Traditional Chinese medicine compositions for alleviating stress

Abstract

The present invention relates to the technical field of health-care product, and in particular relates to a traditional Chinese medicine composition having a function of regulating emotions, a process for preparing the same, and a product thereof. It was found that the composition of the present invention had effects of nourishing liver and kidney, regulating Yin and Yang, tonifying Qi and nourishing blood, regulating emotions, and protecting the function of autonomic nerve, and could improve pressure-induced anxiety and depression, inattention, poor spirit, dispirited, learning and memory capacity decline, etc. The invention has the advantages of specificity and remarkable effect, and it uses all natural medicinal and edible plants as raw materials, the combination of which is simple, safe and stable, and is suitable for long-term use.

Claims

1. A traditional Chinese medicine composition in a dose effective for alleviating stress, the composition made from raw materials consisting of, in specified parts by weight: 40-60 parts of ginseng, 40-60 parts of Radix paeoniae Alba, 40-60 parts of Tree Peony Bark, 40-60 parts of Sharpleaf Galangal Fruit, and 40-60 parts of Radix Polygonum multiflorum Preparata.

2. The traditional Chinese medicine composition according to claim 1, wherein the raw materials consist of, in specified parts by weight: 60 parts of ginseng, 60 parts of Radix paeoniae Alba, 60 parts of Tree Peony Bark, 45 parts of Sharpleaf Galangal Fruit, and 45 parts of Radix Polygonum multiflorum Preparata.

3. A method of alleviating stress in a subject, comprising administrating an effective amount of the composition of claim 1 to a subject in need thereof.

Description

EMBODIMENTS

(1) The present invention discloses a traditional Chinese medicine composition having a function of regulating emotions, a process for preparing the same, and a product thereof, which may be implemented with suitable modifications of the process parameters by those skilled in the art in light of the present disclosure. All the similar alterations and modifications are clear to those skilled in the art and deemed to be included in the present invention. Methods and uses of the present invention have been described by the preferred examples, and it is obvious that those in related art are able to make changes or appropriate alternations and the combinations thereof to the methods and uses described herein to implement and apply the inventive technology without departing from the disclosure, spirit and scope of the present invention.

(2) The ingredients or auxiliaries used in the traditional Chinese medicine composition having a function of regulating emotions, a process for preparing the same, and a product thereof in the present invention are all commercially available.

(3) The raw materials used in the above examples including ginseng, Radix paeoniae Alba, Tree Peony Bark, Sharpleaf Galangal Fruit, and Radix Polygonum multiflorum Preparata in accordance with the quality standard of Chinese Pharmacopoeia (2015).

(4) The present invention is further explained in combination with the examples below.

Example 1

(5) The traditional Chinese medicine composition having a function of regulating emotions provided by the present example was made from the following herbal materials in specified portions by weight: 60 portions of ginseng, 60 portions of Radix paeoniae Alba, 60 portions of Tree Peony Bark, 45 portions of Sharpleaf Galangal Fruit, and 45 portions of Radix Polygonum multiflorum Preparata.

(6) The preparation method comprised the following steps: weighting ginseng 60 g, Radix paeoniae Alba 60 g, Tree Peony Bark 60 g, Sharpleaf Galangal Fruit 45 g, and Radix Polygonum multiflorum Preparata 45 g, conducting decoction with 10 folds of boiling water for 2 hours, extracting twice and combining the filtrate, concentrating to 300 mL.

Experimental Example 1

Pharmacodynamic Studies

(7) In order to confirm the efficacy of the Chinese herbal medicine composition provided by Example 1 of the present invention, the inventor prepared the water extract by the conventional process using the raw material ratio provided in the above Example 1, and conducted the efficacy test. The results were as follows:

(8) 1. Experimental Purpose

(9) To investigate the effect of the Chinese herbal medicine extract on relieving anxiety and depressive symptoms and protecting the function of autonomic nerve.

(10) 2. Experimental Materials

(11) 2.1 Experimental Sample

(12) According to the raw material ratio provided in Example 1, weighting ginseng 60 g, Radix paeoniae Alba 60 g, Tree Peony Bark 60 g, Sharpleaf Galangal Fruit 45 g, and Radix Polygonum multiflorum Preparata 45 g, conducting decoction with 10 folds of boiling water for 2 hours, extracting twice and combining the filtrate, concentrating to 300 mL.

