Bioactive polysaccharides from
medicinal plants used in
woundhealing and against
inflammation
Berit Smestad Paulsen
Department of Pharmacognosy
School of Pharmacy
Oslo, Norway
MALI
http://www.xs4all.nl/~ilja/mali/mali-map-htm
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Malian population:
Population: ca 12 mill.


Area

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
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47.2% : 0-14 years
5,5% : > 60 years
Fertility rate: 6.7 children born/woman
Child mortality: 12%
Rate of alphabetization:
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1 241 248 km2
Expected living age: 47 years
Age repartition:

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Ethnic diversity: Sénoufo, Bambara, Sarakolé, Milinké, Peulh,
Dogon, Bobo, Sonrhaï, Tamacheck……..
Men
– 31%
Women – 15%
Appr. 75% in rural areas
Photo: C.S. Nergård
Curative urban based health
service,
 limited curative or preventive
health service in the rural
areas
 59% of the population have
healthservice within 15 km

Medical doctors
1:16,000 inhabitant
(Norway: 1:272)

Local clinics:


Department of
Traditional
Medicine
(DMT)
difficult to keep the
personnel
Funding not available for long
time treatment
Traditional medicine is composed of:
-Secret traditional medicine
TRADITIONAL HEALERS
-Methods of diagnosis
1:500 inhabitants
-Methods of treatment
80% use traditional medicine
Ethnopharmacology

The observation, identification, description and
experimental investigation of the ingredients and
the effects of such indigeneous drugs is a truly
interdisiplinary field of research which is very
important in the study of traditional medicine
 The interdisiplinary scientific exploration of
biologically active agents traditionally employed
or observed by man.
Traditional preparation and adminiatration
 Collection

of the plant part
Use either fresh or dry
 Preparation


Dry powder
Infusion or macerate
 Normally
extract
wash the body or drink the
Preparation of the polysaccharides

Ethanol extraction
 Water extraction 50oC




Dialysis, freezedrying
Anion exchange chromatography
gelfiltration
Water extraction 100oC



Dialysis, freezedrying
Anion exchange chromatography
gelfiltration
Structure elucidation
 Methanolysis,

Carbohydrate composition
 Methylation

TMS GC
studies GC-MS
Linkage determination
 NMR
anomeric configuration, sequence
 Mw - gelfiltration
 Enzymatic degradation

Gives polygalacturonans, hairy regions,
oligosaccharide sidechains etc.
Immunomodulation associated with
activation of the complement system:






Thymus dependant antibody response
Regulation of specific cyclic antibody production
Regulation of IgM-IgG switch
Modulation of T and B cell proliferation
Induction of supressor or helper T-cells
Modulation of monokine or lympokine release
Structures of pectic type
polysaccharides
 Arabinogalactan

1,4,gal
 Arabinogalactan

type I
type II
1,3,6 gal
 Rhamnogalacturonan

Hairy region, alternating rha / galA backbone
 Rhamnogalacturonan

I
II
Macro-oligosaccharide, 20 units, complex
Proposed structure of pectins, Perez et al 2003
Hairy region of
rhamnogalacturonan I
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

