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28 Scheller, A. et al. (1999) Structural requirements for cellular uptake
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Nat. Biotech. 16, 857–861

Reseptor teraktivasi-protein: penjaga untuk

peradangan?
Tom M. Cocks and James D. Moffatt

Reseptor teraktivasi protease sel (PAR) tampaknya telah berevolusi untuk mendeteksi protease serin aktif
enzimatik ekstraseluler seperti trypsin dan trombin. Lokasi dominan PAR pada endotelia dan epitel dan
penemuan enzim seperti trypsin dalam jaringan ini, bersama dengan hubungan PARs dengan jalur sitoprotektif,
memberikan informasi baru tentang pensinyalan autokrin dan paracrine dalam hambatan kritis ini. Dalam artikel
ini, cara-cara di mana distribusi dan fungsi PAR dapat dimanfaatkan oleh para farmakologis sebagai strategi baru
terapi antiinflamasi dibahas.

Trombin enzim yang ditanggung plasma tidak hanya sedemikian rupa sehingga pengembangan terapi efektif mendeteksi dan
berkontribusi langsung pada koagulasi dengan mengkatalisis berdasarkan PAR mungkin menjadi tugas yang menantang. menanggapi
pembentukan fibrin, tetapi juga mengatur efek seluler seperti Seperti namanya, PAR bergantung pada aktivitas proteolitik trombin
aktivasi trombosit dan vasodilatasi1. Beberapa efek trombin enzim untuk memulai pensinyalan sel. Dengan demikian, ekstraseluler yang
sekarang diketahui dimediasi oleh anggota keluarga reseptor PARs memiliki situs pembelahan spesifik untuk trombin, aktif secara
teraktivasi protease (PAR) yang muncul1,2. Penemuan reseptor trypsin dan kemungkinan protease serin lainnya dalam enzimatis dan
unik pertama ini, yang ditunjuk PAR1, dan penjelasan domain N-terminal ekstraselulernya (Gbr. 1). Setelah enzim tryptic
mekanisme baru aktivasi telah membuka bidang penelitian yang pembelahan pada situs-situs spesifik ini, terminal-N yang tertentu. Empat
menarik yang menggabungkan unsur-unsur biokimia enzim sekarang baru untuk masing-masing fungsi reseptor berfungsi PAR (PAR1, PAR2,
tradisional dengan farmakologi reseptor yang tidak sebagai 'ligan tertambat' dan berikatan secara intramolekul ke PAR3,
konvensional1,2. Meskipun pemahaman kita tentang biologi suatu lokasi, mungkin pada loop ekstraseluler kedua, untuk
molekuler PARs berkembang pesat, peran fisiologis PAR - memulai penggandaan G-protein dan pensinyalan sel1, 2.
T.M. Cocks,
dengan pengecualian aktivasi trombosit - sebagian besar tetap Mekanisme autoaktivasi yang unik ini memisahkan PAR dari NHMRC Senior
tidak diketahui. Penelitian tentang fungsionalitas PARs telah reseptor protease lain seperti yang untuk aktivator Research Fellow,
didorong oleh keyakinan bahwa PARs berkontribusi pada plasminogen dan faktor Xa [yaitu: effector cell protease Department of
ketidaknyamanan yang terkait dengan peradangan kronis. receptor 1 (EPR-1)], yang mengikat enzim-enzim ini di daerah- Pharmacology,
Tinjauan ini mengeksplorasi pandangan yang lebih umum dari daerah yang jauh dari situs katalitik mereka8,9. Oleh karena The University of
Melbourne, Parkville,
fungsionalitas PAR dan menyarankan bahwa protease yang itu, PAR dapat dianggap sebagai 'sensor' aktivitas proteolitik
Victoria 3010,
mengaktifkan PARs mungkin memiliki peran penting dalam daripada reseptor dalam arti tradisional. Anehnya mungkin, Australia.
perilaku proinflamasi antiinflamasi dan kronis kronis dari mimetik peptida sintetik dari urutan ligan tertambat PAR juga E-mail: t.cocks@
hambatan pertahanan tubuh, endotelia dan epitel. Perbedaan dapat mengaktifkan PAR secara terpisah dari kebutuhan akan pharmacology.unimelb.
mendasar antara efek potensial akut dan kronis dari aktivasi proteolisis (Gbr. 1). Namun, tanpa bukti aktivator peptida edu.au
PAR ini mencerminkan sifat 'peradangan pedang bermata dua', endogen, peran utama PAR tampaknya adalah untuk
and
J.D. Moffatt,

