Studies on the Appetizing Effect of Apazin^®

 

Abstract:

 

  5g, 2.5g and 1.25g/kg (weight) of Apazin^® , equivalent to 38.46g, 19.23g and 9.62g respectively of crude drugs, which was about 16.44, 8.22, and 4.11 times the clinical dosage for adults, were given (ig) to mice for 12-14 days. The results show that Apazin^® compound has increased the food consumption of mice, enhanced the moving distance of activated char powder and the rate of the moving distance, and promoted the phenol red excretion of gastrointestinal tract. Compared with the control group, there was a significant difference. It has been demonstrated that the extract can affect the appetite of mice, having appetizing effect and improving gastric emptying and the movement of gastrointestinal tract. However, Apazin^® did not show obvious effect on the mice’s weight.

 

 

Materials:

 

1. Apazin^® extract (0.13g equivalent to 1g crude drug) was provided by LIFEnhance, Inc., lot number 20020401 and stored at 4Cº until use.  Extract was dissolved in distilled water to required concentrations.

 

2. Janwei Xiaoshi Pian (a digestive medicine), was provided by Jianzhong Medicine Industry Joint Stock Co. Ltd, lot number 0203064.

Analytically pure by 721 spectrophotometer analysis.

 

3. Animals were obtained from Hubei Animal Experimental center.  Kunming mice weighed 18-20 ± 1g and equal numbers of ♀ (male) and ♂ (female) mice were used.  The certificate of inspection reference number is 10-082.  The mice feed was purchased from Hubei Animal Experimental Center feed factory.

 

Methods and Results:

 

1.      Effect on Appetite

 

Consistent with previously published protocols^[1], 60 mice equal numbers ♀♂ weight 19±1g were divided into five groups at random, the control, Jianwei Xiaoshi Pian group, and three Apazin^® dosage groups. The mice were fed (ig) saline (20ml/kg), 2.4g of Jianwei Xiaoshi Pian, or 5g, 2.5g 1.25g of Apazin^® in saline (20ml/kg) daily for 14 days.  Mice from different groups were raised in different cages.  Feed consumed was determined daily by weighing dried feed before placement into cages followed by weighing unconsumed feed.  Feed consumption and weights of control and test groups were recorded and compared. The statistical significance analysis was determined and listed below in Table1 and Table2.

 

        Table1 Effect on Mice Feed Consumption (g X±SD)

