ANTIANXIETY PROPERTY OF BASELLA ALBA LINN LEAVES EXTRACT IN EXPERIMENTAL ANIMALS - RESULTS

 

5. RESULTS

5.1. Successive Solvent Extraction:

The Basella alba leavespowder was subjected to batch extraction in Soxhlet apparatus. The percentage yield, colour, consistency and solubility in water were noted in table 1.

Table No. 1: The percentage yield, colour, consistency and solubility in water of Basella alba leaves extracts.

Plant part used	Extracts	% Yield	Colour	Consistency	Solubility in water
Basella alba leaves	Petroleum ether	5.45	Yellow	Sticky	Insoluble
	Chloroform	5.22	Dark Green	Semisolid	Slightly Soluble
	Ethanol	18.44	Dark brown	Semisolid	Freely soluble
	Aqueous	24.33	Dark brown	Semisolid	Freely soluble

 

5.2 Preliminary Phytochemical testing of Basella alba leaves extracts:

The investigation of the preliminary phytochemical qualitative examination of various extracts of Basella alba leavesshows the presence of different constituents in table. 2

 

 

Table No. 2: Preliminary Phytochemical testing of Basella alba leaves extracts:

Phytochemical constituents	Pet ether	Chloroform	Ethanol	Aqueous
Carbohydrates	+	++	+++	++
Proteins	-	-	-	-
Amino acid	-	-	-	-
Glycosides	+	+	+	++
Saponins	-	+	+	++
Flavonoids	-	-	+++	++
Alkaloids	-	+	+++	++
Tannins	-	-	++	+++
Steroids	+	+	-	-
Vitamins C&D	-	+	++	++

- Absent                      ++ more clarity

+ indicates                   +++ better response

 

5.3 Acute toxicity study:

Acute toxicity studies for aqueous and ethanolic extracts of Basella alba leaves were conducted as per OECD guidelines 425 using albino swiss mice. Each animal was administered aqueous and ethanolic extracts by oral route. The animals were observed for any changes continuously for the first 2 h and up to 24 h for mortality, there were no mortality and noticeable behavioural changes in all the groups tested. The extracts were found to be safe up to 2000 mg/kg body weight.

 

 

5.4 Elevated plus maze test:

5.4.1 Effects of aqueous extracts of Basella alba leaves on EPM:

The behavioural effects of diazepam and AEBAL at 400 & 800 mg/kg doses on the behaviour of mice in the elevated plus maze test were summarized in table 3 and figure 1 to 4.

Diazepam has increased the percentage of time spent in open arms significantly (p < 0.001, Figure 1) and of arm entries in open arms significantly (p < 0.01, Figure 3), whereas the percentage of time spent in closed arms has decreased significantly (p < 0.001, Figure 2) and of entries in closed arms has decreased significantly (p < 0.01, Figure 4) as compare to control group.

It was seen that the AE 400 mg/kg has no significant result but AE 800 mg/kg has increased percentage of time spent and of entries in open arm significantly (p < 0.05, Figure 1 and 3) whereas the percentage time spent in closed arm has decreased significantly (p < 0.05, Figure 2) but the percentage of entries in to closed arm has no significantly decreased as compare to control. The studies with that of AE 800 mg/kg shows significant results as compare to control group. AE 800 mg/kg has significantly increased the percentage time spent and arms entries in open arms (p < 0.01, Figure 1 and 3) whereas in closed arm it has decreased (p < 0.01, Figure 2 and 4) as compare to control group.

 

5.4.2 Effects of ethanolic extracts of Basella alba leaves on EPM:

The effects of EEBAL at EE 400&800 mg/kg and diazepam on the behaviour of mice that were summarized in table  and figures..

Diazepam has increased the percentage of time spent and of arm entries in open arms is significant (p < 0.001, Figure 1 and 3), whereas in closed arm it has decreased significantly (p < 0.01, Figure 3 and 4) as compare to control group.

The EE 400 mg/kg has shown insignificant in percentage of time spent and arm entry in open and closed. The EE 800 mg/kg has shown increase in percentage of time spent and of arm entries in open arms significantly (p < 0.05, Figure 1 and 3) and no significant decreased in time spent in closed arms (Figure 4) whereas significantly decreased in percentage of arm entries in the closed arms (p < 0.05  Figure 4) as compare to control group.

           The EE 800 mg/kg has increased the percentage of time spent and of arm entries in open arms was significantly (p < 0.01, Figure 1 and 3), whereas the both percentage of spent time and arm entries in closed arms were decreased significantly (p < 0.05, Figure 2 and 4).  