0165-6147/00/$ – see front matter © 2000 Elsevier Science Ltd. All rights reserved. PII: S0165- TiPS – March 2000 (Vol. 21) 1
6147(99)01440-6
 REVIEW
Research Officer, Department of Cardiac Surgery, Royal Melbourne Hospital,
Parkville Victoria 3051, Australia and Department of Pharmacology,
The University of Melbourne, Parkville, Victoria 3010, Australia.
E-mail: j.moffatt@ pharmacology.unimelb. edu.au

2 TiPS – March 2000 (Vol. 21)

dan PAR4, dalam urutan kronologis kloning pada
manusia1) diketahui (Tabel 1). Semua reseptor ini
diaktifkan oleh trombin, trypsin atau keduanya. Meskipun
protease serin lain seperti tryptase sel mast dan faktor Xa
dapat mengaktifkan PAR, sejauh ini tidak ada PAR baru
yang ditemukan yang tidak diaktifkan oleh trombin atau
enzim mirip trypsin. Oleh karena itu, mudah bagi
farmakologis untuk mengkarakterisasi PARs dengan
sensitivitasnya terhadap enzim ini. Namun, tampaknya
masuk akal untuk memprediksi bahwa PAR lain akan
muncul yang diaktifkan oleh enzim lain, bahkan mungkin
protease non-serin. Lebih jauh lagi, studi hubungan
struktur-afinitas ligan oleh Hollenberg dan rekannya
semakin menunjukkan keberadaan subtipe PAR yang saat
ini dikloning10.
Endothelia dan epithelia
Meskipun ada perbedaan yang jelas antara struktur dan
fungsi endotelia dan epitel, kedua tipe sel dicirikan oleh
kemampuan mereka untuk menanggapi potensi tantangan
inflamasi akut dan untuk memodifikasi sifat dan perilaku
mereka dalam jangka panjang. Untuk memanifestasikan
spektrum tanggapan terkoordinasi, kedua jenis sel
menggunakan mediator autokrin dan parakrin. Mediator
ini tidak hanya merupakan sinyal penting bagi sel-sel
penghalang itu sendiri, tetapi juga ke jaringan yang
mereka lindungi dan karenanya merupakan pemain vital
dalam menjaga keseimbangan antara reaksi pertahanan
akut dan keadaan penyakit kronis.
Aktivasi seluler akut
Menanggapi aktivasi seluler akut, sel epitel di seluruh
tubuh melepaskan mediator seperti nitric oxide (NO) dan
prostanoids, yang dihasilkan oleh enzim yang
diekspresikan secara konstitutif. Pentingnya potensi
mediator akting akut ini tidak diakui secara luas, mungkin
karena mereka tidak selalu memiliki efek yang jelas pada
otot polos dalam persiapan dan karena itu sulit dideteksi
dengan teknik bioassay sederhana. Namun, mediator
seperti NO dan prostaglandin (PG) yang dilepaskan secara
akut oleh sel epitel memiliki efek penting dan umumnya
bermanfaat pada sel-sel inflamasi dan proses dalam
mukosa organ berlapis epitel (Gambar 2) 11,12. Penelitian
di bidang vaskuler didominasi oleh konsep bahwa
mediator yang diturunkan dari endothelium terutama
terlibat dalam regulasi tonus pembuluh darah sebagai
akibat dari efeknya yang lebih jelas pada otot polos.
Namun, mediator yang diturunkan dari endotelium seperti
NO dan prostasiklin memiliki peran tambahan dalam
pencegahan agregasi platelet dan penghambatan infiltrasi
dan aktivasi sel imun13. Oleh karena itu, aktivasi akut
epitel dan endoteli memunculkan produksi faktor aksi
pendek yang menentang inisiasi respons inflamasi dan,
karenanya, sangat melindungi organ yang bersangkutan.
Aktivasi seluler kronis
Berbeda dengan aktivasi akut sel epitel dan endotel,
aktivasi kronis sel-sel ini menginduksi respon yang
berbeda yang melibatkan pelepasan mediator seluler yang
disintesis baik de novo dari jalur konstitutif, seperti
banyak anggota keluarga sitokin, atau dengan diinduksi.
mediator tidak hanya memiliki efek dalam epitel dan endotelia,
seperti perubahan fenotip seluler, tetapi juga melibatkan
aktivasi sistem kekebalan tubuh dan perekrutan leukosit dari
sirkulasi. Dengan demikian, sebagian besar peristiwa ini
dianggap pro-inflamasi karena perubahan perilaku sel normal
