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Indian Journal of Experimental Biology

Vol. 40, March 2002, pp. 273-276

Amelioration of experimental diabetic neuropathy and gastropathy in rats

following oral administration of plant (Eugenia jambolana, Mucurna pruriens
and Tinospora cordifolia) extracts
J K Grover*, S S Rathi & V Vats
De partments of Pharmacology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India
Received 11 Jun e 2001; revised 19 December 2001

Extract of M. charanlia (200 mg/kg), £. jambolana (200 mg/kg), M. pruriens (200mg/kg) and T. cordi/alia (400 mg/kg)
was administered for 50 days in STZ induced diabetic mice, the plasma glucose concentration was reduced by 24.4, 20.84,
7.45 and 9.07 % respectively. Tail nick latency (TFL) and gastric transit percentage were significantly higher in diabetic
controls versus normal controls. M. charanlia and £. jambo/ana modifi ed it favorably while M. pruriens and T. co rdi/alia
did not exert any favorable change.

Diabetic neuropathy is the complication of diabetes Anti-hyperglycemic activity of M. charantia 8, E.

mellitus and occurs in almost 50% of diabetic jambolana 9 , M. pruriens lo and T. cordifolia" has been
patients I . Patients with autonomic neuropathy are reported earlier but none of these studies have
characterized by maladaptation to everyday stress revealed the effect of plant extracts on development
situations like exercise and hypoglycemia which of diabetic complications such as neuropathy and
gastropat hy. In our prevIOus stu d·les 1213
. , M . charantta
predispose them to a higher risk of death 2.
Gastrointestinal symptoms are also responsible for and E. jambolana have shown higher anti-
substantial morbidit/ A and approximately 30 to 50 % hyperglycemic effect as compared to T. cordifolia and
M. pruriens in mild (plasma sugar> 180 mg/dl ,
of patients with long-standing diabetes mellitus have
duration 21 days), moderate (plasma sugar> 280
delayed gastric emptying time and up to 60% have
mg/dl, duration 120 days) and severe (plasma
severe intestinal symptoms like nausea, vomiting,
sugar> 400 mg/dl, duration 60 days) diabetic rats. In
diarrhoea and constipation 3 . Presently, other than addition, M. charantia and E. jambolana also partially
symptomatic control no effecti ve treatment is restored altered hepatic and skeletal muscle glycogen
available though metabolic control has been shown to content and hepatic glucokinase, hexokinase, glucose-
strongly influence the development of diabetic 6-phosphate and phosphofructokinase levels.
neuropathy and gastroparesis4 . In the recent years, Moreo~er, E. jambolana and M. charantia also
popularity of complementary medicine has increased attenuated certain parameters in STZ induced diabetic
as shown by surveys conducted in Australia and US, nephropathyl 4 and dietary fructose 'induced insulin
which indicate that almost 48.5 and 34 % respondents resistance in normal rats IS .
had used at least one form of unconventional therapy
including herbal medicine s.6 . Dietary measures and Materials and Methods
traditional plant therapies prescribed by Ayurveda and Extract of plant parts-Fresh green fruit of
other indigenous systems of medicine have been used Momordica charantia and dried kernels of ripen
commonly in India7 . In the present study, 4 plants Eugenia jambolana were collected locally . Cuticle of
namely Momordica charantia ; Eugenia jambolana ; M. charantia fruit was peeled off and macerated in an
Mucuna pruriens; and Tinospora cordifolia with electric mixer (Electro Com, New Delhi), the kernels
previously confirmed anti-hyperglycemic activity of E. jambolana were grounded in an electric grinder.
were selected for the present study. The macerated pulp of M. charantia and the E.
jambolana powder were then soaked separately in
*Correspondent author- Fax: 91-11-6862663; equal amount of water and stirred intermittently and
Email: jkgrover @hotmail.com then left overnight. This pulp was then filtered

