Current Eye Research

ISSN: 0271-3683 (Print) 1460-2202 (Online) Journal homepage: http://www.tandfonline.com/loi/icey20

Correlations Between MMPs and TIMPs Levels in

Aqueous Humor from High Myopia and Cataract
Patients

Yan Jia, Dan-Ning Hu, Jing Sun & Jibo Zhou

To cite this article: Yan Jia, Dan-Ning Hu, Jing Sun & Jibo Zhou (2016): Correlations Between
MMPs and TIMPs Levels in Aqueous Humor from High Myopia and Cataract Patients, Current
Eye Research, DOI: 10.1080/02713683.2016.1223317

To link to this article: http://dx.doi.org/10.1080/02713683.2016.1223317

Published online: 19 Oct 2016.

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CURRENT EYE RESEARCH
http://dx.doi.org/10.1080/02713683.2016.1223317

SHORT COMMUNICATION

Correlations Between MMPs and TIMPs Levels in Aqueous Humor from High Myopia
and Cataract Patients
Yan Jiaa,*, Dan-Ning Hub,*, Jing Suna, and Jibo Zhoua
a
Department of Ophthalmology, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University, School of Medicine, Shanghai, P. R. China;
b
Departments of Ophthalmology and Pathology, New York Eye and Ear Infirmary of Mount Sinai, New York, NY USA

ABSTRACT ARTICLE HISTORY

Purpose: To study the relationships between matrix metalloproteinases (MMP)-2, MMP-3, and tissue Received 25 February 2016
inhibitors of matrix metalloproteinases (TIMP)-1, TIMP-2, and TIMP-3 aqueous humor levels in patients Revised 17 July 2016
with high myopia or cataract. Accepted 2 August 2016
Materials and Methods: MMPs and TIMPs protein levels in 65 aqueous humor samples collected from KEYWORDS
patients with high myopia or cataract during cataract or clear lens extraction surgery were measured Aqueous humor; correlation
with the Luminex xMAP Technology. The relationship between MMPs and TIMPs levels was analyzed study; MMPs; myopia; TIMPs
with Spearman’s correlation test.
Results: MMP-2 levels, but not MMP-3 levels, were increased in the aqueous humor from high-myopia
patients. Levels of TIMP-1, -2, and -3 were positively and very significantly correlated with the MMP-2 levels
(TIMP-1: r=0.626, p < 0.001; TIMP-2: r = 0.545, p < 0.001; TIMP-3: r = 0.439, p < 0.001). TIMP-2 and-3 levels did
not significantly correlate with MMP-3 levels (TIMP-2: r = 0.175, p > 0.05; TIMP-3: r = 0.127, p > 0.05) and TIMP-
1 levels only marginally correlated with MMP-3 levels (r = 0.278, 0.01< P < 0.05).
Conclusions: Compared to the present findings with the relationship of MMPs and TIMPs in other fields
of medicine, our results are consistent with the homeostasis hypothesis that the increase of TIMPs serves
as a compensation reaction to inhibit the excessive degradation caused by the increase of MMPs and
limits the development of myopia.

Introduction Changes in TIMPs levels during the development stage of

myopia varied in different reports and the relationship
During the development stage of myopia, there is a loss of
between MMPs and TIMPs was conflicted.2–10
extracellular matrix (ECM) in the mammals sclera, which
To our best knowledge, the correlation between MMPs and
causes a weakness of the sclera and results in elongation of
TIMPs levels in aqueous humor from myopic patients has not
the axial length (AL) and the development of myopia.1 In
been reported previously. Recently, we measured the MMPs
experimental animal studies, this process has been linked with
and TIMPs levels in the aqueous humor from 65 patients with
the increase of matrix metalloproteinases (MMPs) levels in
high myopia or cataract and found that the MMPs and TIMPs
the sclera.2–9
levels in high-myopia patients were significantly different
MMPs are a group of enzymes that can degrade virtually
from those in cataract patients.36
all ECM components. The activity of these proteases is tightly
The purposes of the present study were to analyze the
regulated by specific inhibitors, known as TIMPs.10–17 During
correlation between MMPs and TIMPs levels in the aqueous
various processes involving the degradation of ECM, the
humor from these patients, compared with the results
relationship between the changes of MMPs and TIMPs levels
obtained from experimental animal myopia studies and to
is complicated and even contradictory (TIMPs increase with
evaluate the previous hypotheses regarding the relationship
MMPs).12,14–36 There are two different hypotheses for the
between MMPs and TIMPs.
explanation of these contradictory changes: 1) the homeosta-
sis hypothesis: the increase of TIMPs reflects a cellular com-
pensatory reaction to counteract and limit the intensive Materials and methods
degradation of ECM by MMPs;15–17,27,28,33,34 and 2) the
Patients and samples
MMPs-independent hypothesis: TIMPs can directly affect
the cells’ behaviors, which are independent of the inhibition The subjects and methods for the measurement of various
of MMPs activities.15–17,29,30,33,34 factors used in this study have been reported previously.36
In an experimental myopia model, MMPs levels, especially Briefly, aqueous samples were collected from 65 patients dur-
MMP-2, are increased in the sclera in myopic animals.2–9 ing cataract or clear lens extraction surgery. The aqueous