 

Treatment	% open arm time (s)	% closed arm time (s)	% open arm entries	% closed arm entries
Vehicle	12.17±1.721	77.67±3.252	23..00±1.265	73.67±3.921
Diazepam 2mg/kg	70.677±3.373***	23.17±1.628***	63.33±2.418***	28.83±1.108***
AE 400 mg/kg	23.50±1.628*	64.67±1.1.358 *	30.00±2.418	53.33±2.076*
AE 800 mg/kg	34.50±1.544**	45.50±4.380**	43.00±3.406*	39.67±1.382**
EE 400 mg/kg	33.66±2.124**	47.33±4.470**	49.67±0.881**	40.00±4.187***
EE 800 mg/kg	46.17±2.227***	33.67±2.565***	59.00±1.065***	32.33±2.459***

    Table No.3: Effects of Extracts of Basella alba leaves from elevated plus maze test in mice.

 

     All values are mean ± SEM, (n =6), one way ANOVA, followed by Dunnet’s test.

     * p < 0.05, ** p < 0.01, *** p < 0.001, when compared to vehicle treated group.

Figure 1 - Percentage of open arm time in 5-min EPM.

Figure 2- Percentage of closed arm time in 5-min EPM.

Figure 3- Percentage of open arm entry in 5-min EPM.

Figure 4- Percentage of closed arm entry in 5-min EPM.

 

5.5    Hole and board test:

5.5.1 Effects of aqueous extract of Basella alba leaves on hole and board test:

The effects of AE 400 & 800 mg/kg and diazepam were summarized in table 5 and figure 5 to 7.

            Diazepam revealed that, significantly decrease head dip latency (p < 0.001, Figure 9) and locomotion (p < 0.001, Figure 11) and increase head dip count (p < 0.001, Figure 10).

Further analysis shown that AE 400 mg/kg was insignificant in above paradigms, whereas AE 800 mg/kg shown decreased head dip latency significantly (p < 0.01, Figure 5) and increase head dip count (p < 0.01, Figure 6). The locomotion showed insignificant result as compare to control group.

The AE 800 mg/kg has decreased head dip latency significantly (p < 0.001, Figure 5) and increase head dip count (p < 0.001, Figure 10) whereas the locomotion shown decreased significantly (p < 0.01, Figure 7) as compare to control group.

 

5.5.2 Effects of ethanolic extract of Basella alba leaveson Hole and board test:

The effects of EE 400 & 800 mg/kg and diazepam were summarized in table 6S and figure 5 to 7. Diazepam exhibit that, significantly decreased head dip latency (p < 0.01, Figure 12) and increased head dip count (p < 0.001, Figure 6) whereas the locomotion decreased significant (p < 0.01, Figure 7).

Further analysis shown that EE 400 mg/kg has significant in above paradigms whereas EE 800 mg/kg shown decreased head dip latency significantly (p < 0.05, Figure 12), increased head dip count (p < 0.05, Figure 13) and locomotion decreased significantly (p < 0.05, Figure 14) as compare to control group.

The EE 400 mg/kg has shown decreased head dip latency significantly (p < 0.01, Figure 12), increased head dip count (p < 0.001, Figure 13) and locomotion decreased significantly (p < 0.01, Figure 7) as compare to control group.

 Table No. 4: Effects of Extract of Basella alba leaves from hole and board test in rats.

Treatment	Head Dip latency(s)	Head dip count	Locomotion(s)
Vehicle	68.17±2.762	20.17±1.515***	113.00±3.00
Diazepam 2mg/kg	12.00±1.483***	23.67±2.017	53.33±2.404***
AE 400 mg/kg	59.67±1.606*	23.67±1.430**	98.33±3.180*
AE 800 mg/kg	49.00±1.592**	33.17±1.662**	80.33±1.453**
EE 400 mg/kg	42.67±1.282**	40.00±1.291**	87.67±1.498**
EE 800 mg/kg	21.83±1.046***	51.17±1.222***	59.00±1.983***

 

                     All Values are mean ± SEM, (n =6), one way ANOVA, followed by Dunnet’s test.        

* p < 0.05, ** p < 0.01, *** p < 0.001 when compared to vehicle treated group.     

5.6 The Light and Dark transition test:

5.6.1 Effects of aqueous extract of Basella alba leaves on Light and Dark test:

Result of the light and dark transition test is depicted in table 5 and figure 8 to 10. Diazepam treatment group shown latency to enter in dark compartment significantly increased (p < 0.001, Figure 15) and  significantly increased time in light area (p < 0.01, Figure 16), and number of tunnel crossing was decreased significantly (p < 0.01, Figure 17).