dan adanya leukosit dalam jaringan biasanya diamati pada
keadaan inflamasi kronis. Namun, dalam homeostasis sehari-
hari yang normal, banyak dari mediator ini memainkan peran
yang menguntungkan - hanya ketika stimulus pro-inflamasi
bertahan bahwa peradangan kronis terjadi.
Pembuluh darah
Trombin memberikan efek jangka pendek dan jangka panjang
pada sel endotelial. Secara akut, trombin menginduksi
pelepasan vaso- dilator dan antikoagulan, konsisten dengan
peran normal endotelium dalam mencegah agregasi trombosit
dan mengatur aliran darah. Dalam jangka panjang, trombin
mengubah fenotip sel endotel menjadi pro-koagulan dengan
menginduksi pelepasan faktor von Willebrand dan ekspresi
faktor jaringan14. Proses ini mungkin memperkuat gumpalan
yang diperlukan, seperti yang awalnya menyatukan luka.
Keterlibatan PAR dalam efek ini perlahan-lahan menjadi jelas.
Pada sebagian besar hewan yang diselidiki sejauh ini, efek
vasodilator dari trombin ditiru oleh peptida pengaktif PAR1,
dan respon trombin dihapuskan oleh desensitisasi
sebelumnya dengan peptida2. Namun, situasinya kurang jelas
pada jaringan manusia. Meskipun sel-sel manusia yang
dikultur dari endotel vena umbilikalis (HUVECs)
mengekspresikan fungsional PAR1 (Pustaka 2), ini tampaknya
bukan kasus di seluruh jaringan yang baru diperoleh15.
Selanjutnya, aksi vasodilator dari trombin tampaknya
dimediasi oleh reseptor atipikal di beberapa arteri manusia16;
reseptor ini diaktivasi oleh trypsin dan trombin, dan oleh ligan
PAR1 (promiscuous) SFLLRN-NH2,
tetapi tidak dengan ligan untuk PAR lainnya. Perbedaan
serupa adalah
jelas ketika efek kronis trombin pada sel endotel manusia
dipertimbangkan. Dengan demikian, walaupun ligan PAR1
meniru efek trombin dalam menyebabkan kontraksi sel
endotel, tidak seperti trombin, ligan ini merupakan agonis
reseptor yang buruk dalam migrasi sel dan tes proliferasi18.
Contoh-contoh ini hanya mulai menyoroti masalah saat ini
yang terkait dengan upaya untuk mendefinisikan PAR
menggunakan ligan peptida yang memiliki karakteristik
kurang baik. Terlepas dari perbedaan antara jaringan kultur
dan asli dalam hal ekspresi reseptor, faktor-faktor lain yang
memerlukan pertimbangan hati-hati termasuk keberadaan
dan aktivitas inhibitor enzim endogen (misalnya serpins) dan
peptidase dalam jaringan yang berbeda, selektivitas ligan dan
kemungkinan adanya reseptor atau varian sambatan yang
belum dijelaskan. Lebih jauh, spesies yang berbeda mungkin
menggunakan PAR yang berbeda untuk fungsi fisiologis yang
berbeda. Sebagai contoh, PAR3 adalah PAR platelet utama
pada tikus, sedangkan PAR1 memenuhi tugas ini pada
manusia19.
Peran untuk PAR2 pada sel endotel kurang jelas karena
identitas enzim pengaktif tidak jelas dalam lingkungan
vaskular. Dua kandidat yang sah, hingga saat ini, adalah
faktor koagulasi Xa (Ref. 20) dan tryptase sel mast2. Namun,
trypsin, yang merupakan aktivator PAR2 paling kuat yang
belum dijelaskan, juga merupakan kandidat karena sel-sel
endotel memiliki
been shown to express trypsin in culture and in patients
with cancer or disseminating intravascular coagulation 21.
Despite the current interest in mast cell tryptase as a
candidate, it is probable that factor Xa plays an
important role in the vas- culature. Being a large
tetrameric molecule, mast cell tryptase would not be
expected to easily traverse the wall of a blood vessel and
thus would be unlikely to reach the endothelium; this
argument is supported by the late appearance of mast cell
tryptase in plasma compared with histamine during
anaphyl- actic reactions22. In addition, although the effects of
activation of PAR2 in vascular preparations from animals
