The American Journal of Chinese Medicine, Vol. 35, No.

2, 241–250
© 2007 World Scientific Publishing Company
Institute for Advanced Research in Asian Science and Medicine

The Effect of Oculo-Acupuncture on Recovery

from Ethylene Glycol-Induced Acute Renal
Injury in Dogs
Jianzhu Liu,* Kun-Ho Song,† Myung-Jo You,§ Dong-Soo Son,¶ Sung-Whan Cho‡ and Duck-Hwan Kim†
*
Laboratory of Veterinary Internal Medicine, College of Animal Science and Veterinary Medicine
Shandong Agricultural University, Taian 271018, China
†
Laboratory of Veterinary Internal Medicine
‡
Laboratory of Veterinary Pathology
College of Veterinary Medicine, Chungnam National University
Daejeon 305-764, Korea
§
College of Veterinary Medicine, Chonbuk National University, Chunju 560-791, Korea
¶
National Livestock Research Institute, R.D.A. Suwon
Gyunggi Province 441-706, Korea

Abstract: The potential recovery effect by oculo-acupuncture (OA) on ethylene glycol-induced

acute renal injury in dogs was investigated. Acute renal damage was induced by ingestion
of ethylene glycol in six mongrel dogs. The dogs were assigned to control (three dogs) and
experimental (three dogs) groups. The control group did not receive any treatment, while the
experimental group was treated with oculo-acupuncture at kidney/urinary bladder region plus
zhong jiao region of the eyes after the induction of renal damage. Serum blood urea nitrogen
(BUN), creatinine, sodium (Na), chloride (Cl), and potassium (K) were measured in both
control and experimental groups. The blood RBC and Hb were also examined. The serum
BUN and creatinine activities in the experimental group were lower than those in the control
group, the serum Na and Cl had the irregular change in both groups, and the blood Hb in the
control and experimental group showed decreasing tendency. Significant differences were
observed on the 3rd and 7th day in BUN, 7th day in creatinine, 2nd day in Na and Cl, and 7th
day in Hb when compared to the control group. Whereas, serum K concentration and RBC in
the experimental group did not change significantly. The recovery findings of the renal injury
were also observed in the experimental group histopathologically.
In conclusion, OA therapy (kidney/urinary bladder region plus zhong jiao region) was
effective for recovery of the renal injury induced by ethylene glycol in dogs.

Keywords: Oculo-Acupuncture; Renal Injury; Ethylene Glycol; Dogs.

Correspondence to: Dr. Duck-Hwan Kim, College of Veterinary Medicine, Chungnam National University, 220
Gung-dong, Yuseong-gu, Daejeon 305-764, Korea. Tel: (+82) 42-821-6756, Fax: (+82) 42-821-8903, E-mail:
dhkim@cnu.ac.kr

241

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 242 J. LIU et al.

Introduction

Acute renal failure (ARF) is the rapid breakdown of renal function that occurs when high
levels of uremic toxins accumulate in the blood. This may result in electrolyte disturbances
and the loss of fluid homeostasis (Thadhani et al., 1996). However, in spite of this rapid
decline in kidney functioning, patients with ARF are often diagnosed with a few other
symptoms (Albright, 2001). In a recent study, 1–5% of patients at hospital admission were
found to have ARF. The condition affects 15–20% of patients in intensive care units, and
involves a 50–70% mortality rate (Singri et al., 2003). Infection and cardiorespiratory
complications are the most common causes of death in patients with ARF. These increases
not only cost of medical care with the long period of hospitalization but also cause patients’
morbidity and mortality (Persson and Patzac, 2005).
Hydrations with saline infusion, N-acetylcysteine, fenoldopam, theophylline,
haemofiltration, and haemodialysis therapy have been used for the treatment of ARF
(Detrenis et al., 2005). However, recently, several papers reported the successful use of
traditional oriental medicine to treat human and animal diseases. Ma (2004) examined the
therapeutic effects of AP on renal damage in 42 cases with gout, and found that AP might
yield good therapeutic effects on early gout complicated with renal damage by adjusting
the metabolism and improving renal function.
Oculo-acupuncture (OA), also known as eye AP in China (Zhao, 1997) but more
accurately called periorbital acupuncture, is a new modality of AP therapy. The therapy
is based on the theory that eyes are closely related to internal organs and meridians, and
the acupuncture applied at the acupoints around the eyes can be used to treat many bodily
diseases. It has 13 special acupoints on eight parts around the orbit of the eye (Zhao, 1997).
OA has been proven to be therapeutic for vomiting (Xiao, 1994; Zhao, 1995; Yang, 2001),
acute muscular sprain (Zhang, 1994; Fu, 1996), apoplexy and hemiplegia (Li, 1995; Ren
and Lin, 2005), and pain (Zhao, 1997; Peng, 1990) in human diseases. Fu et al. (2001)
reported that OA could be effective for the defect of the nerve function in order to treat
acute cerebral infarction. Nie et al. (2003) thought that OA was an ideal therapy for
acute cerebral infarction, and its effect was better than body AP. In addition, Wang et al.
(2004) reported that OA treatment of the liver, heart, and kidney regions could improve
the symptoms of memory disorder and dysmnesia, and could change the ultrastructure of
hippocampus neurons found in the experimental vascular dementia induced in rats. The
therapeutic effects of OA have been studied in various human diseases. However, there is
no report about its therapeutic effect in the veterinary clinical practice.
Hence, in the present study, we aimed to investigate the therapeutic effects of OA on
the ethylene glycol-induced acute renal injury in dogs.