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
g2Rha1g4GalA1g2Rha1g4GalA1g2Rha1g4GalA1g2Rha1g4GalA1g
4
4
4
h
h
h
1
1
1
Araf
Gal
Gal
5
6
6
h
h
h
1
1
1
Araf1g3Araf
Araf1g3Gal
Gal
5
6
6
h
h
h
1
1
1
Araf
Gal
Araf1g3Gal
5
6
6
h
h
h
1
1
1
Araf1g3Araf
Araf1g3Gal
Gal
5
6
h
h
1
1
Araf
Araf
Are polysaccharides transported
over membranes?
 Peyer´s
patches
 Yamada et al. produced antibodies against
bupleuran
 Detected polymers with binding sites for
the antibodies in liver
Bupleuran epitopes
Medicinal plants used for
woundhealing in Mali
 Baccharoides
adoensis var. kotschyana
(Sch.Bip.ex.Walp.) M.A.Isawumi, G.ElGhazaly & B.Nordenstam (Vernonia
kotschyana (Sch.Bip.ex.Walp.))
 Cochlospermum tinctorium Perr.
 Glinus oppositifolius (L.) A. DC.
 Biophytum petersianum Klotzch
Baccharoides adoensis var.
kotschyana (Vernonia
kotschyana)
Powdered roots are
recognised by the
government in Mali as
a natural drug for the
treatment of
gastroduodenal
ulcers.
GASTROCEDAL
Characteristics of A.2
Acacia gum (std.)
A,2,2
Vk100
A1
A2
A.2.1
Gelfiltration of A.2 from V.kotschyana. Effect
on the Yariv reagent, separation after enzymic
degradation of A.2.2
A2.1
A2.2
Effect of A.2.1 after enzymatic degradation
Effect of A.2.2 after enzymic
degradation
Vk100A2a
10 g/mL
1
Vk100A2b
1,5
0,5
0,0
Vk100A2a
1
Vk100A2b
B-cells
6,0
*
0,0
Vk100A2a
1
Vk100A2b
BR2IIc 100 g/mL
*
ConA 5 g/mL
*
Anti-IgM F(ab)2 10 g/mL
*
10 g/mL
1,0
1,5
8,0
100 g/mL
1,0
(B)
10 g/mL
*
Spleen-cells
100 g/mL
Acid phosphatae
Control
*
Stimulation index (SI)
2,0
BR2IIc 100 g/mL
*
100 g/mL
3,0
2,5
10 g/mL
3,5
100 g/mL
(A)
10 g/mL
Alamar blue
Control
*
OD 405
Spleen-cells
BR2IIc 100 g/mL
4,0
ConA 5 g/mL
4,5
Anti-IgM F(ab)2 10 g/mL
5,0
100 g/mL
2,0
10 g/mL
2,5
100 g/mL
0,0
Control
0,5
Control
Stimulation index (SI)
Mitogenic activity of fractions
from V. kotchyana
Alamar blue
*
(C)
*
7,0
5,0
*
4,0
3,0
*
2,0
1,0
Macrophages
A
Anti-CD14
B
Donor 1
Donor 4
9000
8000
7000
8000
6000
5000
4000
3000
6000
2000
1000
0
2000
7000
5000
4000
3000
1000
0
Vk100A2a
7
Vk100A2b
11
LPS
12
250 ng/ml
10
25 ng/ml
9
2.5 ng/ml
8
500 g/ml
7
250 g/ml
100 g/ml
6
13
10 g/ml inulin
Vk100A2a
5
10 g/ml
4
500 g/ml
250 g/ml
3
100 g/ml
10 g/ml
2
9
Vk100A2b
10
9
8
7
6
5
4
3
2
1
0
1
8
10
11
CXCL12 100 ng/ml
6
10 g/ml
5
100 g/ml
4
1 g/ml
3
Medium
2
100 g/ml
1