1         21.68±7.03  21.84±7.74   20.31±5.66     21.46±1.55    21.80±4.73

2   24.57±5.08   29.44±5.19^*  25.64±8.49     25.25±1.64    30.51±4.47^**

3   29.91±7.18   28.41±6.32   26.08±5.89     27.69±2.60    31.45±5.64

4   33.34±6.53   31.72±7.25   32.78±8.48     30.89±3.78    36.86±9.44

5   32.75±10.97  29.32±7.79   30.99±5.80     30.61±2.40    33.43±11.51

6   30.44±6.14   29.82±8.72   29.92±8.43     31.90±2.36    33.53±6.48

7   29.27±7.86   28.38±5.99   29.93±5.01     31.34±4.39    35.30±12.63

8   24.85±5.62   26.83±8.19   30.66±5.69^*    28.66±5.47    37.48±11.03^**

9   22.77±3.70   27.88±3.05^*   32.37±7.70^***   28.01±7.90^*   37.73±14.41^*

10  24.52±6.50   26.92±4.35   33.81±10.16^*   32.02±6.96^*   34.84±8.83^**

11  26.32±1.42   28.03±6.92   37.34±14.45^*   33.45±8.35    36.73±12.83^*

12  29.30±3.19   31.49±8.75   36.16±9.29^*    35.62±7.18^*   37.58±13.10^*

13  26.97±4.35   31.32±3.24^*   34.89±8.72^*    34.50±8.40^*   33.02±9.75^*

_____________________________________________________________________

n=12

Compared with the control group *p<0.05   **p<0.01   ***p<0.001

 1   19.38±1.27   19.5±1.22   19.43±1.00  19.12±0.97    19.65±1.00

 2   21.39±1.05   21.04±1.52  21.04±1.12  20.37±1.11    21.57±1.47

 3   22.04±1.19   22.17±1.80  22.31±1.30  21.88±1.09    22.85±1.48

 4   23.39±1.20   23.23±2.23  23.43±1.29  23.07±1.41    23.94±1.52

 5   23.97±1.29   23.63±2.34  24.61±1.28  23.61±1.17    24.46±1.94

 6   25.08±1.77   25.57±1.70  25.41±1.58  24.60±1.48    25.45±1.77

 7   25.90±1.91   25.63±2.80  26.31±1.98  25.68±2.03    26.39±2.12

 8   27.18±2.34   26.72±3.05  27.42±2.24  26.77±2.27    27.58±2.38

 9   28.15±2.66   27.44±3.41  28.27±2.61  27.28±3.23    28.05±2.47

10   27.88±3.31   27.84±3.49  28.80±2.75  27.80±3.22   28.29±2.44

11   28.69±3.70   28.62±3.72  29.37±3.24  28.27±3.30   29.18±3.30

12   28.78±3.81   28.59±3.41  29.47±3.19  28.41±3.27   29.34±3.46

13   29.56±4.18   29.13±3.43  30.23±3.35  29.17±3.65   29.97±3.46

 

Both the Jianwei Xiaoshi Pian group and the three Apazin^® groups demonstrate increased feed consumption.  Despite increased consumption levels there is no significant change in weight.

 

2 .Effect on the Small Intestine Movement

   (Activated char powder method)

In accordance with published protocols ^[2] mice were divided as above. After 12 consecutive daily (ig) Apazin^® extract doses, mice were starved for 24 hours.  To determine small intestinal movement, activated char powder was suspended to 10℅ final concentration in saline solution, Jianwei Xiaoshi Pian, or Apazin then administered (ig) at 0.2ml/10g weight. After 20 minutes, the mice were sacrificed by dislocation of cervical vertebra and the stomach and intestines removed.  Stomach and intestines are placed on a glass board to measure the distance of activated char powder movement and small intestine (from pylorus to ileocecum) total length.  The percentage of movement was calculated as follows:

 

Moving Percentage of activated char powder（%）=

 

   The distance from the front of char powder to pylorus (cm)

  ---------------------------------------------------------------  ×  100%

          the whole length of small intestine(cm）

 

The statistical significances determined are listed in Table 3.  

Groups    Dosage    Total small       dist char powder      Percentage

                     Intestine length       moved           movement

The control     --       41.19±5.48         17.37±3.47          42.56±9.27

Jianwei        2.4      40.58±3.13         20.60±2.56^*         51.19±8.26^*

Large dosage  5.0      39.87±5.9          20.08±2.17^*          50.92±6.83^*   

Medium dosage 2.5      38.15±4.0          20.61±3.68^*         53.87±7.50^**  

n=12

Compared with the control *P<0.05    *P<0.01

  

 

5g and 2.5g doses of Apazin^® can significantly increase the migration distance and the percentage of movement for the char powder indicator in comparison with control.  Apazin^® increases gastrointestinal peristalsis thereby aiding in digestion and improving appetite.

 

3. Quantitative Measurements with Phenol Red

According to established protocols[3], 50 Kunming mice with equal numbers of ♀(male) or ♂(female) were divided into five groups, and fed daily for 12 days (ig) saline, or Jianwei Xiaoshi Pian (2.4g/kg), 6g, 3g, 1.5g/kg of Apazin™ dissolved in saline (0.02ml/g wt).  The mice were then starved for 36 hours followed by 0.01% phenol red administration (i.g) at 0.02ml/g wt.  Mice were sacrificed 20min later to remove small intestinal contents.  The optical density (OD) of contents was measured at 560 nm wavelength using a 721 spectrophotometer. Readings were normalized to phenol red OD standard curve. (See Table 4)

 

 Table 4 Effect on Phenol red Gastrointestinal Excretion

The control        ----          10           14.55±4.25 

Jianwei           2.4          10           18.70±3.45^*      <0.05      

Large dosage      6           10           21.62±4.68^**      <0.01  

Medium dosage    3.0         10           19.96±4.69^*       <0.05  

Results indicate a significant increase in gastrointestinal excretion of phenol red at 6g and 3g doses in addition to greater phenol red retention compared with the control group.

 

References:

[1][3]Li Yikui .Experimental Methodology of Pharmacology of Traditional Chinese Medicine [m] Shanghai: Shanghai Science and Technology Press, 1999:149,321

[2]Chen Qi. Research Methodology of Pharmacology of Traditional Chinese Medicine [M] Beijing: People’s Health Press, 1993:335