Indian Journal of Experimental Biology
Vol. 39, December 2001, pp. 1308-1310
An experimental study of some indigenous drugs with special reference
to hydraulic permeability
L Upadhyay, A Mehrotra, A K Srivastava• , N P Rai• & K Tripathi*
Department of Mediciile and •Department of Kayachikitsa, Institute of Medical Sciences,
Banaras Hindu University, Varanasi 221005, India
Received 9 August 2000; revised 11 August 2001
The effec t of co mmonly used indigenous drugs for hepatic di sorders i.e. Tinospora cordifolia, (Guduchi/Amrita), Andrographis paniculata (Kalmegha), Picrorhiza kurroa (Kutki), Phyllantnus niruri (Bhoomyamalaki) and Berberis aristata
(Damharidra) was tested on the hydraulic permeability of water in the presence of bile salt through a transport cell model.
The data on hydrauli c permeability were calculated as t (time). JV = Lp x L'.P, where Lp =hydraulic conductivity and L'.P is
the pressure difference. It was observed that the value of controlled hydraulic permeability (0.49 x 10·8 M 3 5" 1 N. 1) decreased in the presence of indigenous drugs and bile salt. The results suggest that these drugs might have the cell membrane
stabilizing property which may lead to prevention of the toxic effect of bile salts in various hepatic disorders.
Biological membrane consisting of lipid and proteins
play a crucial role in almost all cellular phenomena in
the living cells. Due to the complexity of the biological membranes, similar model system are used to provide insights for understanding similar and comparable phenomena in biological membranes. The bilayer
lipid membrane, al so called black lipid membrane
1
(BLM) developed by Tien et al. , is the most widely
experimented model , but due to fragile structure it is
2
very difficult to work with BLM. Srivastava et al. , •3
developed an alternative system consisting of bilayers
of liquid membranes generated from constituent lipids
biomembranes (e.g. lecithin, cholesterol) on a hydrophobic supporting membrane to mimick the action of
biological agents. Thus, if two compartments separated by a hydrophobic supporting membrane are each
filled with a surfactant solution of concentration equal
to its critical micelle concentrates (CMC) a bilayer of
liquid membrane would be formed. The hydrophobic
portions of the surfactant molecules in the liquid
membrane will be preferentially oriented towards the
hydrophobic supporting membrane and the hydrophilic moieties would be drawn towards away from
4
it • The liquid membrane hypothesis by Kesting et
a/. 5 , also showed that when surfactant is added to an
aqueou s phase, the surfactant layer, which forms
spontaneously at the cellulose acetate membrane/saline solution interface, acts as a liquid membrane in series with the supporting membrane. Latter
*Correspondent author: Fax: 0542-3 16068
Srivastava et al. 6 investigated the workability of liquid
membrane bilayers and their role in the mechanism of
action of surface active drugs.
Many indigenous drugs have been used in the
treatment of various hepatic disorders particularly in
viral hepatitis where there is increased concentration
of bile salts and its components in the biliary canaliculi. These drugs are Tinospora cordifolia, (Guduchi/
Amrita), Andrographis paniculata (Kalmegha),
Picrorhiza kurroa (Kutki), Phyllantnus niruri
(Bhoomyamalaki) and Berberis aristata (Daruharidra). The exact mechanism of action of these drugs
have not yet been well defined except few (Phylanthus niruri) where some antiviral property have been
demonstrated. One of the novel explanation of these
drugs are that they may affect the hydraulic permeability of water in the presence of bile salt.
Thus, keeping above facts in view the present study
was planned to investigate the effect of these drugs,
separately and .in combination, on the hydraulic permeability in the presence of lecithin cholesterol and
bile salt through transport cell model.
The hydraulic permeability of indigenous drugs
were studied on transport cell model as described by
Srivastava et al. 2 • It consisted by essentially of two
compartments C and D separated by a Sartorius cellulose acetate micro filtration membrane (cat No
11107, average pore size 0.2 )lm and thickness
1xl0-4m). An aqueous solution of lecithin cholesterol
mixture of fixed composition was added to compartment C of the transport cell along with the aqueo us
NOTES
solution of lecithin cholesterol mixture of the same
composition in compartment D. The pH of the solution was maintained at 7.26 in both compartments
using appropriate buffers. For maintaining pH a phosphate buffer was used. The concentration of lecithin
cholesterol mixtures used in experiment was 1.919 x
10-5 M with respect to lecithin and 1.175 x 10--{j M
with respect to cholesterol that liquid membrane generated by lecithin at the interface is completely saturated with cholesterol and control value was noted.