through a coarse sieve and the filtrate was dried at PO every day till the end of experiment (up to day
reduced temperature. Respective yield of M. 50). Animals described as fasting had been deprived
charantia and E. jambolana was 80.4 gmlkg of fruit of food for at least 16 hr but given water ad libitum.
and 73 gmlkg of kernel. To increase the shelf life and Plasma glucose-Glucose levels were estimated
uniformity, the extracts were completely lyophilized by commercially available glucose kits based on
by continuous freeze drying operation of 54 hr (Christ glucose oxidase method 16 on every 10 day for 50 days
freeze dryer, alpha 1-4, Germany), yielding 64 and 51 (Autopak®, Bayer Diagnostics, Baroda).
g/ lOOg of M. charantia and E. jambolana extract Tail flick latency (TFL)-On day 50 of the
respectively. experiment, TFL was studied as a parameter to assess
Alcoholic and aqueous ex tracts of T. cordiJolia and diabetic neuropathy on electrical analgesiometer
M. pruriens were received as gift from Brawn (Techno, India), before and after morphine injection
Pharmaceuticals LTD, Faridabad (India). All the (5mg/kg; sc) at 30 and 60 mi n intervals l7 . Maximum
extracts were suspended in 1% carboxy latency period for the mice to flick its tail was fixed at
methylcellulose (Central Drug House, New Delhi) 15 sec lest the mouse burnt its tail. Index of analgesia
and given orally. (%) in mice was calculated by the following formula:
Sample collection-Blood was collected retro- Index of analgesia (%) = Test Latency x 100! Basal
orbitally from the inner canthus of the eye under light Latency
ether anesthesia using capillary tubes (Micro Rate of transit of charcoal in CIT-It was
Hematocrit Capillaries, Mucaps). Blood was collected determined on day 50 of the experiment by forced
in fresh vials containing sodium fluoride and sodium feeding of charcoal (12.5 %) and gum acacia (12.5
oxalate as anti-coagulantlanti-glycolytic agents and %) 18. Ten min after feeding charcoal, animals were
plas ma was separated in a T8 electric centrifuger sacrificed and intestine was removed. Total length of
(Remi Udyog, New Delhi) at 2000 rpm for 2 min. intestine and distance travelled by charcoal in the
Preparation of diabetic animals-Male albino intestine were measured and GTT was calculated as
mice (20-35g) were fasted for overnight and given a distance travelled by charcoal in the intestine divided
bolus injection of STZ (150 mg/kg ; ip) dissolved in by total length of intestine.
citrate buffer (3mM ; pH 4.5). After 10 days, serum Statistical analysis-The results were analyzed for
glucose was estimated using commercially available statistical significance by Student's t test using
Autopak® kit, Bayer Diagnostics, Baroda. Mice computerised software, GraphPAD, InStat, 1990,
exhibiting plasma glucose levels > 300 mg/dl were Version 1.14, INSERM 920666S , India.
used for further study.
Experimental design-Albino mice (30-50g) of Results and Discussion
both sexes (8-10 weeks old) were obtained from the Aim of the present work was to study the effect of
Institute and maintained on standard chow diet during 50 days of persistent hyperglycemia on TFL
the experiment. The animals were randomized in the (neuropathy) and rate of transit of charcoal in GIT
following groups with a acclimatization period of 2-3 (gastropathy) and whether 4 plant extracts (M. charantia ;
days before initiation of experiment. Mice were E. jambolana; M. pruriens and T. cordiJolia ) modify
divided into 6 groups having 8 mice in each group. In the outcome favorably. In addition, modification of
group I, mice received saline daily and served as analgesic effect of morphine and glucose levels were
normal control. In group II to VI, mice received a also assessed in the same. Effect of administration of
single intra peritoneal injection of 150 mg/kg STZ. In plant extracts of M. charantia, E. jambolana, M.
group II, mice received saline daily and served as a pruriens and T. cordiJolia for 50 days in STZ diabetic
diabetic control. Group III received 200 mg of mice is given in Table 1 . Total reduction of blood
lyophilized powder of M. charantia, group IV glucose was of 24.4, 20.84, 9.07 and 7.45 %
received 200 mg of lyophilized powder of E. respectively.
jambolana, and group V received 400 mg of aqueous Effect of administration of plant extracts on rate of
extract of T. cordiJolia and group VI received transit of charcoal in GIT in STZ diabetic mice has
200 mg/day of alcoholic extract of M. pruriens. The been shown in Table 2. There was a significant
doses of the plant extracts were decided on the basis difference in mean rate of transit in diabetic control vs
of a previous stud/ 2,13. All the extracts were given normal control (83.00±6.38 vs 99.16± 1.32%

Table 1 - Effec t of plant extracts on g lucose (mg/%) le ve ls in streptozotoc in (l50/mg/kg) diabetic mi ce

[Values are mean±SD for 8 an imals in eac h group]
Group oday Day 10 Day 20 Day 30 Day 40 Day 50 Day 60

NC 96.3 ±3.9 94.98 ±4.67 95.5 ±4.85 95 .5±4.85 100.0 1 ±6.28 94.26± 4.38 92 .03 ±4.32
DC 90.3 ± 2. 1 437.05 ± 12.8** 44 1.1 ± 19.9** 471.3± 19.3** 470.5 ± 12.3** 450.5 ± 10.03** 439.5 ± 14.2**
MC 98.61 ±5.33 450.8 1 ± 13 .39 449.33 ± 22.33 418±22.08 398. 16± 12.44** 366± 14.61** 339.66± 14.22***
EJ 95.9±3.4 439.8±7.9 456.5 ±22 .9 466.6± 15.4 430.6± 18.8* 399.3 ± 29.4** 361.8 ± 3 1.1 ***
MP 95.21 ±2.97 437.61 ±9.8 1 464.66 ± 15.57 472±12.5 473 .83± 20.9 447±1 8.4 430± 14.97**
TC 99 .06±2.6 454.7 ± 11 .3 462.8± 14.4 482 .3± 8.5 462 .8 ±6.8 455.8± 12.9 420.8 ± 13.3*