Materials and Methods

Experimental Animals

Six clinically healthy mongrels dogs (two males and four females), aged two to four

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 THE EFFECT OF OCULO-ACUPUNCTURE ON ACUTE RENAL INJURY IN DOGS 243

years and with weight ranging from 2.0 kg to 4.8 kg, were randomly selected for this
study. The animals were kept individually in stainless steel cages, fed with a balanced
diet, and supplied with water ad libitum. The dogs ingested the ethylene glycol, and were
divided into the control group and the experimental group, with three dogs each. As for the
treatment, the control group did not receive any treatment, while the experimental group
was treated with OA at their kidney/urinary bladder region and at the zhong jiao region
of the eye after 24 hours of the renal injury. The present study was performed according
to the rules set for experimental animals by the Ethics Committee of Chungnam National
University.

Induction of Renal Injury

Acute renal damage was induced through the ingestion of ethylene glycol (DC Chemical
Co., Korea, 3 ml/kg, PO).

Oculo-Acupuncture

This study involved needle AP at the kidney/urinary bladder region and the zhong jiao
region of the eyes according to human OA (Zhao, 1997). The 13-mm needle (Hwato®,
Suzhao Medical Appliance Factory, China) was inserted into the kidney/urinary
bladder region (in an oblique direction from TH-23) and the zhong jiao region (in a
horizontal direction from ST01) (Fig. 1). The handle of the needle was slightly pulled
upward or gently twisted with an angle of 10°. The needle was inserted bilaterally
about 13 mm depths, and maintained at the said regions for 20 min once a day for
five days.

Figure 1. Acupoint of oculo-acupuncture (↑ the direction of the needle, TH23: Lateral to the supraorbital
process, where it connects with the orbital ligament; ST01: In the center of the ventral border of the orbit, between
the globe of the eye and the orbital rim).

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 244 J. LIU et al.

Hematological and Blood Chemical Analysis

Blood was collected from the cephalic vein of the dogs in pre, 1, 2, 3, 5 and 7 days after
injury, and was divided into two parts: one (0.5 ml) was collected into a tube containing
EDTA as an anticoagulant for hematological analysis, while another 2 ml sample was
collected into plain tubes with no anticoagulant to provide the serum samples. The
hematological analyses were performed shortly after sampling to examine the changes
in RBC and Hb. The serum was separated by centrifugation at 180 g for 15 min. An
automatic blood chemistry analyzer (SM-4000, B.S Biochemical Systems, Italy) was used
for determining the serum concentration of blood urea nitrogen (BUN), creatinine, sodium
(Na), chloride (Cl) and potassium (K).

Histopathological Examination

Seven days after the renal injury and OA treatment, the dogs were euthanized after the
commencement of the experiment by intravenous injection with pentobarbital sodium
(Nembutal®, Bando Pharmacological Co., Korea, 60 mg/kg, IV), and the kidney tissue
specimen was taken. The tissue specimen was fixed with 10% buffered formalin solution
and processed tissue embedding. Thin paraffin sections were made and stained with
hematoxylin-eosin (H & E) for light microscopy examination.

Statistical Analyses

The significant difference between the control and experimental groups was analyzed
using paired student’s t-test with a database (SPSS v. 12.0, K).

Results

The Change of Hematological Values

RBC was increased after one day of ethylene glycol ingestion, and then it showed a
decreasing tendency in the control group. The change in RBC in the experimental group
showed a similar tendency as the control group. However, there was no significant difference
in RBC between the control group and the experimental group after OA (Fig. 2).
Blood Hb concentrations demonstrated a gradual decreasing tendency after the ingestion
of ethylene glycol in the control group. The same decreasing tendency was found in the
experimental group and the significant difference between the two groups was detected on
the 7th day (p < 0.05) after OA in the experimental group (Fig. 3).