The concentration of bile salts was 0.272 gm in 100
ml of lecithin cholesterol mixture and the drug was
0.5% aqueous solution in lecithine cholesterol and
bile salt mixture. To obtain data on hydraulic permeability known pressure were applied on compartment
'C' and the consequent volume flux was measured by
noting the rate of advancement of liquid meniscus in
the capillary attached to compartment 'D'.
~he experiment was also performed by taking bile
salts and drugs in other compartment. All measurements were made at constant temperature using thermostat set at 37°± 1°C. The data obtained on hydraulic
permeability were calculated as :
JV
= r2 i/r2 t = (r2 )/rl/t
JV
=volume flux
R
= Radii of capillary L 2
r
= Radii of capillary L 1
I
=Distance
t
=Time
JV
= Lp X ~p
Where Lp =Hydraulic conductivity
~p = Pressure difference
The value of hydraulic permeability was expressed
in following unit;
w·s M 3s -I N·I
The statistical calculation was done by student's 't'
test.
The hydraulic permeability of membrane in the
presence of indigenous drugs was recorded. A significant decrease in the value of hydraulic permeability
was observed for all the indigenous drugs tested as
compared with controlled value (0.49 x 10- 8 M- 3 S- 1
W 1) (P = <0.02, P < 0.01). The maximum decrease in
the value of hydraulic permeability was recorded for
Tinospora cord~folia (Guduchi) as compared with
other drugs (P < 0.001) (Table).
Srivastava2 and others have already explained that
the liquid membrane, developed by lecithin and cholesterol reduces both fluxes, through the supporting
membrane, namely that of salt and water 7. 10 . In the
present experimental study it was observed that hy-
1309
Table !-Hydraulic permeability of membrane in presence of bile
salt and indigenous drugs
[Values are mean± SE]
Combinations
Lpx i0- 8 M 3
s - IN- 1
I.
Lecithin+ cholesterol (control)
0.49 ± 0.015
2.
Lecithin cholesterol + bile salt
(sodium deoxycholate)
Lecithin + cholesterol + bile salt+
daruharidra
Lecithin + cholesterol + bile salt + kutki
0.47 ± 0.011 a
Lecithin + cholesterol + bi le salt +
bhoomyamalaki
Lecithin+ cholesterol + bile salt+
kalmegha
Lecithin +cholesterol + bile salt+
guduchi
Lecithin +cholesterol+ bile salt+
all above mentioned Indigenous drugs
in combined form
0.29 ± 0.019b
Sl.
No.
3.
4.
5.
6.
7.
8.
0.45 ± o.o25•
0.30 ± 0.027 b
0.28 ± 0.020b
0.27 ± O.OII c
0.30 ± 0.020b
P values: a <0.02; b < 0.0 I; c <0.00 I.
draulic permeability of lipid membranes decreases in
the presence of bile salt (sodium deoxycholate) and a
further decrease was observed in the presence of bile
salt and Indigenous drugs. This mechanism of action
of drugs to decrease the hydraulic permeability in the
presence of bile salt can be described for hepatic cells
which bear inflammation and destruction in case of
hepatitis. Thus, the decrease in hydraulic permeability
in the presence of these drugs could prevent the inflammatory process and may cheque the complications caused by bilirubin as a result of destruction of
hepatic cells. It has been observed that altered permeability of membrane due to inflammation and infection affects the hydraulic permeability, which is further altered in the presence of bile salt and bile pigment11. It appears plausible that these indigenous
drugs have a membrane stabilizing property and may
prevent the toxic effect of bile salt by improving the
biliary flow amongst hepatic and biliary canaliculi.
On the basis of the present results, it can be concluded that the used indigenous drugs are having
hepato-protective as well as cell stabilizing property .
Although all the used indigenous drugs decreased the
hydraulic permeability significantly but the drug Tinospora cordifolia (Guduchi) showed the maximum
decrease in the hydraulic permeability as compared
with separate as well as combine form of all the drugs
used in this study.
1310
INDIAN 1 EXP BIOL, DECEMBER 2001
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