat) at KI10.
*** : p<0.001, ** : p<0.01 compared to normal group by Kruskal-wallis test.
†† : p<0.01, † : p<0.05 compared to LPS group by Kruskal-wallis test.
‡‡: p<0.01,
‡ : p<0.05 compared to NP group by Kruskal-wallis test.
acupunct-31-1-51-6f4.gif
Fig. 5.
Effect of AR-HA at KI10 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.
KI10-NP : LPS(2 mg/kg) challenge and a needle prick at KI10.
KI10-saline : LPS(2 mg/kg) challenge and saline(200 μl/rat) injection at KI10.
KI10-AR-HA : LPS(2 mg/kg) challenge and AR-HA(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-31-1-51-6f5.gif
Fig. 6.
Effect of AR-HA at KI10 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.
KI10-NP : LPS(2 mg/kg) challenge and a needle prick at KI10.
KI10-saline : LPS(2 mg/kg) challenge and saline(200 μl/rat) injection at KI10.
KI10-AR-HA : LPS(2 mg/kg) challenge and AR-HA(5 %, 200 μl/rat) at KI10.
†† : p<0.01 compared to LPS group by Kruskal-wallis test.
acupunct-31-1-51-6f6.gif
Fig. 7.
Effect of AR-HA at KI10 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.
KI10-NP : LPS(2 mg/kg) challenge and a needle prick at KI10.
KI10-saline : LPS(2 mg/kg) challenge and saline(200 μl/rat) injection at KI10.
KI10-AR-HA : LPS(2 mg/kg) challenge and AR-HA(5 %, 200 μl/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.
acupunct-31-1-51-6f7.gif
Fig. 8.
Effect of AR-HA at KI10 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.
KI10-NP : LPS(2 mg/kg) challenge and a needle prick at KI10.
KI10-saline : LPS(2 mg/kg) challenge and saline(200 μl/rat) injection at KI10.
KI10-AR-HA : LPS(2 mg/kg) challenge and AR-HA(5 %, 200 μl/rat) at KI10.
*** : p<0.001 compared to normal group by Kruskal-wallis test.
acupunct-31-1-51-6f8.gif
Scheme 1.
Manufacturing procedure of Alismatis Rhizoma herbal acupuncture solution
acupunct-31-1-51-6sc1.gif
Scheme 2.
Experimental procedure
acupunct-31-1-51-6sc2.gif
Table 1.
Reagents
Reagent Manufacturer Country
Phosphate buffer saline(PBS) USA
HBSS
Lipopolysaccharide(LPS) gum-sucrose absolute EtOH Sigma

Antibiotics Hyclone
FBS

BCA kit BD

TNF-alpha ELISA kit Oxford
Creatinine assay kit Bio Assay Systems
CINC-1 ELISA kit ALPCO
MPO ELISA kit Cellscience

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

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