1 g/ml
11
10 g/ml
10
Medium
9
Vk100A2b
Medium
Nitrite (g/ml)
C
8
CXCL12 100 ng/ml
Vk100A2a
7
100 g/ml
6
1 g/ml
5
10 g/ml
4
Medium
3
100 g/ml
1 g/ml
2
10 g/ml
1
Medium
# of migrated cells
9000
Stimulation of
macrophages
with Vk100A2
fractions,
chemotaxis in
response of
diff. conc. in 2
diff. patients
Yield (%, w/w), total carbohydrate content (%, w/w) and
monosaccharide composition (mol.%) of the fractions Vk100A2a and
Vk100A2b obtained after size exclusion chromatography on Sephacryl
S-400 of Vk100A2.
Polysaccharide fraction
Yield (%, w/w) a
Total carbohydrate (%, w/w)
Monosaccharide composition (mol.%)
Arabinose
Rhamnose
Galactose
Xylose
Mannose
Glucose
Fucose
Glucuronic acid
Galacturonic acid
Reaction with Yariv-reagent
Protein content (%, w/w) b
a based on dried and pulverized root
b based on the Lowry method
Vk100A2
a
b
0.05
0.34
86
68
31.3
11.0
24.4
0.7
0.3
5.7
trace
26.4
+
< 0.5
3.9
4.9
4.9
trace
trace
1.8
trace
84.6
< 0.3
Vo
1,6
A
1,4
Absorbance
1,2
Vk2a-HR
1
0,8
0,6
0,4
Vi
0,2
0
11
15
19
23
27
31
35
39
43
47
51
55
59
63
67
71
75
Vo
0,7
79
83
B
0,6
0,5
AF-HR
Absorbance
0,4
0,3
0,2
Vi
0,1
0
11
15
19
23
27
31
35
39
43
47
51
55
59
63
67
71
75
79
83
-0,1
C
AF-GN-3
0,16
AF-GN-1
0,14
Absorbance
0,12
AF-GN-5
AF-GN-4
AF-GN-6
AF-GN-7
AF-GN-2
0,1
AF-GN-8
0,08
0,06
0,04
0,02
0
11
15
19
23
27
31
35
39
43
47
51
Fraction no.
55
59
63
67
71
75
79
83
Degradation of VKA2 by
a, polygalacturonase
followed by
b. arabinofuranosidase
followed by
c. galactanase
Effect of the different fractions on the
complement system
100,0
Vk2a
% Inhibition of hemolysis
80,0
Vk2a-HR
AF-HR
60,0
AF-GN-1
AF-GN-2
AF-GN-3
40,0
AF-GN-4
AF-GN-5
20,0
0,0
0
-20,0
5
10
15
20
25
Sample concentration (g/mL)
30
35
B cells
10,0
A
9,0
*
Stimulation index (SI)
8,0
*
Mitogenic activity
on the original
fraction and the
degradation
products
*
7,0
6,0
*
5,0
*
4,0
*
3,0
*
2,0
AF-GN-2
AF-GN-1
AF-HRGN-mix
1
AF-HR
Vk2a-HR
Vk2a
BR2IIc
0,0
Control
1,0
Sample concentration (100g/mL)
Spleen cells
B
2,5
*
2,0
*
*
*
1,0
*
Sample concentration (100g/mL)
AF-GN-2
AF-GN-1
AF-HRGN-mix
1
AF-HR
Vk2a-HR
Vk2a
0,0
BR2IIc
0,5
Control
OD (405 nm)
*
1,5
Conclusion
 Bioactive
parts on the VK1002a
polysaccharide are situated both in the
inner and in the external part of the pectic
arabinogalactan
Cochlospermum tinctorium, root