., Diabetic control was co mpared with the normal, and experime ntal gro ups were compared with their own corresponding va lues on day 10.
NC- Non-di abeti c con trol; DC-diabetic control; MC-M. charantia; EJ -E. jambolana; MP-M . pruriens; TC - T. cordifolia.
Significant at *P<O .O I, **<0.001, ***<0.0001

Table 2-Effec t of STZ and plant ex tracts on gastrointest inal Tab le 3 - Effec ts of pl ant ex trac ts on TFL in mice (in sec)
transit time in mice [Val ues are mean ± SD for 8 animals in each group]
[V alues are mea n ± SD fo r 8 an imals in each group]
Groups o min 30 min 60 min
Groups Rate of transit of charcoal
in GIT (%) Control 6. 11 ± 1.09 9.01 ± 1.8 12.05 ± 0.7
Di abetic contro l 10.6 1 ± 0.9* 11.8 ± 1.6 12.7 ± 1.4
Normal 99. 16±1.32
M. charantia 7.95 ± 0.9 10.52 ± 1.2 13.4 1 ± 1.5
Di abetic Control 83.00±6.38**
£. jambolana 8.5 ± 1.3 9.85 ± 1.7 12.55 ± 1.3
M. charantia 90.28±5.4 1*
M. pruriens 8.92 ± 0.5 9.71 ± 1.2 12.0 ± 0.9
£. jambolana 90.20±5.20*
T. cordifolia 8.5 ± 1.3 11.55 ± 1.7 13.56 ± 1.3
M. pruriens 80.60±5.24
T. cordifolia 83.60±5.20 Di abetic control was compared with normal and ex perimental
groups were co mpared with di abetic control
Diabetic cont ro l was compared with the normal, and experimental Si gnificant at * P < 0.05
groups were compared with diabetic cont ro l
Significant at * P < 0.05 ; and **< 0.005 Thus, it is likely that M. charantia and E. jambolana
normalised rate of tran sit of charcoal in GIT through
respectively) . Th is finding is consistent with the
an unknown independent mech anism other than its
previous finding 19 that gastric emptying rate is faster
anti-hyperglycemic effect.
in hypoglycemic state and slower in case of
hyperglycemic state. Hyperglycemia 20 has been Effect of administration of plant extracts on TFL of
implicated as the main factor respon si ble for delay in STZ di abetic mice is shown in Table 3 and index of
gastric emptying though direct effects of metaboli c analgesia has been shown in Table 4 . TFL was
derangement via an altered autonomic function (i .e. significantly hi gher in diabetic mice (l 0.61 ± 2.98) as
neuropath y) cannot be ruled oue. M. charantia and E: compared to normal control (6.11 ± 1.09) befo re
jambolana caused a significant increase in gastric administration of morphine. These results are in
transit percentage with comparison to di abetic agreement with the previous finding s2 1-23 . However,
controls (90.28±5.41 and 90.20±5.20 versus decrease in hot plate latenc/ 4 as well as no change
83.00±6.38 respectively) . M. pruriens and T. has also been described earlier25 .26 . In the present
cordifolia had no such effect. Although M. charantia study, TFL in treated groups before the administration
and E. j ambolana caused a significant decrease in of morphine was insignificantly lesser than di abetic
serum sugar levels (24 and 21 % reduction controls. Daily admini stration of M. charantia and E.
respectively), an euglycemic state was not achieved. jambolana insignificantly prevented the rise in basal

Table 4- Effects of plant extracts on index of analgesia in mice II Wadood N, Wadood A & Shah S A, Effect of Tinospora
(i n % ) co rdifolia on blood glucose and total lipid levels of normal
and alloxan diabetic rabbits, Plal1la Med 58 (199 1) 131 .
[Values are mean ± SD for 8 animals in each gro up] 12 Grover J K, Vats V & Rathi S S, Anti-h ype rglyce mic cffects
Group 30 min 60 min of Eugenia jambolana and Tinospora cordifolia in
ex perimental diabetes and their effects on key metabolic
Control 147 .6 197.2 1 enzy mes involved in carbohydrate metabolism, J
Diabetic control 111.2 1 11 9.69 Ethnopharmaco/, 73 (2000) 461.
M. charantia 132.32 168.67 13 Rathi S S, Grover J K & Vats V, Anti-hyperglyce mic effects
E. jambo/ana 115 .88 147 .64 of Momordica charantia and Mu cul/a pru riens in
M. pruriens 108.85 134.52 ex perimental diabetes and their effect on key metabol ic
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Res, 200 I (in press).
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