(13) The preparation method of the extract of each raw material including ginseng, Radix paeoniae Alba, Tree Peony Bark, Sharpleaf Galangal Fruit, and Radix Polygonum multiflorum Preparata was the same as that of the composition of Example 1.

(14) 2.2 Modeling and Administration

(15) The rats were housed in individual cages with light and temperature control (temperature: 212 C.; lights on at 8:00 AM and off at 8:00 PM). Food and water were available ad libitum. All experimental procedures were carried out in strict accordance with the National Institutes of Health guidelines for care and use of experimental animals. After an initial habituation period of 1 week, all animals were given access to bottle containing a sucrose solution in tap water for 1 hour and were tested for the baseline data of the sucrose preference test. According to the baseline data, the animals were divided into 9 groups (n=10-12 animals/each group), that is the control group, the stressed group, the stressed+fluoxetine group, the stressed+Example 1 group, the stressed+ginseng extract group, the stressed+Radix paeoniae Alba extract group, the stressed+Tree Peony Bark extract group, the stressed+Sharpleaf Galangal Fruit extract group, and the stressed+Radix Polygonum multiflorum Preparata extract group. The animals were subjected to 2 to 5 different mild stressors per day for 5 weeks. The mild stressors included water and food deprivation, cage tilt, continuously lighting, cage soiling, stroboscopic lighting, restricted access to food, and tail pinching. Control animals were not subjected to the stress protocol, and the stressed animals were subjected to the mild stressors and given the same amount of water as control group. The animals of the stressed+fluoxetine group were subjected to the mild stressors and treated with fluoxetine capsules. The animals of the stressed+Example 1 group were subjected to the mild stressors and treated with the composition of Example 1. The animals of the stressed+single extract group were subjected to the mild stressors and treated with the single extract.

(16) 2.3 Sucrose Preference Test

(17) The rats could select between two bottles of liquid freely, one with 0.8% sucrose solution and one with tap water, after 23 h food and water deprivation. To avoid the rats prefer to drink one side of the water, the position of the two bottles were exchanged every half an hour. Tap water consumption and sucrose consumption were measured by weighting the bottles, and the sucrose preference were calculated based on the following formula:

(18) $\mathrm{SP}=\frac{\mathrm{sucrose}\mathrm{intake}\left(g\right)}{\mathrm{sucrose}\mathrm{intake}\left(g\right)+\mathrm{water}\mathrm{intake}\left(g\right)}100\%$

(19) 2.4 Neurotransmitter Measurement

(20) The rats were sacrificed and the trunk blood was collected into the collection tubes coated with standard heparin. The collection tubes were centrifuged in 4 C. centrifugal machine at 3000 rpm for 15 min. The supernatant was collected and stored at 20 C. until analysis. The levels of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and cortisol (CORT) were detected by ELISA.

(21) 3. Experimental Results

(22) 3.1 Results of Sucrose Preference Test

(23) According to compare the sucrose preference after the stress experiment of each group (as shown in Table 1), it was found that fluoxetine and Example 1 could significantly increase the sucrose preference compared with the stressed group, and it indicated that Example 1 had the effect of improving the symptoms of anhedonia in the chronically stressed rats, and the efficacy was superior to the single herb extract.

(24) TABLE-US-00001 TABLE 1 The effect of Example 1 on the sucrose preference of chronically stressed rats Number of Group Animals Sucrose Preferenc control group 11 0.7694 0.0295 stressed group 10 0.6054 0.0633* stressed + fluoxetine group 12 0.7431 0.0565.sup.# stressed + Example 1 group 12 0.7384 0.0702.sup.# stressed + ginseng extract 12 0.6534 0.0740.sup.# group stressed + Radix paeoniae 12 0.6101 0.0423 Alba extract group stressed + Tree Peony Bark 12 0.6043 0.0735 extract group stressed + Sharpleaf 12 0.6632 0.0242# Galangal Fruit extract group stressed + Radix 12 0.6078 0.0345 Polygonum multiflorum Preparata extract group

(25) 3.2 Results of Neurotransmitter Measurement

(26) As shown in Table 2, the results of neurotransmitter measurement were indicated that chronic stress could significantly increase CRH levels in rats, and significantly reduce ACTH and CORT levels in rats. Fluoxetine and Example 1 could improve the abnormal levels of CRH, ACTH and CORT caused by chronic stress, and it indicated that Example 1 had a certain neuroprotective effect, and the efficacy of it was superior to each of the single herb extract.

