Effect of Majun Baladur on life span, climbing ability, oxidative stress and dopaminergic neurons in the transgenic Drosophila model of Parkinson's disease

Abstract

The effect of a poly herbal drug Majun Baladur (MB) was studied on the transgenic Drosophila melanogaster expressing human alpha synuclein in the neurons (PD flies). The equivalents of recommended dose for human were established for 20 g of fly food i.e. 0.0014, 0.0028, 0.0042 and 0.0056 g per 20 g of diet. The PD flies were allowed to feed on it for 24 days before performing the assays. The exposure to MB increased the life span and improves the activity of PD flies. The PD flies exposed to 0.0014, 0.0028, 0.042 and 0.0056 g of MB showed a dose dependent significant delay of 1.47, 1.88, 2.52 and 3.05 folds in the climbing ability compared to unexposed PD flies. A dose dependent significant decrease of 1.38, 1.45, 1.48 and 1.65 folds in TBARS; 1.08, 1.11, 1.17 and 1.20 folds in the GST activity; 1.20, 1.28, 1.39 and 1.52 folds in the PC content; 1.43, 1.53, 1.65 and 1.79 folds in the Caspase-9 activity; 1.21, 1.31, 1.53 and 1.64 folds in the activity of Caspase-3 and 1.24, 1.42, 1.50 and 1.79 folds in the activity of catalase; 1.50, 1.63, 1.88 and 2.06 folds in the activity of SOD in PD flies exposed to 0.0014, 0.0028, 0.042 and 0.0056 g of MB, respectively. A significant dose dependent increase of 1.20, 1.29, 1.33 and 1.44 folds in as NPSH content was observed in PD flies exposed to 0.0014, 0.0028, 0.042 and 0.0056 g of MB, respectively. The exposure to MB protects the loss of dopaminergic neurons as is evident by immunohistochemistry. It is concluded that MB is potent in reducing the PD symptoms being mimicked in the transgenic flies.

Keywords: Neuroscience.

Fig. 1

Climbing ability in Parkinson's disease (PD) flies and control for a period of 24 days. The values are mean of five assays.

Fig. 2

Effect of Majun Baladur (MB) on the climbing ability activity of flies. [MB1 = 0.0014 g; MB2 = 0.0028 g; MB3 = 0.0042 g; MB4 = 0.0056 g; PD = PD flies; Dopamine = 10⁻³ M; N = 50]. The flies were allowed to feed on the diet supplemented with Majun Baladur for 24 days and then assayed for climbing ability activity. [The doses were established per 20g of diet; ^asignificant difference with respect to control, p < 0.05; ^bsignificant difference with respect to PD flies p < 0.05].

Fig. 3

(a) and (b) shows the average activity pattern and chi-square periodogram, respectively for control flies (N = 20).

Fig. 4

(a) and (b) shows the average activity pattern and chi-square periodogram, respectively for PD flies (N = 20).

Fig. 5

(a) and (b) shows the average activity pattern and chi-square periodogram, respectively for PD flies exposed to polyherbal drug Baladur (0.0014 g). [The doses were established per 20 g of diet].

Fig. 6

(a) and (b) shows the average activity pattern and chi-square periodogram, respectively for PD flies exposed to polyherbal drug Baladur (0.0028 g). [The doses were established per 20 g of diet].

Fig. 7

(a) and (b) shows the average activity pattern and chi-square periodogram, respectively for PD flies exposed to polyherbal drug Baladur (0.0042 g). [The doses were established per 20 g of diet].

Fig. 8

(a) and (b) shows the average activity pattern and chi-square periodogram, respectively for PD flies exposed to polyherbal drug Baladur (0.0056 g). [The doses were established per 20 g of diet].

Fig. 9

(a) and (b) shows the average activity pattern and chi-square periodogram, respectively for PD flies exposed to dopamine (10⁻³ M).

Fig. 10

(a) and (b) shows the average activity pattern and chi-square periodogram, respectively for control flies exposed to polyherbal drug Baladur (0.0014 g).[The doses were established per 20 g of diet].

Fig. 11

(a) and (b) shows the average activity pattern and chi-square periodogram, respectively for control flies exposed to polyherbal drug Baladur (0.0028 g).[The doses were established per 20 g of diet].

Fig. 12

(a) and (b) shows the average activity pattern and chi-square periodogram, respectively for control flies exposed to polyherbal drug Baladur (0.0042 g).[The doses were established per 20 g of diet].

Fig. 13

(a) and (b) shows the average activity pattern and chi-square periodogram, respectively for control flies exposed to polyherbal drug Baladur (0.0056 g).[The doses were established per 20 g of diet].

Fig. 14

Effect of Majun Baladur (MB) on the survival rate. [MB1 = 0.0014 g; MB2 = 0.0028 g; B3 = 0.0042 g; B4 = 0.0056 g; PD = PD flies; Dopamine = 10⁻³ M; N = 50]. The values are the mean of 5 assays.