The root is
traditionally used
agains gastric ulcer,
often together with
the powder from the
root of V. kotchyana
Cochlospermum tinctorium
activity on whole spleen cells
Cochlospermum tinctorium
activity on B - cells
Glinus oppositifolius (L.) A. DC. (Aizoaceae)
Used
in
traditional
medicine for the treatment
of
joint
pains,
inflammations,
fever,
malaria, skin disorders and
wounds.
 The immune system an
important factor in the
wound healing process.
 Plant
polysaccharides
reported
to
have
immunomodulating
activities.

Glinus oppositifolius (L.) A. DC. (Aizoaceae)

50°C water extract separated into one neutral
(GON) and two acidic fractions (GOA1 and
GAO2).
M on osa c ch aride
GO
Arabi n os e
9
Rh am n os e
7 ,2
F uc os e
1 ,3
Xy lose
1 ,1
M an no s e
1 ,7
Galac t os e
14 ,9
Galac tur on i c aci d
58 ,9
Gluco s e
5
4 -O-M e -Glucu ro n ic
1 ,1
ac id
Mole cu la r we igh ts
( De ter m ine d us in g SEC/ MALLS )
GON
32 ,2
0 ,1
0 ,5
2 ,3
5 ,1
34 ,5
25 ,4
-
GOA1
23
4
3 ,4
4 ,5
44 ,6
13 ,4
3 ,6
3 ,5
7 0kD a
GOA2
4 ,4
9 ,1
1 ,1
0 ,4
0 ,6
9 ,2
71 ,5
3 ,2
0 ,5
39k Da
The monosaccharide compositions of the crude water extract
GO and the different fractions obtained after ion exchange
chromatography.
Complement fixating activity I
The complement fixating activities determined in vitro.
GOA1 shows an ICH50 value at 40 g/ml (the lowest concentration of
sample needed to give 50% inhibition of lysis of antibody-sensitized
SRBS). Plantago major L. was used as a positive control.
% inhibition of hemolysis


