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com J Tradit Chin Med 2012 September 15; 32(3): 477-481
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Influence of catgut implantation at acupoints on leptin and insulin
resistance in simple obesity rats
Runhu Yan, Xiaoming Liu, Jie Bai, Jiang Yu, Jinsong Gu
aa
Runhu Yan, College of Cosmetology Medicine of Dalian
Medical University; Research Institute of Integrated Tradi-
tional and Western Medicine of Dalian Medical University,
Dalian 116044, China
Xiaoming Liu, Dermatology of The First Affiliated Hospital
of Dalian Medical University, Dalian 116011, China
Jie Bai, Jiang Yu, Jinsong Gu, College of Cosmetology Med-
icine of Dalian Medical University, Dalian 116044, China
Supported by Science Research Projects of Colleges and
Universities of Liaoning Provincial Committee of Education
(Project number: 05L121)
Correspondence to: Xiaoming Liu, Dermatology Depart-
ment of the First Affiliated Hospital of Dalian Medical Univer-
sity, NO.222 Zhongshan Road, Dalian 116011, China. lxm_
1956@yahoo.com.cn
Telephone: +86-411-83635963-3087
Accepted: June 27, 2012
Abstract
OBJECTIVES: To investigate the influence of catgut
implantation at acupoints on leptin resistance (LR)
and insulin resistance (IR) in the simple obesity rat.
METHODS: Rats were made obese with high-fat di-
ets, after which surgical catgut was implanted at
Zusanli (ST 36) and Neiting (ST 44) acupoints once
a week for 4 weeks (implantation group). Rats from
the implantation group were compared with nor-
mal rats and unoperated obese rats (control
group). Gene expression of the leptin receptor
(OB-R) was evaluated using RT-PCR and northern
blot. Serum and hypothalamus leptin and insulin
(INS) levels were determined by radioimmunoas-
say. Body weight, Lee's index, body fat, serum and
hypothalamus leptin and INS levels, and hypotha-
lamic OB-R gene expression were determined be-
fore and after treatment.
JTCM | www. journaltcm. com
ISSN 0255-2922
© 2012 JTCM. All rights reserved.
BASIC INVESTIGATION
RESULTS: Body weights, Lee's index, body fat, and
serum leptin and INS levels were significantly high-
er in obese than in normal rats. Hypothalamic
leptin and INS levels and OB-R gene expression
were significantly lower in obese rats. Catgut im-
plantation at acupoint promoted weight loss and
decreased serum leptin and INS levels. Hypotha-
lamic leptin and INS levels and OB-R gene expres-
sion increased significantly.
CONCLUSIONS: Catgut implantation at acupoint
adjusts central and peripheral leptin and promotes
hypothalamic OB-R gene expression. This may be
an important method for regulation of LR, IR and
abnormal endocrinology and metabolism.
© 2012 JTCM. All rights reserved.
Key words: Thread-embedding acupoint ligation;
Obesity; Leptin resistance; Insulin resistance; Recep-
tors, Leptin; Blood-cerebral barrier
INTRODUCTION
Simple obesity is one of the most serious epidemics
around the world, and is receiving ever-increasing at-
tention from the medical world. Obesity not only af-
fects the shape of the body and physical appearance, it
also increases the incidence of cardiovascular disease, di-
abetes, hyperlipidemia, infertility and tumors.1 Addi-
tionally, it severely influences physical and mental
health, weakens immunity and increases the mortality
rate.2 At present, obesity has taken the place of malnu-
trition and infectious diseases and become one of the
most important human health hazards. As obesity has
become a global health problem in the 21st century,
the study on obesity has been a central focus in medi-
477 September 15, 2012 | volume 32 | Issue 3 |
Yan RH et al. Influence of catgut implantation at acupoints in simple obesity rats
cal circles.3 Modern research shows that most obese pa-
tients have insulin resistance (IR) and leptin resistance
(LR).4,5 There are very few reports on experimental
studies using catgut implantation at acupoint for the
treatment of obesity-related IR and LR. Therefore, this
study aims to evaluate the effects of catgut implanta-
tion at acupoints on body weight and LR and IR in
obese rats.