CONTACT Jibo Zhou; Jing Sun zhoujibo1000@aliyun.com; sophiasj@126.com Department of Ophthalmology, Shanghai Ninth People’s Hospital, Shanghai
Jiaotong University, School of Medicine, 639 Zhizaoju Road, Shanghai 200011, P. R. China.
*The first two authors contributed equally to this work.
© 2016 Taylor & Francis
2 Y. JIA ET AL.
humor (0.1–0.2 ml) was collected at the beginning of opera-
tion and stored under −80°C. The protein levels of MMP-1,
MMP-2, MMP-3, TIMP-1, TIMP-2, and TIMP-3 were mea-
sured with the Luminex system (Luminex xMap Technology,
Bio-Rad, Hercules, CA, USA) by using commercially available
Milliplex xMAP Kits (Millipore Corporation, Billerica, MA,
USA).36 All of these factors could be detected in the aqueous
humor, with the exception of MMP-1. Therefore, the data of
MMP-2, MMP-3, and TIMPs levels have been used in the
present study for investigating the relationships between these
factors.36
This study was performed in accordance with the
Declaration of Helsinki. All patients signed an informed con-
sent and the Institutional Review Board at the Shanghai Ninth
People’s Hospital approved the study.36
Analysis of relationships between MMPs and TIMPs
The relationship between the levels of various TIMPs and
MMPs was analyzed separately by testing the significance of
correlation coefficient between various groups.
Statistics
The original data were not normally distributed by the
Kolmogorov–Smirnov test. Therefore, the results were
expressed as median and range interquartile range (25th and
75th percentiles) in continuous variables that are not normally
distributed.36 The relationship between MMPs and TIMPs was
analyzed with Spearman’s correlation test by using SPSS 22.0
software PSS, Inc. (Chicago, IL, USA). A two-tailed p < 0.05
was considered to be statistically significant.
Results
In our previous study, the patients were separated into three
groups: Group A with normal AL (≤ 24 mm) and mean +/-
standard deviation refractive error at −0.8 ± 2.1 D; Group B with
elongation of AL (24–26 mm) and high myopia (−9.3 ± 5.3 D);
and Group C with excessive elongation of AL (≥26 mm) and very
high myopia (−17.6 ± 6.8 D).36 The levels of MMP-2 in Group C
were significantly higher than those in Group A.36 TIMPs levels
were the highest in Group C and the lowest in Group A.36
In the present study, all TIMPs levels were positively and
very significantly correlated with MMP-2 levels [TIMP-1: r =
0.626, p < 0.001 (Figure 1a); TIMP-2: r = 0.545, p < 0.001
(Figure 1b); TIMP-3: r = 0.439, p < 0.001 (Figure 1c)].
In our previous report, no significant difference of MMP-3
levels in the aqueous humor could be found in the three
different groups (p > 0.05).35 In this study, the correlation
coefficient between MMP-3 and TIMP-1 levels was 0.278,
which was only moderately significant (p = 0.025). No sig-
nificant correlation was found between the levels of MMP-3
and TIMP-2 (r = 0.175, p > 0.05) or TIMP-3 (r = 0.127,
p > 0.05).
Table 1 shows the correlation between the aqueous humor
protein levels of MMPs and TIMPs in cataract and high-
myopia patients. Only one of the TIMPs levels was correlated
with MMP-3 in both cataract and high-myopia patients. All
TIMPs levels were significantly correlated with MMP-2 levels
in high-myopia patients, but only TIMP-1 and -3 levels were
correlated with MMP-2 levels in cataract patients, suggesting
that the relationship between MMPs and TIMPs is slightly
stronger in high myopia than in cataract patients.
Discussion
The relationship between MMP-2 and TIMPs levels during
the development stage of myopia is varied and can be classi-
fied into three categories2–10:
(1) Positive correlation: MMP-2 activities and TIMPs
RNA and protein levels increased in myopia (tree
shrew).2,3
(2) Negative correlation: MMP-2 RNA levels increased
and TIMPs levels decreased in myopia (chicken fibrous
sclera).5
(3) Others: Norton’s team reported that scleral TIMP-1
and -3 RNAs levels decreased, but MMP-2 levels did
not change in form deprivation myopia (FDM) and
lens-induced myopia (tree shrew).10 Shaeffel’s team
reported that MMP-2 RNA levels increased in chicken
fibrous sclera.9
During the recovery stage of experimental myopia, MMP-2
levels were decreased and TIMPs levels usually increased.5–7
Therefore, the relationships between TIMPs and MMP-2 in
experiment animal myopia are very complicate and depend
on various factors.2–10 In the present study, the myopia was in
the stationary stage, and hence it is difficult to compare with
the results from experimental animals directly. The results in
our study suggested that the levels of MMP-2 and various
TIMPs were increased.
MMP-3 levels in the aqueous humor were mainly not
correlated to TIMPs levels. This is consistent with the results
from experimental animal studies that scleral MMP-3 mRNA
levels did not significantly change in FDM or defocus
myopia.6–8
In the study of the relationship of MMPs and TIMPs in
physiological or pathological processes other than myopia, the
elevation of MMPs levels is frequently associated with the
increase of TIMPs in various physiological changes related
to endocrine and metabolism18,19 and in pathological
changes.12,14–17,20–32 In the field of oncology,12,15–17,25–33
MMPs degrade ECM and release growth factors from ECM,
which promotes angiogenesis and stimulates the migration
and invasion of cancer cells.12,14–17 The expression and acti-
vation of MMPs are increased in almost all human cancers
compared to that of normal tissues. Positive correlations have
been found between MMPs expression and poor prognosis in
many types of cancer.12,14–17 TIMP levels in the serum or
cancer cells often up-regulated, instead of down-regulated,
in many cancers.12,14–17,25–32
Homeostasis hypothesis has been developed for the expla-
nation of this paradoxical relationship.16–18,29,34,35 Increased
TIMPs expression may be a reflection of cellular response to
the increased MMPs expression. This can counteract and
inhibit MMPs-mediated ECM degradation and the progress
 CURRENT EYE RESEARCH 3