100
90
80
70
60
50
40
30
20
10
0
GOA1
GOA2
GON
PMII
0
50
100
150 200 250 300 350
400 450 500
Concentration ug/m l
Complement activation associated with several immune reactions; activation of macrophages
and lymphocytes, immunopotenation and regulation of cyclic antibody production, antiinflammatory effects.
Linkage analysis
Type of
linkage
T-Araf
Ara 1,3
Ara 1,5
Ara 1,3,5
Ara 1,2,5
GOA1
Original Hydrolysed
5,4
0,03
1,6
0,06
3,9
0,05
1,1
0,03
0,2
0,03
T-Rha
Rha 1,2
Rha 1,2,4
1
0,4
0,7
0,3
0,5
0,8
T-Xyl
Xyl 1,4
Xyl 1,2,4
0,9
0,6
0,2
0,2
0,4
0,06
Man 1,2
2,3
1,5
T-Galf
T-Galp
Gal 1,4
Gal 1,3
Gal 1,6
Gal 1,3,4
Gal 1,4,6
Gal 1,3,6
Gal 1,3,4,6
0,2
2
2,8
3,6
4,2
0,3
0,1
8,6
1,6
0,09
1,5
1,8
1,9
4,8
0,1
0,05
2,8
0,05
T-Glc
Glc 1,4
Glc 1,4,6
0,4
1,1
0,4
0,2
0,6
0,1
T-GalA
GalA 1,4
1,3
5,7
2,0
4,7
T-4-O-Me-GlcA
1,3
4-O-Me-GlcA 1,4 0,5
1,2
0,3
The main linkages in GOA1:
GalA 14
Rha 12, 12,4
Gal 1 4, 1 3,4
Gal 1 3, 1 6, 1 3,6, 1 3,4,6
Araf T, 1 3, 1 5, 1 3,5
The migration of different
leukocyte populations in
response to GOA1.
# of migrated cells
Chemotaxis of macrophages,
B-cells, and T-cells.
5
4
3
2
1
0
Medium
100 ug
SB1
10 ug
SB1
1 ug SB1
Medium
100 ug
SB2
10 ug
SB2
1 ug SB2
SDF-1
Macrophages
T-cells
IP
-1
0
l
ng
/m
ug
/m
l
1
10
0
O
A2
O
A2
G
l
ug
/m
l
G
ug
/m
10
G
O
A2
ed
iu
m
10
0
M
G
O
A2
l
ng
/m
ug
/m
l
G
O
A1
1
10
0
ug
/m
l
l
0
O
A1
GOA2 GOA2 GOA2 1 GOA2
100 10 ug/ml ug/ml
100
ug/ml
ng/ml
G
Medium
ug
/m
GOA1 GOA1 GOA1 1 GOA1
100 10 ug/ml ug/ml
100
ug/ml
ng/ml
10000
G
Medium
20000
10
0
30000
O
A1
5
40000
G
10
50000
ed
iu
m
15
60000
M
20
10
0
25
O
A1
# of migrated cells
# of migrated cells
B-cells
Conclusion


Two different pectic polysaccharides isolated from
Glinus oppositifolius (L.) A. DC. (Aizoaceae) with
potent complement fixating activity.
Further structure- and activity studies in progress.
A complement
fixing
polysaccharide
from Biophytum
petersianum
Klotzch, a
medicinal plant
from Mali, West
Africa.
Bp 100, Fr. III, and degradation fraction of Fr. III with
the polygalacturonase, Fr.III 1 is the hairy region








Monosaccharide
Arabinose
Rhamnose
Fucose
Xylose
Mannose
Galactose
Galacturonic acid
Fr.III
8,5
7,3
0,8
7,2
0,4
9,0
64,1
1
7,9
22,6
1,1
5,0
2,0
20,0
38,5
2
11,1
12,3
2,3
7.1
0,8
8,5
55,2
3
10,4
7,7
1,9
9,4
1,0
6,4
60,8
Effect of polysaccharide fractions from B.petersianum on the complement
system
120
100
inhibition %
80
Bp100
Bp100 -I
60
Bp100-II
Bp100-III
PMII
40
20
0
0
100
200
300
400
500
600
Linkage analyses of the different monosaccharides
present in the fraction Bp 100 Fr.III and the
fractions after polygalacturonase treatment