(27) TABLE-US-00002 TABLE 2 The effect of Example 1 on the neurotransmitter of chronically stressed rats Number of CRH CORT Group Animals (ng/ml) ACTH (pg/ml) (ng/ml) control group 10 5.01 0.38 34.15 4.27 53.73 5.54 stress group 10 5.97 0.36* 25.45 2.68* 42.34 4.94* stressed + fluoxetine group 10 5.13 0.25.sup.# 33.83 2.26.sup.# 52.09 5.95.sup.# stressed + Example 1 10 5.16 0.29.sup.# 33.28 3.75.sup.# 54.08 1.84.sup.# group stressed + ginseng extract 12 5.82 0.33 26.34 3.45 50.01 3.34.sup.# group stressed + Radix paeoniae 12 5.69 0.58 28.23 1.89 43.22 5.34 Alba extract group stressed + Tree Peony 12 5.76 0.66 27.67 2.31 45.15 5.01 Bark extract group stressed + Sharpleaf 12 5.61 0.40 30.03 2.53.sup.# 51.23 3.32.sup.# Galangal Fruit extract group stressed + Radix 12 5.65 0.31 29.34 5.43.sup.# 46.04 2.35 Polygonum multiflorum Preparata extract group Notes: *presents a comparison to control group, p < 0.05; .sup.#presents a comparison to model group, p < 0.05.

(28) 4. Summary of Efficacy

(29) As shown in the above experiments, Example 1 could improve anhedonia in chronically stressed rat model and the abnormal levels of the neurotransmitter, and it had a significant effect of modulating mood and protecting the function of nervous system. Meanwhile, the experimental results indicated that the combination of ginseng, Radix paeoniae Alba, Tree Peony Bark, Sharpleaf Galangal Fruit, and Radix Polygonum multiflorum Preparata had a certain synergistic effect.

Example 2

(30) The traditional Chinese medicine composition having a function of regulating emotions provided by the present invention was made from the following herbal materials in specified portions by weight: 80 portions of ginseng, 80 portions of Radix paeoniae Alba, 80 portions of Tree Peony Bark, 20 portions of Sharpleaf Galangal Fruit, and 20 portions of Radix Polygonum multiflorum Preparata.

(31) The preparation method was the same as that of Example 1.

Example 3

(32) The traditional Chinese medicine composition having a function of regulating emotions provided by the present invention was made from the following herbal materials in specified portions by weight: 50 portions of ginseng, 50 portions of Radix paeoniae Alba, 50 portions of Tree Peony Bark, 50 portions of Sharpleaf Galangal Fruit, and 50 portions of Radix Polygonum multiflorum Preparata.

(33) The preparation method was the same as that of Example 1.

Example 4

(34) The traditional Chinese medicine composition having a function of regulating emotions provided by the present invention was made from the following herbal materials in specified portions by weight: 40 portions of ginseng, 40 portions of Radix paeoniae Alba, 40 portions of Tree Peony Bark, 60 portions of Sharpleaf Galangal Fruit, and 60 portions of Radix Polygonum multiflorum Preparata.

(35) The preparation method was the same as that of Example 1.

Example 5

(36) The traditional Chinese medicine composition having a function of regulating emotions provided by the present invention was made from the following herbal materials in specified portions by weight: 60 portions of ginseng, 60 portions of Radix paeoniae Alba, 60 portions of Tree Peony Bark, 40 portions of Sharpleaf Galangal Fruit, and 60 portions of Radix Polygonum multiflorum Preparata.

(37) The preparation method was the same as that of Example 1.

Example 6

(38) The traditional Chinese medicine composition having a function of regulating emotions provided by the present invention was made from the following herbal materials in specified portions by weight: 20 portions of ginseng, 80 portions of Radix paeoniae Alba, 20 portions of Tree Peony Bark, 80 portions of Sharpleaf Galangal Fruit, and 20 portions of Radix Polygonum multiflorum Preparata.