Fig. 15

Effect of Majun Baladur (MB) on the thiobarbituric acid reactive species (TBARS) in the brains of flies. [MB1 = 0.0014 g; MB2 = 0.0028 g; MB3 = 0.0042 g; MB4 = 0.0056 g; PD = PD flies; Dopamine = 10⁻³ M; N = 50]. The flies were allowed to feed on the diet supplemented with Majun Baladur for 24 days and then assayed for lipid peroxidation. [The doses were established per 20 g of diet; ^asignificant difference with respect to control, p < 0.05; ^bsignificant difference with respect to PD flies p < 0.05].

Fig. 16

Effect of Majun Baladur (MB) on the glutathione-S-transferase (GST) activity in the brains of flies. [MB1 = 0.0014 g; MB2 = 0.0028 g; MB3 = 0.0042 g; MB4 = 0.0056 g; PD = PD flies; Dopamine = 10⁻³ M; N = 50]. The flies were allowed to feed on the diet supplemented with Majun Baladur for 24 days and then assayed for GST activity. [The doses were established per 20 g of diet; ^asignificant difference with respect to control, p < 0.05; ^bsignificant difference with respect to PD flies p < 0.05].

Fig. 17

Effect of Majun Baladur (MB) on the Non-protein thiol (NPSH) content in the brains of flies. [MB1 = 0.0014 g; MB2 = 0.0028 g; MB3 = 0.0042 g; MB4 = 0.0056 g; PD = PD flies; Dopamine = 10⁻³ M; N = 50]. The flies were allowed to feed on the diet supplemented with Majun Baladur for 24 days and then assayed for NPSH content. [The doses were established per 20 g of diet; ^asignificant difference with respect to control, p < 0.05; ^bsignificant difference with respect to PD flies p < 0.05].

Fig. 18

Effect of Majun Baladur (MB) on the protein carbonyl content in the brains of flies. [MB1 = 0.0014 g; MB2 = 0.0028 g; MB3 = 0.0042 g; MB4 = 0.0056 g; PD = PD flies; Dopamine = 10⁻³ M; N = 50]. The flies were allowed to feed on the diet supplemented with Majun Baladur for 24 days and then assayed for protein carbonyl content. [The doses were established per 20 g of diet; ^asignificant difference with respect to control, p < 0.05; ^bsignificant difference with respect to PD flies p < 0.05].

Fig. 19

Effect of Majun Baladur (MB) on the Caspase-9 activity in the brains of flies. [MB1 = 0.0014 g; MB2 = 0.0028 g; MB3 = 0.0042 g; MB4 = 0.0056 g; PD = PD flies; Dopamine = 10⁻³ M; N = 50]. The flies were allowed to feed on the diet supplemented with Majun Baladur for 24 days and then assayed for Caspase-9. [The doses were established per 20 g of diet; ^asignificant difference with respect to control, p < 0.05; ^bsignificant difference with respect to PD flies p < 0.05].

Fig. 20

Effect of Majun Baladur (MB) on the Caspase-3 activity in the brains of flies. [MB1 = 0.0014 g; MB2 = 0.0028 g; MB3 = 0.0042 g; MB4 = 0.0056 g; PD = PD flies; Dopamine = 10⁻³ M; N = 50]. The flies were allowed to feed on the diet supplemented with Majun Baladur for 24 days and then assayed for Caspase-3. [The doses were established per 20 g of diet; ^asignificant difference with respect to control, p < 0.05; ^bsignificant difference with respect to PD flies p < 0.05].

Fig. 21

Effect of Majun Baladur (MB) on the catalase in the brains of flies. [MB1 = 0.0014 g; MB2 = 0.0028 g; MB3 = 0.0042 g; MB4 = 0.0056 g; PD = PD flies; Dopamine = 10⁻³ M; N = 50]. The flies were allowed to feed on the diet supplemented with Majun Baladur for 24 days and then assayed for catalase. [The doses were established per 20 g of diet; ^asignificant difference with respect to control, p < 0.05; ^bsignificant difference with respect to PD flies p < 0.05].

Fig. 22

Effect of Majun Baladur (MB) on the Superoxide dismutase (SOD) content in the brains of flies. [MB1 = 0.0014 g; MB2 = 0.0028 g; MB3 = 0.0042 g; MB4 = 0.0056 g; PD = PD flies; Dopamine = 10⁻³ M; N = 50]. The flies were allowed to feed on the diet supplemented with Majun Baladur for 24 days and then assayed for Superoxide dismutase (SOD). [The doses were established per 20 g of diet; ^asignificant difference with respect to control, p < 0.05; ^bsignificant difference with respect to PD flies p < 0.05].

Fig. 23

Tyrosine hydroxylase (TH) immunostaining performed on the brain sections of flies after 24 days of the exposure; a-Control, b-PD fly, c-PD fly exposed to MB1 = 0.0014 g, d-PD fly exposed to MB2 = 0.0028 g, e-PD fly exposed to MB3 = 0.0042 g, f-PD fly exposed to MB4 = 0.0056 g, g-Dopaminergic neurons were quantified in terms of TH-positive cells from the total area of the brain by using Image J software (N = 5). [MB1 = 0.0014 g; MB2 = 0.0028 g; MB3 = 0.0042 g; MB4 = 0.0056 g; PD = PD flies.