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




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Type of link
T-Ara
1->5 Ara
T-Xyl
1->2 Xyl
1->4 Xyl
T-Rha
1->2 Rha
1->3 Rha
1->3,4 Rha
1->2,4 Rha
T-Fuc
T-Gal
1->3 Gal
1->6 Gal
1->3,6 Gal
T- GlcA
T- GalA
1->4 GalA
1->3,4 GalA
1->2,4 GalA
B.P Orig
6,2
2,7
l6,4
0
1,2
1,2
3,4
0,8
0,5
2,5
1
3,1
2,2
1,1
3,3
tr
2,5
52,3
8,5
1,1
B.P.I
7
1,1
4,9
0
0,6
2,4
14,2
0,8
0
6,3
1,3
6,8
4,9
3,1
6,2
tr.
2,3
30,2
6,8
1,1
B.P.II
8,5
0
6,1
1,3
3,1
5,2
2,5
2,9
1,8
0,7
2,4
5
3,4
0
0,6
B.P.III
8
0
9,7
0
2,8
2
3
1,6
1,6
0,5
2,1
1,9
3,7
0
1,4
4,1
31,2
17,9
3,3
6,4
30,1
22,2
3
Major features of the regions of BP
 BP

Hairy region, alternating GalA and Rha,
sidechains of 1/3 of Rha
 BP

II MMW
High GalA, 1/3 branched
 BP

I HMW
II LMW
High GalA, 2/3 branched
Conclusions



Activity compared with structural differences indicate that
the hairy region is important for the activity of the native
polymer.
It appears also that the regions of the original polymer
containing sections of galacturonic acids that are highly
branched, also are important for the activity when
comparing those fractions with that of PMII not having
long sections of branched uronic acid chains.
Comparing the activity of Fr.III.1 with that of the native
polymer, it is also obvious that the regions of the
polymers giving rise to the other two fractions have an
inhibitory effect on the activity of the total polymer.
Bioactive polysaccharides from the
stems of Acanthus ebracteatus
Sanya Hokputsa,
Stephen Harding,
Kari Inngjerdingen,
Kornelia Jumel,
Terje Michaelsen,
Thomas Heinze,
Andreas Kochella
and Berit Smestad
Paulsen
Traditional use in Thailand and
other countries in the region

Sea holly, a mangrove plant
 In Malaysia,


In China


seeds against cough and boils
hepatitis, lymphoma and astma
In Thailand



Root and stem for skin diseases and longevity
Crushed leaves on inflamed joints
Decoction of the whole plant against inflammation
Separation on DEAE-Sepharose, 50oExtract
3.5
2.0
concentration of N aC l
A 602
1.5
2.5
2.0
1.0
1.5
A 601
1.0
0.5
0.5
0.0
0.0
0
50
100
150
200
F raction num ber
250
300
NaCl (M)
Absorbance at 490 nm
3.0
Separation on DEAE-Sepharose, 100oExtract
3.5
2.0
concentration of N aC l
A 1002
1.5
2.5
2.0
1.0
A 1001
1.5
1.0
0.5
0.5
0.0
0.0
0
50
100
150
200
Fraction num ber
250
300
NaCl (M)
Absorbance at 490 nm
3.0
Table 1.
Appearance,
yield, protein
content and sugar
composition
(mol%) of
polysaccharide
fractions
obtained after
separation of the
crude extracts on
the DEAESepharose Fastflow column
60C water extract
100C water extract
A601
A602
A1001
A1002
cream,
fluffy
white,
fluffy
white,
fluffy
white,
fluffy
Yieldsa
20.0
50.2
10.1
48.9
Protein content
(w/w)
4.8
6.4
5.9
6.2
Ara
9.1
11.0
13.3
6.1
Rha
0.9
12.3
0.8
7.5
Fuc
trace
0.8
trace
0.3
Xyl
0.6
1.4
1.1
0.8
Man
3.6
1.4
2.3
0.4
Glc
4.3
3.1
9.9
2.2
Gal
55.0
18.7
39.6
11.0
3-O-Me-Gal
26.5
1.3
33.0
1.9
-
50.0
-
69.8
Appearance
Sugar compositionb
GalA
a calculated
as weight % of
applied material
b mol% of total carbohydrate
content
Effect of the fractions in the
complement system
Table 2. Glycosidic linkage composition (mol%) of the polysaccharide fractions obtained
after separation of the crude extracts on the DEAE-Sepharose Fast-flow column
Sugar
Type of
linkage
Fraction
A601
A602
A1001
A1002
Tf
1.8
2.5
2.7
2.3
1,3
1.0
-
0.7
-
1,5
4.9
7.2
6.0
3.8
1,3,5
1.4
1.3
2.1
-
1,2,5
-
-
1.8
-
T
-
3.1
-
-
1,2
-
7.3
0.8
7.5
1,2,4
-
1.9
-
trace
Fuc
T
-
-
trace
0.3
Xyl
T
-
-
1.1
0.8
Man
T
3.6
1.4
2.3
-
Glc
T
trace
trace
0.9
trace
1,4
4.3
3.1
7.9
-
1,6
-
0.4
-
1,4,6
-
-
0.6
-
Tp
7.8
3.5
8.5
1.7
Tf
4.1
-
1.5
-
1,4
55.8
9.8
57.9
5.3
1,6
4.6
1.4
2.3
-
Ara
Rha
Gala
aas
1,4,6
9.5
3.9
4.4
combined amount of galactose and 3-O-Me-galactose
1,2,4
1.4
5.3
0.5
Linkages present