are relatively clear, their role in human blood vessels
is less certain. It
appears that the PAR2-activating peptides SLIGKV-NH 2
and SLIGRL-NH2 have no vasodilator activity in
human large and small coronary and pulmonary arteries
(Ref. 16 and
J.R. Hamilton and T.M. Cocks, unpublished
observations),
whereas powerful PAR2-mediated vasodilatation has
been demonstrated in animal preparations both in
vitro2,23 and in vivo24,25. To date, there is only one study
that suggests that PAR2 might activate human
endothelial cells acutely in whole tissue preparations15.
However, PAR2 has been localized by
immunohistochemistry and in situ hybridization to the endo-
thelium and media of a variety of human arteries26,27
and endothelial PAR2 mRNA and protein have been
detected in human coronary arteries (J.R. Hamilton and
T.M. Cocks, unpublished observations). In addition,
preliminary studies demonstrate that incubation of these
arteries with inter- leukin 1a upregulates endothelium-
dependent relaxant responses to SLIGKV-NH2. These
observations suggest that
PAR2 are expressed by human vascular endothelial cells
but
under normal conditions they cannot be activated suffi-
ciently to cause vasorelaxation. Explanations for this appar-
ent paradox include expression of too few receptors or
fail- ure of expressed receptors to couple to G proteins
adequately to initiate release of endothelium-derived
relaxing factors. On a longer time scale, activation of
PAR2-like receptors has been shown to induce vascular
smooth muscle prolifer- ation28 and phenotypic changes
in endothelial cells29,30, which suggests that PAR2 might
only regulate chronic vas- cular responses in humans. The
expression of trypsin by pro- liferating endothelial cells in
culture and in tumours con- taining growing blood
vessels21 might be highly significant in this regard.
Tethered Enzyme-specific
ligand cleavage site
Hirudin-like sequence
alignment site NH3+
(PAR1 and
PAR3 only)
Exogenous tethered
ligand sequence
Extracellular
TM1 TM7
Intracellular
G-protein
binding
COO
trends in Pharmacological Sciences
Fig. 1. A schematic representation of the mechanism of activation of
protease- activated receptors (PARs) is shown. The ‘tethered ligand’
sequence (blue box), which is exposed following enzyme-specific
cleavage, binds to a site on the receptor (green box), probably on the
second extracellular loop, to initiate G-protein-meditated cell signalling.
Synthetic peptide mimetics of PARs teth- ered ligand sequences can also
activate PARs by binding to the receptor (green box) without enzymatic
cleavage of the receptor.
Epithelia
Although PAR1 does not appear to be functionally
expressed in gastrointestinal and pancreatic epithelia31,32,
both thrombin and PAR1-activating peptides act on
airway epithelial PAR1 to initiate PGE2-dependent
bronchoprotection33. Expression
of PAR1 by epithelial cells might have important functional
consequences. Thrombin can only access epithelia following
damaging insults to these usually tight defensive barriers.
Such insults do not have to be physical because Persson
and col- leagues have demonstrated that plasma exudes
through the epithelial barrier during allergic challenges
that promote plasma extravasation in the submucosa 34. In
addition, when the epithelium is physically disturbed
similar exudative pro- cesses result in a plasma-derived
gel that acts to protect the mucosa while surrounding
epithelial cells grow over and cover the wound34. The
presence of PAR1 on epithelial cells suggests that these
receptors might be involved in both damp- ening
inflammatory processes via activation of PGE2-mediated
cytoprotection and the induction of appropriate longer
term

