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Potential for Pakistani traditional medicinal plants to combat diabetes

Article in Journal of Traditional Chinese Medicine · August 2014

DOI: 10.1016/S0254-6272(15)30051-0 · Source: PubMed

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Online Submissions: http://www.journaltcm.com J Tradit Chin Med 2014 August 15; 34(4): 488-490
info@journaltcm.com ISSN 0255-2922
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EXPERIMENTAL STUDY
TOPIC

Potential for Pakistani traditional medicinal plants to combat diabe-

tes

Mashwani Zia-ur-rehman, Khan Mirajab, Ahmad Mushtaq

aa

Mashwani Zia-ur-rehman, Department of Botany, Pir Mehr CONCLUSION: These findings will be helpful for fur-
Ali Shah Arid Agriculture University, Rawalpindi 46300, Paki- ther research into the use of traditional herbal med-
stan; Department of Plant Sciences, Quaid-i-Azam Universi- icines with antiglycation properties in the treat-
ty, Islamabad 45320, Pakistan
ment of diabetes.
Khan Mirajab, Ahmad Mushtaq, Department of Plant Sci-
ences, Quaid-i-Azam University, Islamabad 45320, Pakistan
Correspondence to: Mashwani Zia-ur-rehman, Depart- © 2014 JTCM. All rights reserved.
ment of Botany, Pir Mehr Ali Shah Arid Agriculture Universi-
ty, Rawalpindi 46300, Pakistan; Department of Plant Scienc- Key words: Diabetes mellitus; Plants, medicinal;
es, Quaid-i-Azam University, Islamabad 45320, Pakistan. zia.
Medicine, traditional; Antiglycation
botany@gmail.com; mashwani@uaar.edu.pk
Telephone: +92-333-9022077
Accepted: December 3, 2013
INTRODUCTION
Diabetes mellitus is a serious metabolic disorder with
multiple complications. The role of free radicals in dia-
betes has been widely discussed,1 and the involvement
Abstract of free radicals in diabetes onset and complications has
OBJECTIVE: To investigate the potential of medici- been shown experimentally.2 Hyperglycemia is the dis-
nal plants used in traditional healing in Pakistan to tinguishing feature of diabetes, and persistent condi-
inhibit the glycation process. This may be useful in tions in a diabetic patient lead to the formation of oxi-
combatting diabetes mellitus and its complica- dative stress for multiple reasons, including the autoxi-
tions. dation of glucose. This autoxidation generates free radi-
cals, including hydrogen peroxide, which is highly reac-
METHODS: The in vitro antiglycation activity of 10 tive after interaction with biomolecules, and accelerates
medicinal plants was examined by testing the abili- the formation of advanced glycation end products (AG-
Es). These AGEs accumulate in tissues, and crosslink-
ty of the extracts to inhibit the methyl-glyoxal-me-
ing with other macromolecules results in abnormalities
diated development of fluorescence of bovine se-
in cellular and tissue function.3 At present, the treat-
rum albumin.
ment of diabetes involves various therapeutic agents in
addition to insulin. However, because of adverse ef-
RESULTS: Of the tested plants, Persicaria barbata, fects and variable efficacy of these agents, there is inter-
Geranium collinum and Berberis lycium showed est in alternative treatment options.4 Therefore, plants
significant inhibition of the formation of advanced are considered to be an unexplored and rich source of
glycation end products. The inhibitory capacity of potential new antidiabetic agents. However, there is a
these plants was 68.89% , 62.06% and 54.23% , re- lack of mechanism-based in vitro assays, and only a few
spectively, compared with the positive control (ru- plants have been subject to thorough investigation.5-7
tin; 86% ). All other plants inhibited AGE formation In recent years, there has been an increasing trend to
non-significantly. investigate traditional medicinal plants for their poten-

JTCM | www. journaltcm. com 488 August 15, 2014 | Volume 34 | Issue 4 |
 Zia-ur-rehman M et al. / Experimental Study