The Change of Blood Chemical Values

Serum BUN concentrations showed a gradual increasing tendency after the ingestion of
ethylene glycol in the control group. Serum BUN concentration in the experimental group

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 THE EFFECT OF OCULO-ACUPUNCTURE ON ACUTE RENAL INJURY IN DOGS 245

was increased after the 3rd day, and then it showed a decreasing tendency, although it had
lower values after the 2nd day than those in the control group. A significant difference was
found on the 3rd day (p < 0.05) and the 7th day (p < 0.01) after OA in the experimental
group compared to the control group (Fig. 4).
Serum creatinine concentrations showed a gradual increase after the ingestion of
ethylene glycol in the control group. Serum creatinine concentrations in the experimental
group demonstrated an increasing tendency until the 2nd day, and then showed a decreasing
tendency at the end of the study. However, those in the experimental group had lower
values after the 2nd day than those in the control group. The significant difference between
the two groups was detected on the 7th day (p < 0.05) after OA in the experimental group
(Fig. 5).
Serum Na concentrations demonstrated irregular changes in the control group after
the ingestion of ethylene glycol. In the experimental group, the concentrations also had
an irregular tendency, but they were lower than those in the control group. The significant

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Figure 3. The change of Hb concentration in the experimental and the control groups.

00478.indd 245 3/21/2007 2:54:01 PM

 246 J. LIU et al.

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Figure 5. The change of creatinine concentration in the experimental and the control groups.

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Figure 6. The change of serum sodium concentration in the experimental and the control groups.

00478.indd 246 3/21/2007 2:54:01 PM

 THE EFFECT OF OCULO-ACUPUNCTURE ON ACUTE RENAL INJURY IN DOGS 247

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Figure 8. The change of potassium concentration in the experimental and the control groups.

difference between the two groups was found on the 2nd day (p < 0.05) after OA in the
experimental group when compare to the control group (Fig. 6).
Serum Cl concentrations demonstrated irregular changes in both the control group and
the experimental group. The significant difference between the two groups was detected on
the 2nd day (p < 0.05) after OA in the experimental group (Fig. 7).
Serum K concentration also demonstrated irregular changes in both groups. No
significant difference was found between the control and the experimental groups
(Fig. 8).

Histopathological Findings

Many oxalate crystals in the tubules and tubular necrosis were observed in the kidney
7 days after the ingestion of ethylene glycol in the control group (Fig. 9). On the other hand,
the regeneration of the tubular epithelium of the kidney was observed in the experimental
group. As compared to those in the control group, the renal injuries in the experimental
group were much improved histopathologically (Fig. 10).

00478.indd 247 3/21/2007 2:54:01 PM

 248 J. LIU et al.

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Figure 9. Oxalate crystals in the tubules and tubular necrosis on the 7th day in the control group (H & E X200).

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Figure 10. Regeneration of the tubular epithelium on the 7th day in the experimental group (H & E X200).

Discussion

In the past few years, many studies have been conducted AP to treat renal injury. In
particular, Che-Yi et al. (2005) reported that AP at the Quchi (LI11) acupoint was an
easy, safe, and effective method of relieving uraemic pruritus. Ma (2004) observed the
therapeutic effects of AP on renal damage in 42 cases with gout complications, and found
that AP might yield good therapeutic effects during the early stage of gout by adjusting
the patients’ metabolism and improving their renal function. Similarly, Tsay et al. (2003)
reported the effectiveness of acupoint massage in improving the quality of life, particularly
the sleeping habits of patients with end-stage renal disease. In addition, Darenkov et al.

00478.indd 248 3/21/2007 2:54:01 PM

 THE EFFECT OF OCULO-ACUPUNCTURE ON ACUTE RENAL INJURY IN DOGS 249

(1993) described that AP could promote the earlier recovery of organs and reduce the
scope of chemotherapy. Thus, he recommended AP for application in urological and
nephrological practice.
Regarding the theory of OA, it is related with the eyes and the meridians. The eyes are
closely related to the internal organs and the meridians, which imply that OA applied at the
acupoints around the eyes can be used to treat many diseases (Zhao, 1997).
The acupoints selected for this study involved the kidney/urinary bladder regions.
Needle insertion was started at TH23-si zhu kong which is the last acupoint in the triple
energizer (TH) meridian, and is related with all the acupoints therein. In addition, the
needle was inserted from ST01-cheng qi which is the first acupoint in the stomach (ST)
meridian in the zhong jiao region, and is related with all the acupoints in the stomach
meridian. As a result, the present OA had some therapeutic effects on renal damage by
means of the stimulation of the TH and ST meridians. However, further research should be
performed for the validation of this finding.
As for the biochemical parameters of the present study, the results of OA demonstrated
that concentrations of BUN, creatinine, Na, Cl, and Hb in serum in the experimental group
were significantly different from those in the control group. These changes were associated
with the recovery of the kidney tissues by OA in the kidney/urinary bladder and stomach
regions.
On the other hand, the histopathological features of the renal tissues in ethylene glycol
intoxication revealed that an oxalate crystal in the tubules and tubular necrosis was found
on the 7th day in the control group, while a mild oxalate crystal of the kidney was observed
in the experimental group. The recovery findings for renal injury were significantly
different, histopathologically, in the experimental group. It was therefore assumed that the
histopathological recovery finding for the injured kidney tissue backed-up the therapeutic
effect of OA.
In this study, we investigated the therapeutic effect of OA on dogs with acute renal
injury induced by ethylene glycol. It was demonstrated that OA was effective in the
recovery of dogs’ damaged renal tissues.

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