The AE 400 mg/kg has not significant in above paradigms as compare to control group, whereas AE 800 mg/kg shown the latency to enter in dark compartment increased significantly  (p < 0.05, Figure 8) and  significantly increased time in light area (p < 0.05, Figure 16), and number of tunnel crossing was decreased significantly (p < 0.05, Figure 17).

The AE 800 mg/kg has shown increased latency to enter in dark significantly (p < 0.01, Figure 15) and  significantly increased time in light area (p < 0.01, Figure 16), and number of tunnel crossing was decreased significantly (p < 0.01, Figure 17).

 

5.6.2 Effects of Ethanolic extract of Basella alba leaves on light and dark transition test:

Result obtained from the light and dark transition test shown in table 5 and figure 8 to 10. Diazepam treatment group shown significantly increase latency to enter in dark compartment (p < 0.001, Figure 8) and time in light area (p < 0.01, Figure 19), whereas the number of tunnel crossing was decrease significantly (p < 0.001, Figure 20).

           Further analysis showed that EE 400 mg/kg has not significant in above paradigms. The EE 800 mg/kg was shown significantly increase the latency to enter in the dark chamber (p < 0.05, Figure 8), the time in light area (p < 0.05, Figure 9),whereas the number of tunnel crossing was decrease significantly (p < 0.05, Figure 10) as compare to control group.

The EE 400 mg/kg has shown significantly increase the latency to enter in the dark chamber (p < 0.01, Figure 8), the time in light area (p < 0.01, Figure 9),whereas the number of tunnel crossing was decrease significantly (p < 0.01, Figure 10) as compare to control group.

      Table No. 5: Effects of Extract of Basella alba leaves from light and dark transition test in mice.

Treatment	Latency to enter in dark area	Time spent in light area	Tunnel crossing
Vehicle	22.33±1.585	84.50±2.172	36.83±2.257
Diazepam 2mg/kg	53.17±3.609***	186.5±3.117***	16.00±1.592***
AE 400 mg/kg	27.17±2.072	102.7±3.879**	27.67±1.892**
AE 800 mg/kg	36.83±1.579**	143.00±3.276***	22.67±1.229**
EE 400 mg/kg	34.00±1.633**	119.5±2.884**	20.50±1.455**
EE 800 mg/kg	48.00±1.065***	175.5±3.713***	18.00±1.461***

 

        All values are mean ± SEM, (n =6), one way ANOVA, followed by Dunnet’s test.

        * p < 0.05, ** p < 0.01, *** p < 0.001 when compared to vehicle treated group.

5.7. Open field test:

5.7.1 Effects of aqueous extract of Basella alba leaves on OF test:

The effects of AEBAL doses as AE 400 & 800 mg/kg and diazepam on Open field test are shown in table 96 and figure 21 and 22. Diazepam has increase in locomotion (sec) significantly (p < 0.001, Figure 21) and decrease Rearing and Immobility (p < 0.01, Figure 22) as compare to control group..

The AE 400 mg/kg has not shown significant result  as compare to control group. The AE 800 mg/kg has shown increased locomotion significantly (p < 0.01, Figure 21). The AE 800 mg/kg has shown decreased Rearing and Immobility significantly (p < 0.001, Figure 21).

 

5.7.2 Effects of ethanolic extract of Basella alba leaveson O.F. test:

The effects of EEBAL at EE 400 & 800 mg/kg and diazepam on Open field  test were shown in table 6. Diazepam has increase in social interaction time (sec) significantly (p < 0.001, Figure 23) and decrease locomotor activity significantly            (p < 0.01, Figure 24).

       Table No. 6: Effects of Extract of Basella alba leaves from Open field test in rats.

Treatment	Total locomotion	Rearing	Immobility
Vehicle	105.2±2.182	33.00±2.206	25.50±1.432
Diazepam 2mg/kg	206.5±1.276***	9.167±1.276***	9.500±1.335***
AE 400 mg/kg	122.2±1.493*	26.67±1.726*	17.67±1.820**
AE 800 mg/kg	143.3±2.231**	21.83±1.558**	15.67±1.498***
EE 400 mg/kg	133.5±2.617*	25.17±1.167**	18.83±1.424*
EE 800 mg/kg	186.3±2.951***	12.50±1.544***	11.00±1.317***

 

         All values are mean ± SEM, (n = 3), one way ANOVA, followed by Dunnet’s test.

         * p < 0.05, ** p < 0.01, *** p < 0.001 when compared to vehicle treated group.