MATERIALS AND METHODS
Animals and grouping
One-hundred one-month-old newly weaned male
Sprague-Dawley rats, body weight 50-70 g, were pro-
vided by the Center for Experimental Animals of Da-
lian Medical University. Body length, body weight and
Lee's index were determined periodically. After 12 weeks
of feeding on a high-fat diet, rats with body weights
surpassing those of normally-fed rats by 20% or more
were determined to be experimentally obese. A low
number of high-fat fed rats were excluded, because
their body weights increased slowly and were similar to
normally-fed rats, indicating obesity resistance.
Rats fed normally were categorized as the normal
group (n=20). Rats in which experimental obesity was
successfully induced (n=43) were randomly divided in-
to two groups. One group received no further treat-
ment (control group; n=21). The other group was treat-
ed by catgut implantation at acupoint (implantation
group; n=22). Twelve rats were sampled randomly
from each of the groups.
Treatment method
Rats in the implantation group were put into a rat fixa-
tion machine and the Zusanli (ST 36) and Neiting (ST
44) acupoints were selected. The implantation areas
were sterilized. No. 2 surgical catgut of different
lengths was inserted at both acupoints according to
muscle abundance degrees, and fixed with adhesive
plaster. The catgut was implanted once a week between
8:00 am and 9:00 am for 4 weeks, and acupoints on
both sides were used alternatively.
Experimental methods
Rats in all groups were fed the ordinary full valence
fodder during the experimental period, with free access
Table 1 Comparison of body weight, body length and Lee's index among the groups
Group Sample number (n=12) Weight (g)
B 384.2±33.7
Normal
A 396.5±28.8
B 517.0±29.4
Control
A 513.8±29.1a
B 516.7±28.4a
Implantation
A 440.2±26.2 bcd
Notes: B means before treatment; A means after treatment; bP<0.01, comparison between B and A within the same group; cP<0.05, aP<
0.01, comparison between the control group and the normal group (B∶B or A∶A); dP<0.01, comparison between the implantation group
and the control group (B∶B or A∶A) (same below).
JTCM | www. journaltcm. com 478
to food and water. Fodders and water were replaced ev-
ery day. The rats in the implantation group were treat-
ed by catgut implantation at acupoint. The rats in the
normal group and the control group were put into the
rat fixation machine for 15 min once a week for 4
weeks. Food and water intakes, production of urine
and feces, body weight, body length and Lee's index
were determined weekly. After the experiment the rats
were fasted overnight and decapitated the next day.
Whole blood was collected and separated, and the se-
rum was kept at -20°C until analysis. Pericardial, peri-
nephric and paradidymis fat was rapidly separated and
weighted. At the same time, whole brains were rapidly
separated and kept in liquid nitrogen. The brains of 6
rats per group were thawed the following day. The hy-
pothalami were separated, weighed and fully homoge-
nized in glass homogenate tubes containing 1 mL
HC1 (1 mol/L) in an ice bath. Following homogeniza-
tion, samples were kept at room temperature for 100
min and centrifuged at 4000 r/min for 20 min at 4°C.
To neutralize the acid, 1 mL NaOH (1 mol/L) was add-
ed to the supernatant. Neutralized samples were centri-
fuged at 4000 r/min for 10 min and the supernatant
(hypothalamus homogenate) was kept at -20° C until
measurement of the leptin and insulin concentrations.
The brains of the other 6 rats in each group were taken
from the liquid nitrogen, and the hypothalami were
separated and weighed before RNA extraction.
Test methods
The central lab of Dalian Medical University adopted
RT-PCR and northern blot to evaluate the expression
of the leptin receptor (OB-R) gene, and radioimmuno-
assay to determine the serum and hypothalamic leptin
and INS content.
Statistical methods
The SPSS12.0 software was used to analyze the data.
Data are presented as mean±SD. Student's t-tests were
used for comparison between groups, and multiple re-
gression analysis was adopted for the relevant analysis.
RESULTS
The influence of catgut implantation at acupoint on
body weight, body length and Lee's index in obese rats
Length (cm) Lee's index
24.4±0.8 298.5±4.0
24.5±0.7 301.0±4.2
a
25.1±0.5 319.5±3.6a
25.4±0.5 317.3±3.8a
25.2±0.5 320.0±4.4a
25.4±0.4 298.4±4.0bd
September 15, 2012 | volume 32 | Issue 3 |
 Yan RH et al. Influence of catgut implantation at acupoints in simple obesity rats

Table 1 shows that body weight and Lee's index were
significantly higher in the obese rats than in the nor-
mal rats. After treatment, body weight and Lee's index
decreased significantly.