(39) The preparation method was the same as that of Example 1.

Example 7

(40) The traditional Chinese medicine composition having a function of regulating emotions provided by the present invention was made from the following herbal materials in specified portions by weight: 40 portions of ginseng, 60 portions of Radix paeoniae Alba, 40 portions of Tree Peony Bark, 60 portions of Sharpleaf Galangal Fruit, and 40 portions of Radix Polygonum multiflorum Preparata.

(41) The preparation method was the same as that of Example 1.

Experimental Example 2

Pharmacodynamic Studies

(42) In order to confirm the efficacy of the Chinese herbal medicine composition provided by Examples 2-7 of the present invention, the inventor prepared the water extract by the conventional process using the raw material ratio provided in the above Example 2-7, and conducted the efficacy test. The results were as follows:

(43) 1. Experimental Purpose

(44) To investigate the effect of the Chinese herbal medicine extract on relieving anxiety and depressive symptoms and protecting the function of autonomic nerve.

(45) 2. Experimental Materials

(46) 2.1 Experimental Sample

(47) The extract of Examples 2-7.

(48) 2.2 Modeling and Administration

(49) The rats were housed in individual cages with light and temperature control (temperature: 212 C.; lights on at 8:00 AM and off at 8:00 PM). Food and water were available ad libitum. All experimental procedures were carried out in strict accordance with the National Institutes of Health guidelines for care and use of experimental animals. After an initial habituation period of 1 week, all animals were given access to bottle containing a sucrose solution in tap water for 1 hour and were tested for the baseline data of the sucrose preference test. According to the baseline data, the animals were divided into 9 groups (n=10-12 animals/each group), that is the control group, the stressed group, the stressed+fluoxetine group, the stressed+Example 2 group, the stressed+Example 3 group, the stressed+Example 4 group, the stressed+Example 5 group, the stressed+Example 6 group, and the stressed+Example 7 group. The animals were subjected to 2 to 5 different mild stressors per day for 5 weeks. The mild stressors included water and food deprivation, cage tilt, continuously lighting, cage soiling, stroboscopic lighting, restricted access to food, and tail pinching. Control animals were not subjected to the stressed protocol, and stressed animals were subjected to the mild stressors and given the same amount of water as control group. The animals of the stressed+fluoxetine group were subjected to the mild stressors and treated with fluoxetine capsules. The animals of the stressed+Example 2-7 groups were subjected to the mild stressors and treated with the compositions of Example 2-7.

(50) 2.3 Sucrose Preference Test

(51) The rats could select between two bottles of liquid freely, one with 0.8% sucrose solution and one with tap water, after 23 h food and water deprivation. To avoid the rats prefer to drink one side of the water, the position of the two bottles were exchanged every half an hour. Tap water consumption and sucrose consumption were measured by weighting the bottles, and the sucrose preference were calculated based on the following formula:

(52) $\mathrm{SP}=\frac{\mathrm{sucrose}\mathrm{intake}\left(g\right)}{\mathrm{sucrose}\mathrm{intake}\left(g\right)+\mathrm{water}\mathrm{intake}\left(g\right)}100\%$

(53) 2.4 Neurotransmitter Measurement

(54) The rats were sacrificed and the trunk blood was collected into the collection tubes coated with standard heparin. The collection tubes were centrifuged in 4 C. centrifugal machine at 3000 rpm for 15 min. The supernatant was collected and stored at 20 C. until analysis. The levels of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and cortisol (CORT) were detected by ELISA.

(55) 3. Experimental Results

(56) 3.1 Results of Sucrose Preference Test

(57) According to compare the sucrose preference after the stress experiment of each group (as shown in Table 3), it was found that fluoxetine and Examples 2-7 could significantly increase the sucrose preference to varying degrees compared with the stressed group, and it indicated that Example 2-7 had the effect of improving the symptoms of anhedonia in the chronically stressed rats.