Figure 1. Correlation between aqueous MMP-2 and TIMP-1, -2, and -3 levels. MMPs/TIMPs levels in the aqueous humor from 65 patients with myopia or cataract
during cataract or clear lens extraction surgery were measured with the Luminex xMAP Technology by using commercially available Milliplex xMAP Kits and analyzed
by the Spearman correlation test. a. Correlation between TIMP-1 and MMP-2. TIMP-1 levels were positively and significantly correlated with MMP-2 levels (r = 0.626,
p < 0.001). b. Correlation between TIMP-2 and MMP-2. TIMP-2 levels were positively and significantly correlated with MMP-2 levels (r =0.545, p < 0.001). c.
Correlation between TIMP-3 and MMP-2. TIMP-3 levels were positively and significantly correlated with MMP-2 levels (r = 0.439, p < 0.001). d. Correlation between
TIMP-1 and MMP-2 (three outliers have been removed). TIMP-1 levels were positively correlated with MMP-2 levels (r = 0.573, p < 0.001).

Table 1. Correlation between aqueous humor protein levels of MMPs and TIMPs
in cataract and high-myopia patients.
Cataract (n = 29) High myopia (n = 36)
poor prognosis,12,15–17,29–33,35 indicating that TIMPs might
have a direct effect on the cancer cells (MMPs-independent
r p r p
hypothesis). There is little information available on the
MMP-2 TIMP-1 0.532 0.003* 0.626 <0.001*
TIMP-2 0.197 0.306 0.555 <0.001* MMPs-independent effects of TIMPs that related to the devel-
TIMP-3 0.530 0.003* 0.353 0.034* opment of myopia. Further study of this effect of TIMPs may
MMP-3 TIMP-1 0.224 0.242 0.448 0.006*
TIMP-2 −0.406 0.029* 0.311 0.065 be useful for understanding the roles of TIMPs in the devel-
TIMP-3 0.049 0.803 0.312 0.064 opment of myopia.
High myopia: <−6.0 D; r: correlation coefficient; p: p-value, * p < 0.05. In conclusion, comparing the present findings with the
relationship of MMPs and TIMPs in other fields of biology
and medicine, it seems that our results are consistent with the
of the cancer, which represents a homeostatic mechanism to homeostasis hypothesis that the increase of TIMPs serves as a
limit the damage caused by the excessive up-regulation of compensation reaction to inhibit the excessive degradation
MMPs.15–17,28,33,34 and to limit the development of myopia caused by an increase
The present study revealed that TIMPs levels increased in of MMPs.
myopia and had a positive significant correlation with MMP-2
levels in myopic eyes. This relationship is slightly stronger in
high-myopia than in cataract patients. Our result is consistent Declaration of interest
with the homeostasis hypothesis that the up-regulation of
TIMPs may be a cellular reaction to the excessive degradation All authors have no financial interest in the publication of this article.
of ECM caused by the increase of MMPs levels in myopia.
However, the causal-result relationship between the
Funding
increase of MMP-2 and TIMPs levels still requires further
studies. This work was supported by the National Nature Science Foundation of
It has been reported that in several cancers, the increase of China, (Foundation Number: 81371050) and Shanghai Municipal
Commission of Health and Family Planning Found (Foundation
TIMPs levels in the blood or tumor tissues is associated with a Number: 201440354).
4 Y. JIA ET AL.
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