Neutral polymers
 Arabinose, f (minor)



Acidic polymers
 Arabinose, f
T; 1,3; 1,5M; 1,3,5
Galactose, p (major)

T; 1,4M; 1,6; 1,4,6

Incl. 3-O-methyl-Galactose


Rhamnose, p



T; 1,5
T; 1,2M; 1,2,4
Galactose, p

T; 1,4m; 1,4,6

Incl. 3-O-methyl-Galactose
Galacturonic acid, p

T; 1,4M
CH3-O-C
1
H -N M R
NMR of the Neutral Fraction
Acetyl
6
5
4
3
2
1
ppm
13
108,5 – a C1 L-Araf
C -N M R
104,5 – b1-4 gal
105,3 – b1-4 3-O-megal
58,5 – OCH3
21,2 and 174,8 –
acetylgroups
180
160
140
120
100
ppm
80
60
40
20
1
H -N M R
13
6
5
4
3
2
C -N M R
1
ppm
180
160
140
120
100
80
60
ppm
Summary of 13C chemical shifts for A1002
Chemical shift (ppm)
Residues
C-1
C-2+3
C-4
C-5
C-6
a-(14)-GalpA
100,2
69.2-69.7
79.2-79.7
71.5
175.8
6-Me-GalpA
101.0
69.2-69.7
79.2-79.7
71.5
171.7
b-(14)-Galp
104.4
72.5
n.d
n.d
61.8
a-(15)-Araf
108.6
n.d
n.d
n.d
-
a-(12)-Rhap
n.d
n.d
n.d
n.d
17.6
40
20
Gelfiltration of the acidic fraction 1002
1.8
A 1002e
Absorbance at 490 nm
1.6
1.4
A 1002d
1.2
A 1002c
1.0
A 1002b
0.8
0.6
A 1002f
A 1002a
0.4
0.2
0.0
100
200
300
400
E lution volum e (m l)
500
600
Table 4. Yield, weight average molecular weight (Mw) and sugar composition (mol%) o
f A1002 sub- fractions obtained after size exclusion chromatography of A1002
Fraction
A1002a
Yieldb
A1002b
A1002c
A1002d
A1002e
A1002f
PM IIa
8.6
8.6
24.0
16.0
31.4
3.4
150030
27620
586
423
324
294
Ara
7.3
7.1
5.6
4.3
3.9
4.3
8.8
Rha
5.9
5.7
5.6
5.5
5.5
5.7
4.2
Fuc
0.6
0.3
0.3
0.3
trace
0.3
-
Xyl
0.7
0.6
0.6
0.6
0.5
0.6
-
Man
1.4
-
-
-
-
-
-
Glc
5.1
2.1
1.5
1.6
1.5
2.3
7.3
Gal
16.2
13.1
10.0
8.2
6.9
8.1
8.0
3-O-Me-Gal
13.9
1.8
0.5
trace
trace
trace
-
Total neutral sugars
51.1
30.7
24.1
20.5
18.3
21.3
28.3
49.1
69.4
75.9
79.5
81.8
78.7
71.1
Molecular weight
(KDa)
46-48
Sugar compositionc
GalA (A)
data obtained from Samuelsen et al. 1996
as weight% of applied material
c mol% of total carbohydrate content
a
bcalculated
0 .3
0 .2
0 .1
0 .0
5
10
15
20
25
9 0 º d e te c to r
R I d e te c to r
0 .1 4
Detector response (volts)
Detector response (volts)
0 .1 6
9 0 º d e te c to r
R I d e te c to r
0 .4
0 .1 2
0 .1 0
0 .0 8
0 .0 6
0 .0 4
0 .0 2
0 .0 0
30
5
10
E lu tio n v o lu m e (m l)
20
25
30
9 0 º d e te c to r
R I d e te c to r
1 .4
1 .2
1 .0
0 .8
0 .6
0 .4
0 .2
0 .0
5
10
15
20
25
30
0 .0 9
E lu tio n v o lu m e (m l)
1002a
1002b
1002c
1002d
1002e
1002f
0 .0 8
Detector response (volts)
Detector response (volts)
1 .6
15
E lu tio n v o lu m e (m l)
0 .0 7
0 .0 6
0 .0 5
0 .0 4
0 .0 3
0 .0 2
0 .0 1
0 .0 0
-0 .0 1
10
12
14
16
18
20
22
E lu tio n v o lu m e (m l)
24
26
Inhibition of lysis (%)
100
Effects on the
complement system
80
60
40
20
P M II
A 1001
A 1002
0
0
50
100
150
200
250
C oncentration (  g/m l)
P M II
A 1002c
% Inhibition of lysis
100
A 1002
A 1002d
A 1002a
A 1002e
A 1002b
A 1002f
80
60
40
20
0
0
5
10
15
20
25
C oncentration (  g/m l)
30
35
Conclusion
 High
mol.weight fraction of 1002 highest
activity
 May be one or two types of molecules,




If so:
One basically pectic like
One rich in 3-O-methylgalactose
Or one with all components
 The
effect of the polysaccharides may
explain the traditional use in inflammation
Plantago major
Traditional use in
Scandinavia:
Wound healing
PM II
 De-esterification
and de-acetylation
 Pectinase
 Oxalic
acid hydrolysis
 TFA-hydrolysis
 Pectolyase
 Periodate oxidation
Structure/activity relations

De-esterification and de-acetylation




Hairy regions more active than smooth areas
HMW hairy regions with 1,3,6 linked galactose
highly active
Removal of arabinose on galactose


no change of activity
increase activity
Removal of arabinose on galacturonic acid

reduce activity
PM II activate the complement system
Classical pathway activator
Alterantive pathway activator
not through C1Q (as IgG)
not through the lectin pathway
probably through C3
Variation in activity between different sera, 220 fold
No difference in the level of antibodies against PMII in
the sera
Variation in individual effect
PMII protects against systemic
Streptococcus pneumoniae Infection in Mice
 PMII
protects against the infection when
administered systematically prechallenge
 Protective effect is due to stimulation of
the innate immune system, not the
adaptive.
Conclusions

Polysaccharides from different medicinal plants
from various countries that traditionally are used
as woundhealing agents or against inflammation
have bioactivities that influence the
immmunsystem in different ways in vitro, most
probably also in vivo.
 Possible agents or leads for safe medicines as
no toxic effects yet shown for polysaccharides of
these types.
Co-workers










Cecilie Sogn
Kari Inngjerdingen
Drissa Diallo
Haruki Yamada
Hiroaki Kiyohara
Terje Michaelsen
Sanya Hokputsa
Stephen Harding
Anne Berit Samuelsen
Hilde Barsett
Financial support:
NUFU,PRO 22/2002
Medicinal plants from Mali
COST D28 Therapeutic
polysaccharides