(58) TABLE-US-00003 TABLE 3 The effects of Example 2-7 on the sucrose preference of chronically stressed rats Number of Group Animals Sucrose Preference control group 11 0.7694 0.0295 stressed group 10 0.6054 0.0633* stressed + fluoxetine 12 0.7431 0.0565.sup.# group stressed + Example 2 12 0.6462 0.0624 group stressed + Example 3 12 0.6962 0.0534.sup.# group stressed + Example 4 12 0.6845 0.0712.sup.# group stressed + Example 5 12 0.7001 0.0213.sup.# group stressed + Example 6 12 0.6221 0.0431 group stressed + Example 7 12 0.7003 0.0421.sup.# group

(59) 3.2 Results of Neurotransmitter Measurement

(60) As shown in Table 4, the results of neurotransmitter measurement were indicated that chronic stress could significantly increase CRH levels in rats, and significantly reduce ACTH and CORT levels in rats. Fluoxetine and Example 2-7 could improve the abnormal levels of CRH, ACTH and CORT caused by chronic stress, and it indicated that Example 2-7 had a certain neuroprotective effect.

(61) TABLE-US-00004 TABLE 4 The effects of Example 2-7 on the neurotransmitter of chronically stressed rats Number of Group Animals CRH (ng/ml) ACTH (pg/ml) CORT (ng/ml) control group 10 5.01 0.38 34.15 4.27 53.73 5.54 stressed group 10 5.97 0.36* 25.45 2.68* 42.34 4.94* stressed + 10 5.13 0.25.sup.# 33.83 2.26.sup.# 52.09 5.95.sup.# fluoxetine group stressed + 12 5.18 0.24.sup.# 28.33 1.98 44.01 2.29 Example 2 group stressed + 12 5.53 0.45 34.34 2.21.sup.# 51.23 4.03.sup.# Example 3 group stressed + 12 5.12 0.24.sup.# 26.12 3.45 50.03 4.13.sup.# Example 4 group stressed + 12 5.78 0.29 30.13 5.12.sup.# 54.23 3.21.sup.# Example 5 group stressed + 12 5.59 0.39 29.12 4.23.sup.# 43.24 3.56 Example 6 group stress + 12 5.07 0.21.sup.# 24.36 4.01 51.98 6.39.sup.# Example 7 group Notes: *presents a comparison to control group, p < 0.05; .sup.#presents a comparison to model group, p < 0.05.

(62) 4. Summary of Efficacy

(63) As shown in the above experiments, Examples 2-7 could improve anhedonia in chronic stress rat model and the abnormal levels of the neurotransmitter, and it had an effect of modulating mood and protecting the function of nervous system. Meanwhile, the experimental results indicated that the combination of ginseng, Radix paeoniae Alba, Tree Peony Bark, Sharpleaf Galangal Fruit, and Radix Polygonum multiflorum Preparata had a certain synergistic effect.

Comparative Example 1

(64) The traditional Chinese medicine composition having a function of regulating emotions provided by the present comparative example was made from the following herbal materials in specified portions by weight: 10 portions of ginseng, 120 portions of Radix paeoniae Alba, 10 portions of Tree Peony Bark, 120 portions of Sharpleaf Galangal Fruit, and 10 portions of Radix Polygonum multiflorum Preparata.

(65) The preparation method was the same as that of Example 1.

Comparative Example 2

(66) The traditional Chinese medicine composition having a function of regulating emotions provided by the present comparative example was made from the following herbal materials in specified portions by weight: 120 portions of ginseng, 10 portions of Radix paeoniae Alba, 120 portions of Tree Peony Bark, 10 portions of Sharpleaf Galangal Fruit, and 120 portions of Radix Polygonum multiflorum Preparata.

(67) The preparation method was the same as that of Example 1.

Comparative Example 3

(68) The traditional Chinese medicine composition having a function of regulating emotions provided by the present comparative example was made from the following herbal materials in specified portions by weight: 60 portions of Radix paeoniae Alba, 60 portions of Tree Peony Bark, 45 portions of Sharpleaf Galangal Fruit, and 45 portions of Radix Polygonum multiflorum Preparata.

(69) The preparation method was the same as that of Example 1.

Comparative Example 4

(70) The traditional Chinese medicine composition having a function of regulating emotions provided by the present comparative example was made from the following herbal materials in specified portions by weight: 60 portions of ginseng, 60 portions of Tree Peony Bark, 45 portions of Sharpleaf Galangal Fruit, and 45 portions of Radix Polygonum multiflorum Preparata.

(71) The preparation method was the same as that of Example 1.