tial as antidiabetic agents. Various plants have been et al.18 All the assays were performed in 96-well microti-
studied to date include Costus pictus,8 Eulophiaochrea- ter plate. The reaction mixture was prepared and 60 µL
ta,9 Microcepha lalamelatta,10 Dendrobium aqueum,11 placed in each well. The reaction mixture comprised of
Salvia sahendica,12 Berginia,13 Enicostema axillare,14 three components: 20 µL of 50 mg/mL glucose and
Rhynchosia reniformis,15 and Aralia taibaiensis.16 The 14 mM magnesium oxide, 20 µL of 10 mg/mL BSA,
objective of the study was to screen plants with antigly- and 20 µL of the plant extract. Blanks comprised of
cation properties for the treatment of diabetes. 60 µL with 40 µL of sodium phosphate buffer and
20 µL of BSA, and the negative control contained
20 µL of 0.1 M sodium phosphate buffer, 20 µL of
MATERIALS AND METHODS BSA and 20 µL of 30 mM NaN3. The plate was incu-
bated for 9 days at 37℃. Following incubation, 60 µL
Plants collection of 100% TCA was added to each well, followed by cen-
The plant specimens were collected from Galliyat re- trifugation at 15 000 rpm at 4℃ for 4 min. Next, 5%
gion, Western Himalaya, Pakistan. Identification of TCA was used to wash the pellets, containing the AGEs
specimens was performed by Plant taxonomy working bound with BSA, after that it was dissolved in phosphate
group at Plant Taxonomy and Biosystematics Labora- buffer solution (60 µL). The supernatant, which includ-
tory, Department of Plant Sciences, Quaid-i-Azam ed extraneous substances, glucose and inhibitors, was dis-
University (QAU), Islamabad. The plants were Gerani- carded. AGE formation was visualized fluorescence in-
um collinum, Persicariabarbata, Impatiens edgeworthi, tensity excitation (370 nm) and emission (440 nm) by
Clematis grata, Geranium wallichianum, Berberis lyci- using spectrofluorometer. The percentage inhibition of
um, Artemisia vulgaris, Rubia cordifolia, Boerhavia AGEs was determined as follows:
procumbens, and Capsella bursa-pastoris. Percentage inhibition=1 − (absorbance extract/absorbance con-
trol)×100
Extraction The results were interpreted according to the percent
Fresh plant material of 100 g was collected for each inhibition of AGE's. Values more than 80% were con-
specimen and under the shade dried and then grind to sidered as very significant, between 50%-80% were sig-
obtain the powdered samples. This powder is then nificant, 30%-50% were less significant and below that
used for the extract preparations. Powder was added were non-significant.
with 95% pure methanol (Sigma Co., St. Louis, MO,
USA) at room temperature and placed for three days
and then whole the process was repeated. The extracts RESULTS
obtained were filtered using Whatman filter paper 1
The antiglycation capabilities of the medicinal plants
(Sigma Co., St. Louis, MO, USA) and then filtrates
were investigated for their potential to inhibit of the
were pooled together. The pooled filtrate was concen-
formation of AGEs.
trated with rotary evaporator (Laborota 4000, Heido-
Rutin, which was used as a reference drug, showed
lph, Germany) at 40℃ under lowered pressure. Final
86% inhibition. The degree of antiglycation activity
dried and concentrated extract was weighed. Which
varied considerably from plant-to-plant. Persicariabar-
were stored in refrigerator for further processing in as-
bata, Geranium collinum and Berberis lycium showed
say. Rutin (Flex Pharma, Lahore, PK) was used as a
significant inhibition of AGE formation. The inhibito-
positive control.
ry capacity for these plants was 68.89% , 62.06% and
54.23% , respectively. All other plants inhibited AGE
Materials
formation non-significantly. These included Impatiens
The materials used were as follows: Bovine serum albu-
edgeworthi (17.15%), Clematis grata (9.43%), Gerani-
min (BSA; Merck Co., Darmstadt, Germany), 96 well
um wallichianum (3.17% ), Artemisia vulgaris
microtiter plates (EMD Millipore Corporation, Billeri-
(13.9%), Rubiacordifolia (17.74%), Boerhaviaprocum-
ca, MA, USA), glucose anhydrous (Sigma Co., St. Lou-
bens (9.52%), and Capsella bursa-pastoris (5.87%).
is, MO, USA), magnesium oxide (Merck Co., Darm-
stadt, Germany), sodium phosphate buffer (Merck
Co., Darmstadt, Germany), NaN3 (Wako Co., Osaka,
Japan), trichloroacetic acid (TCA; Sigma Co., Germa-
DISCUSSION
ny), and a spectrofluorometer (Shimadzu Japan). Of all plants tested in our study, three showed signifi-
cant inhibition of AGE formation. These were Persicar-
Experimental procedure iabarbata, Geranium collinum and Berberislycium.
The in vitro antiglycation activity of the medicinal No previous reports were found on the antiglycation
plants was examined by testing the ability of the ex- activities of the plants used in our study in the litera-
tracts to inhibit the methyl-glyoxal-mediated develop- ture. While the methanolic extract of Microcephala la-
ment of the fluorescence of BSA.17 The assay was per- mellate showed antiglycation activity 62.84% 10 and
formed according to the modified method of Kazeem 50% of AGE's inhibition was shown by Salvia officina-

JTCM | www. journaltcm. com 489 August 15, 2014 | Volume 34 | Issue 4 |
 Zia-ur-rehman M et al. / Experimental Study

lis at 0.62 mg/mL concentration, Petroselinum crispum ylase inhibitory potential of Eulophiaochreata L. J Pharm
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