Anti-inflammatory Effect of  Herbal-acupuncture at KI10 on LPS-induced Nephritis in Rats

Yang-seob Kim; Yun-Kyoung Yim; Kim, Yang-seob; Yim, Yun-Kyoung

doi:10.13048/jkm.13010

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

Original Article

Journal of Korean Medicine 2013; 34(3): 25-36.

Published online: September 30, 2013

DOI: https://doi.org/10.13048/jkm.13010

Anti-inflammatory Effect of Zeae Stigma Herbal-acupuncture at KI10 on LPS-induced Nephritis in Rats

Yang-seob Kim, Yun-Kyoung Yim

Dept. of Meridians & Acupoints, College of Oriental Medicine, Daejeon University

Correspondence to: 임윤경 (Yun-Kyoung Yim), 대전시 동구 용운동 대전대학교 한의학과 경락경혈학교실, Tel : +82-42-280-2610, Fax: +82-42-280-2641, E-mail: docwindy@dju.kr

Received March 15, 2013 Revised May 20, 2013 Accepted May 20, 2013

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.

Keywords: Zeae Stigma, herbal-acupuncture, KI10, LPS, nephritis

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

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

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

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

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

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

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

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

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

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

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

Scheme 1.

Manufacturing procedure of Zeae Stigma Herbal acupuncture Solution.

Scheme 2.

Experimental Procedure

References

1.. Du HK. Oriental Kidney System Internal Medicine. Seoul: Oriental Science Research Association;1993. p. 226p. 228p. 247p. 238–47. p. 326p. 334–6.

2.. Lee WJ. Medical Dictionary. The 2nd Edition. Seoul: Academy Press;2000. p. 724

3.. J. CLAUDE bennett, FRED PLUMCecil textbook of medicine. 20th. philadelphia: saunders;1996. p. 552p. 575–7.

4.. Frequent 22 Disease Classification Payroll Status. 2004–2011. Available at: URL:http://stat.kosis.kr/nsieu/view/tree.do?task=branchView&id=350_35001_6*MT_OTITLE&hOrg=350. Accessed October 2, 2012

5.. Acute Glomerulonephritis. 2005. Available at: URL: http://www.amc.seoul.kr/dept/bbs/vie-w.do?dtCode=D022&dtType=F&menuId=5897&id=62466. Accessed October 2, 2012

6.. Seo SJ. Diagnosis and Management of the Renal Diseases - Treatment of Glomerulonephritis. The Korean Journal of Medicine. 1962; 5:2. 81–5.

7.. Hong SK. Diagnosis and Management of the Renal Diseases - Theoretical Analysis of Plasma Clearance Concept. The Korean Journal of Medicine. 1962; 5:2. 75–8.

8.. Kim YI. Symposium: Management of the Idiopathic Nephrotic Syndrome in Adults. The Korean Journal of Medicine. 1978; 21:10. 811–9.

9.. Lee BY. Diagnosis and Management of the Renal Diseases: Nephrotic Syndrome. The Korean Journal of Medicine. 1962; 5:3. 141–4.

10.. Meridians & Acupoints Compilation Committee of Korean Oriental Medical Colleges. Details of Meridians & Acupoints; A Guidebook for College Students. Daejeon: Jongryeonamu Publishing Co;2012. p. 634–6.

11.. Committee of Korean Textbook Publisher. Herbology. Seoul: Yeong Lim Sa;2004. p. 353–4.

12.. Yoon HJ. The Glucoregulstory Effects of MAYDIS STIGMATA in Streptozotocin Induced Diabetic Rats. The Journal of Jeahan Oriental Medical Academy. 1995; 1:1. 178–87.

13.. Cho JH. Effect of Zeae Stigma on Renal injury induced by Gentamicin Sulfate in rats. J. of Herbology. 1994; 9:1. 143–57.

14.. Park BM, Hur B, Yim YK. Anti-inflammatory Effect of Plantaginis Semen Herbal-acupuncture at KI10 on LPS-induced nephritis in rats. Journal of Meridian & Acupoint. 2009; 26:2. 127–143.

15.. Cho E, Kang JH, Lee H. Anti-inflammatory Effect of Akebiae Lignum Parmacopuncture at KI10 on LPS-induced Acute Nephritis in Rats. The Journal of Korean Acupuncture & Moxibustion Society. 2012; 29:3. 41–53.

16.. Kim KM, Lee H, Kang HJ, Lee YH, Yim YK. Anti-inflammatory Effect of Dianthi Semen Herbal-acupuncture at KI10 on nephritis in rats. Journal of Meridian & Acupoint. 2009; 26:1. 61–77.

17.. Glauser MP, Zanetti G, Baumgartner JD, Cohen J. Septic shock Pathogenesis. Lancet. 1991; 338:732–6.

18.. Ohmori Y, Hamilton TA. Amacrophage LPS-inducible early gene encodes the murine homologue of IP-10. Biochem Biophys Res Commun. 1990; 168:1261–7.

19.. Tannenbaum CS, Koemer TJ, Jansen MM, Hamilton TA. Characteriation of Lipopolysaccharide-induced macrophage gene expression. J immunol. 1988; 140:3640–5.

20.. Jung HJ, Lim CG, Go GS, Ahn JH, Lee TW, Kim MJ, et al. Effects of Angiotensin - Converting Enzyme Inhibition (ACEi) in Serum HBsAg (+) Glomerulonephritis (GN)). The Korean Journal of Medicine. 1994; 46:6. 817–825.

21.. No JU. Therapeutic Guidelines of Rapidly progressive Glomerulonephritis, IgA Nephropathy, Membranoproliferative Glomerulonephritis. The Korean Journal of Nephrology. 2001; 20:suppl 3. 354–369.

22.. Lee KW. Blood pressure control in chronic kidney disease. 7. Available at: URL: http://www.medifonews.com/news/bbs.html?bcode=imsang. Accessed October 2, 2012

23.. Kang MS, Kim HR, Kim SK. Physiology. Seoul: Korea national Open University Press;2003. p. 72–76. p. 97–108.

24.. Kim JH, Yang KS. Effects of Polygonatum odoratum on Mercuric Chloride Induced Renal Failure Rats. Natutal Product Sciences. 2002; 33:3. 200–206.

25.. Nakagawa H, Komori N, Shibata F, Ikesue A, Konishi K, Fujioka M, et al. Identification of cytokineinduced neutrophil chemoattractants (CINC), rat GRO/CINC-2 alpha and CINC-2 beta, produced by granulation tissue in culture: purification, complete amino acid sequences and characterization. Biochem. J. 1994; 301:545–50.

26.. Taie S, Chujo K, Asaga T, Iwanaga Y, Ono J, Maekawa N. Urinary trypsin inhibitor reduces inflammatory response in kidney induced by lipopolysaccharide. Ueki J Biosci Bioeng. 2007; 104:4. 315–20.

27.. Lee KN, Kwon OH. Clinical Pathology File. Seoul: Medical Culture Press;2003. p. 95–8. p. 102–5. p. 1437