The effects of catgut implantation at acupoint on fat
mass in different parts of the body in obese rats
The fat mass in the control group was significantly
higher than in the normal group. Fat mass in the im-
plantation group was significantly lower than in the
control group, suggesting that catgut implantation at
acupoint induces weight loss (Table 2).
The influence of catgut implantation at acupoint on
the hypothalamic and serum leptin content in obese
rats
The serum leptin content in the control group was sig-
nificantly higher than in the normal group. The hypo-
thalamic leptin content and the hypothalamus/serum
leptin ratio were significantly lower in the control
group than in the normal group. Compared with the
control group, the serum leptin content in the implan-
tation group had decreased significantly, while the hy-
pothalamic leptin content and the hypothalamus/se-
rum leptin ratio had increased significantly (Table 3).
Table 2 Comparison of fat mass in different parts of the body among the groups (n=12)
Group Pericardial fat (g)
Normal 0.54±0.23
Control 0.84±0.14
Implantation 0.55±0.09
The influence of catgut implantation at acupoint on
the hypothalamic and serum insulin content
The serum INS content in the control group was sig-
nificantly increased in the gray value compared with
the normal group. The hypothalamic INS content and
hypothalamus/serum INS ratio were significantly de-
creased. Compared with the control group, the serum
INS content in the implantation group was significant-
ly decreased, while the hypothalamic INS content and
INS ratio were significantly increased (Table 4).
RT-PCR analysis of the gene expression of
hypothalamic OB-R
The normal group, gene expression of hypothalamic
OB-R in the control group was significantly decreased.
Gene expression of hypothalamic OB-R was signifi-
cantly increased in the implantation group compared
with the control group (Table 5).
Northern blot analysis of the gene expression of
hypothalamic OB-R
Normal group, gene expression of hypothalamic OB-R
in the control group was significantly decreased in den-
sity value. Gene expression of hypothalamic OB-R in
the implantation group was increased significantly
compared with the normal group.
Perirenal fat (g) Paradidymis fat (g)
3.53±2.16 2.65±1.01
7.62±2.62 5.47±1.81
4.30±0.75 3.14±0.82

Table 3 Comparison of the hypothalamic and serum leptin content among the groups
Hypotha Serum leptin
Group Hypothalamus leptin/serum leptin (n=6)
lamus leptin (ng/mg) (n=6) (ng/mL) (n=12)
Normal 0.440±0.120 13.820±2.690 0.033±0.016
Control 0.220±0.140 17.670±3.330 0.012±0.004
Implantation 0.670±0.230 14.310±1.810 0.036±0.013

Table 4 Comparison of the hypothalamic and serum insulin (INS) content among the groups
Hypothalamus INS Serum INS Hypothalamus leptin/serum leptin (n=
Group
(μIU/ mg) (n=6) (μIU/ mL) (n=12) 6)
Normal 1.790±0.230 92.010±41.260 0. 020±0.008
Control 1.020±0.580 175.160±72.320 0. 007±0.004
Implantation 2.650±0.800 77.320±25.170 0. 033±0.017

Table 5 Comparison of the gene expression of hypothalamic OB-R among the groups
Group (n) OB-R (gray value) β-actin (gray value) OB-R/β-actin
Normal (5) 19.95±1.52 96.57±21.82 0.21±0.03
Control (5) 18.83±3.40 141.34±13.67 0.13±0.02
Implantation (6) 22.87±3.37 121.46±24.48 0.18±0.02

JTCM | www. journaltcm. com 479 September 15, 2012 | volume 32 | Issue 3 |
Yan RH et al. Influence of catgut implantation at acupoints in simple obesity rats
Table 6 Northern blot analysis of the gene expression of hypothalamic OB-R ( xˉ ±s)
Group (n=6) OB-R (density value)
Normal 1087.830±281.180
Control 1037.320±106.500
Implantation 1726.670±518.550
DISCUSSION
According to traditional Chinese medicine, simple obe-
sity is caused by dysfunction of the spleen and stom-
ach. The whole process of the treatment of catgut im-
plantation at acupoint includes point-blocking therapy,
stylostixis, pricking blood therapy, histotherapy and
cutting therapy. It also includes the needle implanting
effect and after-effect.6 Catgut implantation at acu-
point improves and extends acupuncture therapy.7 Tra-
ditional acupuncture cannot compete with its durative
effect. The treatment of catgut implantation at acu-
point is only applied once every two weeks, which
saves time and has fewer needle insertions and thus less
pain for each treatment. Therefore, patients have high
acceptance of the treatment, which can also easily be
fitted into the high-speed and high-efficiency lifestyles
of modern people.
