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

Article information

J Korean Med. 2013;34(3):25-36

Publication date (electronic) : 2013 September 30

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

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 2013 March 15; Revised 2013 May 20; Accepted 2013 May 20.

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 Editionth ed. Seoul: Academy Press; 2000. p. 724.

3. J. CLAUDE bennett, FRED PLUM. Cecil textbook of medicine 20thth ed. 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.

Article information Continued

Fig. 1.

Effect of LPS on serum TNF-α level in SD rats.

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.

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.

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.