According the results observed under the electron microscope, the most severe injuries were observed in model group. This indicated that the model is constructed successfully. On the other hand, much less severe damage was observed in the three saponins family of
Radix notoginshen intravenous injection groups and the positive control group. Furthermore, the cell damage decreased while the amount of saponins family of
Radix notoginshen intravenous injection increased. This indicated that the saponins family of
Radix notoginshen intravenous injection could protect animal from acute incomplete ischemia reperfusion injury.
[TABLE-US-00006]
TABLE 1 |
|
|
Effect of saponins family of Radix notoginshen intravenous injection on |
experimental rat ischemia reperfusion injury (n = 20) |
|
|
|
Brain index |
|
|
Dosage |
EB content (μg/g |
(g brain/100 g |
Brain water |
Group |
(mg/Kg) |
brain wet weight) |
body weight) |
content (%) |
|
Model |
? ? ? |
12.10 ± 0.56 |
0.64 ± 0.02 |
81.17 ± 0.60 |
Pseudo-operation |
? ? ? |
6.81 ± 0.75*** |
0.55 ± 0.02*** |
77.14 ± 1.00*** |
saponins family of Radix |
1940 |
10.56 ± 0.49*** |
0.64 ± 0.01 |
80.09 ± 0.73*** |
notoginshen |
Low dosage of saponins family of |
34.03 |
9.81 ± 0.39*** |
0.63 ± 0.01* |
80.44 ± 0.60** |
Radix notoginshen |
Medium dosage of saponins |
68.06 |
9.61 ± 0.32*** |
0.60 ± 0.01*** |
78.63 ± 0.57*** |
family of Radix notoginshen |
High dosage of saponins family of |
132.12 |
9.55 ± 0.31*** |
0.60 ± 0.01*** |
78.99 ± 0.73*** |
Radix notoginshen |
|
*compared to model group p < 0.05,
|
**p < 0.01,
|
***p < 0.001
|
|
The above results prove that the saponins family of
Radix notoginshen intravenous injection has certain protective effects on experimental rat acute incomplete ischemia reperfusion injury. It is worthy to have further application on clinical practices.
Experiment 7, the Effect of the Saponins Family of
Radix Notoginshen Intravenous Injection on the Cerebrovascular of Anaesthetized Dog
(1) Experimental medicament and its dosage: 5% saponins family of
Radix notoginshen intravenous injection preparation (manufactured by Lizhu Group Limin Pharmaceutical Industry, packed as 100 ml, batch No. 20000301).). The total amount of saponins of
Radix notoginshen intravenous injection preparation used in clinical practice is 350 mg/250 ml per treatment. It could be applied 1˜2 times per day for a 28 day treatment phase. In the experiment, the dosage of the medicament for the experimental animal was calculated according to that for a human by using the transform factor mg/Kg-mg/m
2. A single dose of the equivalent amount of medicament for animals was applied to the dog low dosage group. The medium dosage group received twice the equivalent medicament and high dosage received 4 times the equivalent medicament of the low dosage group. The dosage of saponins family of
Radix notoginshen intravenous injection for an adult is 250 ml once per day. That is, 350 mg of saponins family of
Radix notoginshen is applied per day. In the case of dog, 1) the equivalent amount of medicament is 350 mg/60 kg×35/19?10.75 mg saponins family of
Radix notoginshen 1 kg, 2) low dosage is 10.75×1=10.75 mg/Kg, 3) medium dosage is 10.75×2=21.5 mg/Kg, 4) high dosage is 10.75×4=43 mg/Kg. 5% of total amount of saponins of
Radix notoginshen intravenous injection and Mailuoning injection were diluted before usage to a needed concentration using physiological saline.
The control medicament was Mailuoning injection (manufactured by Nanjing Jinglin Pharmaceutical Corp., medicine content of 10 g/ml, batch No. 960812). The adult dosage was 10 ml (crude drug of 10 g), twice per day, that is, 20 g/60 kg per day. For the dog, 1) equivalent dosage is 20 g/60 kg×35/19?0.61 g/Kg, 2) dosage of medicament is 0.61×4=2.44 g/Kg. Physiological saline was manufactured by Southern Hospital Pharmacy, batch No. 20000926.
(2) Experimental animals: 30 healthy adult hybrid dogs, each weights 11˜15kg, half male and half female.
(3) Experimental conditions: room temperature 25° C., relative humidity 60%˜70%, and regular pasteurization of experimental equipments.
(4) Equipment: electromagnetic flowmeter, model FM-27, manufactured by Japanese Photoelectric Ltd. The [8] Physiological Recording Instrument of model RM-6000, manufactured by Japanese Photoelectric Ltd and Chengdu Instrument Industry.
(5) Method of medicaments application: intravenous injection with a concentration of 30 ml/Kg.
(6) Experimental control: the negative control group was given same amount of physiological saline while the positive control group was given 4 times of the equivalent amount of the Mailuoning injection. (
7) Experimental data statistics
The experimental data was analyzed by statistical t-testing. The physiological statistics were used for data analysis for comparison among all groups and the negative control group before and after medicament applications.
(8) Experimental methods
The experimental animals were randomly grouped into 5 groups including the negative and positive control groups, and the low dosage, medium dosage and high dosage of saponins family of
Radix notoginshen intravenous injection. The dogs were first anaesthetized via an intravenous injection of 30 mg/Kg 3% pentobarbital sodium, and then fixed on the operation platform. 1 mg/ml of pentobarbital sodium diluted in physiological saline was intravenously infused to maintain the anesthesia. The dog neck was cut for about 10cm to separate the left a.carotis communis, and thread on the same side the a.carotis externa and the a.vertebralis. The No. 4 sensor of the electromagnetic flowmeter was put in the a.carotis communis. The blood flow of the a.carotis communis, calculated by the electromagnetic flowmeter, could be considered as cerebral blood flow. At the same time, the a.fermoralis and v.fermoralis on one side were separated. The a.fermoralis was connected to the pressure transducer and v.fermoralis was connected to the 3-way pipe for medicament application. All four legs were connected to the sensors for the electrocardiogram monitor. Meanwhile, blood pressure of the a.fermoralis, II lead electrocardiogram, and the average blood flow of the a.carotis communis were recorded by the [ 8 ] Physiological Recording Instrument. When the average blood flow, electrocardiogram and blood pressure were indicated stable by the equipment, the data were recorded and the medicaments were applied through intravenous injection. The average amount of medicament application was 30 ml/Kg for all groups. At the same time, the cerebral blood flow, blood pressure and cardiac rhythm were recorded at 0 min, 5 min, 15 min, 30 min and 60 min after medicament application. At the end of the experiment, the skull was opened and the whole brain was removed from above the medulla oblongata. The brain was then weighed. The brain weight of one side was obtained by dividing the resulting data by 2. The cerebral blood flow per minute for each gram of brain tissue was obtained by dividing the recorded cerebral blood flow by the brain weight of one side.
(9) Result
1) The effect of the Saponins Family of
Radix Notoginshen Intravenous Injection on the Cardiac Rhythm of Anaesthetized Dog
The experimental results indicated statistically meaningful fluctuations of the cardiac rhythm in the low dosage group recorded after medicament application 5 min and 15 min, and the medium dosage group recorded after 0 min. There were no statistical differences in other groups and time phases (table 2).
2) The Effect of the Saponins Family of
Radix Notoginshen Intravenous Injection on the Blood Pressure of Anaesthetized Dog