Table 1. Characteristics of the four cloned protease-activated receptors (PARs)

Property PAR1 PAR2 PAR3 PAR4

Selective enzyme activator Thrombin Trypsin Thrombin Thrombin and trypsin

Chromosome (human) 5q13 5q13 5q13 19p12
Other enzyme activators Trypsin, mast cell Mast cell tryptase
tryptase
Cleavage site (human) Arg41–Ser42 Arg34–Ser35 Lys38–Thr39 Arg47–Gly48
Hirudin-like alignment site Yes No Yes No
TiPS – March 2000 (Vol. 21)
Tethered ligand sequences SFLLRN (h), TFRIFD (x), SLIGRL (m, r), SLIGKV (h) TFRGAP (h), SFNGCP (m) GYPGKF (m), GYPGQV (h)
SFFLRN (m, r)
Activity of synthetic tethered Active Active Inactive Active
ligand sequences
G-protein coupling Gaq/11, Gi, Gao, Ga12, Ga13 Gaq, Gao Unknown Unknown

Abbreviations: h, human; m, mouse; r, rat; x, Xenopus.

106 TiPS – March 2000 (Vol.

 Agonis
t
obvious solution to the apparent paradox of PAR2
(3) Adhesion expression outside the duodenum and pancreas is an
(4) Fluid secretion
(1) Mucus (2) Ciliary equally widespread distribution of trypsin. In addition,
secretion beating although intuition suggests that potentially destructive
enzymes should be confined to sites of digestion and
aberrant cancer cells2, trypsin mRNA and protein are in
fact expressed by a variety of epithelial cell types.
Thus, both PAR2 and trypsin have been localized in the
epithelia of the skin, kidney, lung, intestine and
liver26,33,35. Similarly, PAR2 and trypsin are both
expressed by leuko- cytes2,35. Furthermore, disparate
Epithelium
epithelial cell types produce other pancreatic enzymes such
PGE2
as chymotrypsin36, as well as factor Xa (Ref. 37) and novel
NO
tryptic enzymes38. Hence, many candidate epithelial
PAR2-activating enzymes, other than mast cell tryptase,
Blood
(5) vessel
are emerging. Indeed, epithelial PAR2 might sense
luminal tryptic enzymes (Fig. 3)33 and initiate
cytoprotective pathways that include inhibition of the
sub- luminal release of mast cell tryptase (Fig. 2). In this
model,
Inhibition of tryptase might act only chronically at PAR2 and only
mast cell during prolonged inflammatory reactions.
activation
(6) Vasodilatation Therapeutic horizons
PARs might be targets for novel drugs in many clinical
Inhibition of Leukocyte
settings that involve inflammation. Generally, however,
leukocyte
only pro- longed inflammatory responses attract the
invasion
trends in Pharmacological attention of drug companies that strive to inhibit these
Sciences reactions with receptor
Fig. 2. A schematic representation of the acute cytoprotective effects of prostaglandin E (PGE )
and antagonists. Less attention is paid to the development of
recep-
2 2 tor agonist drugs to activate constitutive, anti-inflammatory
nitric oxide (NO) is shown. Endogenously produced PGE2 and NO (principally of epithelial origin) there have been no studies on the chronic
exert actions both on the epithelium itself and on surrounding tissues 11,12. Epithelial cells respond
to PGE2 and NO by increasing: (1) mucus secretion; (2) frequency of ciliary beating; (3) cell–cell effects of trypsin or PAR2 activation on
adhesion; and epithelial cells, although many studies