The weight loss treatment of catgut implantation at
acupoint not only reduces body weight and fat, but al-
so improves related complications, improving the gen-
eral health of the patient. Other advantages are that it
is simple, convenient and effective and has no toxic
side effects, making a weight loss method worth being
popularized.
Research on the causes and pathogenesis of obesity us-
ing molecular biotechnology has achieved great success-
es. In 1994, Zhang8 and others successfully cloned the
genes relevant to obesity in rats, and identified the cor-
responding human obesity gene and its protein prod-
uct leptin, a hormone secreted by lipocytes with many
different roles. Most importantly, leptin influences the
hypothalamic centers that regulate body weight, result-
ing in low appetite, high energy consumption and
weight reduction. It has been proven that leptin, bind-
ing with OB-R in the central nervous system,9 can sup-
press the increase in NPY10 and reinforce the decrease
in α-MSH, r-MSH, β-EP, etc. to regulate food intake
and energy metabolism.11 The main way in which
leptin participates in the regulation of food intake regu-
late energy metabolism is by stimulating the synthesis
and release of NPY through inhibiting hypothalamic
nuclei arcuati NPY nerve fibers after binding to OB-R
in the central nervous system.12
Studies in recent years have suggested that there may
be an endocrine lipid-insulin axis between adipose tis-
sue and pancreatic islets, which constitutes a functional
feedback loop through leptin and INS. In normal cir-
cumstances, leptin inhibits the secretion of insulin by
biding with OB-R in the β-cells of the pancreatic is-
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β-actin (density value) OB-R/β-actin(density ratio)
47266.230±15122.760 0.023±0.002
70401.030±17428.600 0.013±0.002
48928.000±9344.060 0.036±0.011
lets.13 In obese patients there may be a disorder in the
endocrine lipid-insulin axis because of the insensitivity
of β-cell OB-R to leptin. This means that leptin can-
not effectively inhibit the secretion of INS by the
β-cells, leading to hyperinsulinemia.14 Hyperinsu-
linemia and the insensitivity to leptin cause a positive
feedback loop that promotes synthesis of fat and aggra-
vates hyperleptinemia.15
Clinical observations have shown that patients with
simple obesity have hyperleptinemia and hyperinsu-
linemia compared with non-obese people, which is sug-
gestive of LR and IR.16 Our experimental results show
that the serum leptin and INS levels in the obese rats
were similar to those of patients with simple obesity.
These results demonstrate that obese organisms show
characteristics of LR and IR. In short, obstruction of
the pathway for transportation of leptin from the
blood to brain and impaired OB-R gene expression in-
crease NPY secretion by the hypothalamus, which re-
sults in increased food intake and decreased energy con-
sumption, hyperinsulinemia, increased fat deposition,
and IR. In obese patients, leptin insensitivity of β-cell
OB-R results in ineffective inhibition of INS secretion
by leptin, which further aggravates hyperinsulinemia
and LR. This vicious cycle will inevitably result in the
aggravation of obesity.
Our research shows that body weight, Lee's index and
the body fat content decreased significantly after catgut
implantation at acupoint in obese rats, indicating that
this treatment has a benign weight loss effect. Serum
leptin and INS concentrations decreased significantly,
while the hypothalamic leptin and INS content, the ra-
tio of leptin in the brain to blood INS and the gene ex-
pression of hypothalamic OB-R increased significantly.
We demonstrated that catgut implantation at acupoint
decreased peripheral leptin and INS in obese organ-
isms, increasing leptin and INS and OB-R gene expres-
sion of OB-R in the central nervous system, which can
correct LR and IR.
In sum, pathway obstruction for transportation of
leptin and INS from the blood to brain and impaired
gene expression of OB-R and the insulin receptor may
explain LR, IR and obesity. The moderate regulating ef-
fect of catgut implantation at acupoint on central and
peripheral leptin and INS levels and its promotion of
the gene expression of hypothalamic OB-R in obese or-
ganisms may be an important method for correcting
LR, IR and abnormal endocrinology and metabolism,
making catgut implantation at acupoint an important
potential strategy for weight reduction.
480 September 15, 2012 | volume 32 | Issue 3 |
 Yan RH et al. Influence of catgut implantation at acupoints in simple obesity rats
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