(4)net ion transport to the mucosa (encouraging water transport to the lumen). These effects act
in concert to protect the epithelial barrier. PGE2 and NO also inhibit local immune cells (5) and suggest that mast cell tryptase can exert
increase local blood flow (6). Thus, the combined effects of epithelium-derived mediators long-term effects on these and other
orchestrate an acute defensive response to epithelial challenge, limiting access of lumenal cells2.
pathogens and inhibiting more aggressive inflammatory reactions.
Unlike endogenous activators of
endothelial PAR2, those of epithelial PAR2
effects on epithelial cells during restitutive processes such are less difficult to identify. Although inter-
as adaptive phenotypic changes and cellular proliferation. est grows in mast cell tryptase as a PAR2
PAR3 and PAR4 might be similarly involved, although, activator2, a more
to date, no functional studies have been conducted that
demonstrate the presence of these receptors on epithelial
cells. PAR2 is expressed by epithelial cells throughout
the body26. The high levels of PAR2 expression on
epithelia in the intestine and pancreas are most
probably related to the high levels of pan- creatic trypsin
to which these organs are exposed. Thus, acti- vation of
intestinal PAR2 or a PAR2-like receptor by trypsin or
PAR2-activating peptides initiates PGE 2 production by
epithelial cells and subsequent cytoprotective mechanisms
such as promotion of fluid secretion2,31. PAR2 are also
abun- dantly expressed on airway epithelia26,33 and, as
in the gut, their activation by trypsin or PAR2-activating
peptides ini- tiates similar PGE2-dependent
cytoprotection33. Thus, the acute effect of tryptic enzymes
on epithelial cells appears to be mainly protective. To date,
responses that have to be overcome for inflammation to persist. This
second approach might be usefully exploited in the field of PARs
because PAR-activating peptides or small-molecule mimetics could
switch on anti-inflammatory mechanisms without the side-effects that
endogenous enzymes might cause via other mechanisms. The power of
activating endogenous anti-inflammatory pathways should not be
ignored. For ex- ample, antithrombin III activates endothelial
cytoprotective pathways that not only prevent inflammatory processes but
can also reverse them39. Furthermore, in ischaemia–reperfusion injury
the endothelium is in an activated, pro-inflammatory state and yet
antithrombin III binding to the endothelium generates sufficient
quantities of anti-inflammatory paracrine mediators, such as
prostacyclin, to reverse the ongoing in- flammatory response. It is
worth recalling that current thera- pies for inflammatory diseases such
as asthma, Crohn’s disease and arthritis are currently treated with
receptor agonists, notably 13-adrenoceptor (e.g. in asthma) and steroid
receptor (e.g. in most inflammatory reactions, including persistent
asthma) agonists. By activating endogenous anti-inflamma- tory
cascades, pharmacologists might develop unique treat- ments for
chronic inflammatory diseases.

The vasculature
The most obvious way to harness PARs therapeutically is in the area
of blood coagulation in which receptor agonists and antagonists
could be used to control platelet activation. Simi- larly, if the pro-
inflammatory actions of thrombin can be ascribed to a specific PAR,
antagonists for this receptor might have therapeutic value. The
challenge that lies ahead is to define which actions of thrombin are,
in fact, mediated by
PARs (rather than other thrombin receptors40) and
Trypsin
which PAR or PAR subtype is responsible. Because PAR2 (regulated acute
does not appear to be directly involved in haemostasis, mediator)
PAR2 agonists hold great promise as agents that could
be used to activate endogenous protective pathways
without side-effects on blood coagulation. Neither
PAR1 nor PAR3 expression on endothelial cells is
affected by pro-inflammatory mediators such as tumour
PAR2
necrosis factor a or interleukin 1a, suggesting non-
adaptive regulation of expression41. By contrast, similar PAR2
mediators increase PAR2 expression2 and the vasodilator
function of PAR2 on endothelial cells in vitro and
PAR2 expression appears to be elevated in
endotoxaemic rats42. Although it has been suggested that
such observations impli- cate PAR2 in the progression PAR2
Disrupted
of inflammation2, the exact opposite might equally
epithelium
apply. That is, PAR2 upregulation confers increased
protection. For example, an increased potential for ?
thrombosis is frequently observed in humans and in animal
models of sepsis induced by lipopolysaccharide (LPS)43. Tryptase
Therefore, upregulation of both endothelial PAR2 (the
sensor) and coagulation (possibly the activator) in response
to such pro-inflammatory stimuli such as LPS suggests that (unregulate
d
PAR2 might play an important role in limiting clotting chronic
and preventing disseminating intravascular coagulation. mediator)
This sug- gestion is supported by the observation that trypsin
Chronic Acute
is expressed by endothelial cells in arteries from patients inflammation cytoprotection
suffering from this pathology21. Similarly, hypertension is
frequently associated
with decreased receptor-mediated endothelium-dependent identified. However, not all cellular effects of enzymes are, or will be,
relaxation to most vasodilator agents44 and subsequently mediated by PARs. Many of these effects might be mediated by biologi-
with an increased potential for thrombosis45. By cally active enzyme products or by cell-surface receptors not
contrast, PAR2- mediated endothelium-dependent
responses are preserved in hypertensive rats46. This
correlation between increased thrombotic potential and
PAR2 expression in endothelial cells again could indicate
protection by PAR2, rather than a role for PAR2 in
hypertension. Such a conclusion is supported by the
above suggestion that PAR2 present on human endo-
thelial cells only couples to dilator pathways if
stimulated by inflammatory cytokines. Hence, in both
sepsis and hyper- tension endogenous anti-coagulant,
vasodilator mechanisms, or both could be activated
selectively to provide therapeutic positive feedback.

Epithelia
Epithelial PARs might provide the most attractive targets
for novel drug development because many epithelia can
be tar- geted without the need for systemic drug
administration. Inhalation of a PAR2 tethered ligand
sequence has recently been shown to perform this task
well in rats33. Therefore, novel PAR-agonist-based
recruitment of existing anti-inflammatory mechanisms
might have exciting applications in diseases of other
mucosal surfaces.

Concluding remarks
Biochemists who have explored the cellular effects of
proteases in detail must be delighted that targets for
some protease- mediated actions can be tentatively
 trends in
Pharmacol
ogical
Sciences

Fig. 3. A schematic two-compartment model that describes the

functionality of apically restricted epithelial protease-activated
receptor PAR2 in the airways is shown. Tight junctions between cells
(mauve boxes) maintain a barrier to diffusion of macromolecules. Hence,
any epithelium-derived trypsin (green circles) is likely to be contained at
the apical surfaces of the epithelial cells. By contrast, mast cell tryptase
(red circles) is likely to be restricted in the submucosa (or below the
tight junction barrier if released by mast cells within the epithelium) and
is therefore unlikely to activate apical PAR2 if the epithelium is intact.
Because trypsin is sensitive to endogenous inhibitors produced in the lung
(e.g. a1-antitrypsin), it can act only as an acute mediator, whereas mast cell
tryptase, which has no known endogenous inhibitors and is thus
unregulated, might act only as a chronic mediator. The dashed lines
indicate that mast cell tryptase might act inappropriately on either
lumenal PAR2 if the epithelium is damaged by, for example, eosinophil-
derived major basic protein during allergic reactions, or possibly on basal
PAR2 if they are present in this location.

related to PARs. Furthermore, the use

of synthetic peptide receptor agonists
points increasingly towards the
existence of subtypes of the cloned
PARs, and proteases other than
thrombin, trypsin, factor Xa and mast
cell tryptase might exist for which
PARs act as sensors. These ‘new’
PARs not only need to be cloned and
expressed, but additional functional
data – particularly from human tissues –
is equally important to allow the
continued exploration of the potential
of PARs as therapeutic targets.

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