Comparative Example 5

(72) The traditional Chinese medicine composition having a function of regulating emotions provided by the present comparative example was made from the following herbal materials in specified portions by weight: 60 portions of ginseng, 60 portions of Radix paeoniae Alba, 45 portions of Sharpleaf Galangal Fruit, and 45 portions of Radix Polygonum multiflorum Preparata.

(73) The preparation method was the same as that of Example 1.

Comparative Example 6

(74) The traditional Chinese medicine composition having a function of regulating emotions provided by the present comparative example was made from the following herbal materials in specified portions by weight: 60 portions of ginseng, 60 portions of Radix paeoniae Alba, 60 portions of Tree Peony Bark, and 45 portions of Radix Polygonum multiflorum Preparata.

(75) The preparation method was the same as that of Example 1.

Comparative Example 7

(76) The traditional Chinese medicine composition having a function of regulating emotions provided by the present comparative example was made from the following herbal materials in specified portions by weight: 60 portions of ginseng, 60 portions of Radix paeoniae Alba, 60 portions of Tree Peony Bark, and 45 portions of Sharpleaf Galangal Fruit.

(77) The preparation method was the same as that of Example 1.

Comparative Example 1

Pharmacodynamic Studies

(78) In order to confirm the efficacy of the Chinese herbal medicine composition provided by Comparative Example 1-7 of the present invention, the inventor prepared the water extract by the conventional process using the raw material ratio provided in the above Comparative Example 1-7, and conducted the efficacy test. The results were as follows:

(79) 1. Experimental Purpose

(80) To investigate the effect of the Chinese herbal medicine extract on relieving anxiety and depressive symptoms and protecting the function of autonomic nerve.

(81) 2. Experimental Materials

(82) 2.1 Experimental Sample

(83) The extract of Comparative Example 1-7.

(84) 2.2 Modeling and Administration

(85) The rats were housed in individual cages with light and temperature control (temperature: 212 C.; lights on at 8:00 AM and off at 8:00 PM). Food and water were available ad libitum. All experimental procedures were carried out in strict accordance with the National Institutes of Health guidelines for care and use of experimental animals. After an initial habituation period of 1 week, all animals were given access to bottle containing a sucrose solution in tap water for 1 hour and were tested for the baseline data of the sucrose preference test. According to the baseline data, the animals were divided into 10 groups (n=10-12 each), that is the control group, the stressed group, the stressed+fluoxetine group, the stressed+Comparative Example 1 group, the stressed+Comparative Example 2 group, the stressed+Comparative Example 3 group, the stressed+Comparative Example 4 group, the stressed+Comparative example 5 group, the stressed+Comparative Example 6 group, and the stressed+Comparative Example 7 group. The animals were subjected to 2 to 5 different mild stressors per day for 5 weeks. The mild stressors included water and food deprivation, cage tilt, continuously lighting, cage soiling, stroboscopic lighting, restricted access to food, and tail pinching. Control animals were not subjected to the stress protocol, and stressed animals were subjected to the mild stressors and given the same amount of water as control group. The animals of the stressed+fluoxetine group were subjected to the mild stressors and treated with fluoxetine capsules. The animals of the stressed+Comparative Example 1-7 groups were subjected to the mild stressors and treated with the compositions of Comparative Example 1-7.

(86) 2.3 Sucrose Preference Test

(87) The rats could select between two bottles of liquid freely, one with 0.8% sucrose solution and one with tap water after 23 h food and water deprivation. To avoid the rats prefer to drink one side of the water, the position of the two bottles were exchanged every half an hour. Tap water consumption and sucrose consumption were measured by weighting the bottles, and the sucrose preference were calculated based on the following formula:

(88) $\mathrm{SP}=\frac{\mathrm{sucrose}\mathrm{intake}\left(g\right)}{\mathrm{sucrose}\mathrm{intake}\left(g\right)+\mathrm{water}\mathrm{intake}\left(g\right)}100\%$

(89) 2.4 Neurotransmitter Measurement

(90) The rats were sacrificed and the trunk blood was collected into the collection tubes coated with standard heparin. The collection tubes were centrifuged in 4 C. centrifugal machine at 3000 rpm for 15 min. The supernatant was collected and stored at 20 C. until analysis. The levels of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and cortisol (CORT) were detected by ELISA.

(91) 3. Experimental Results

(92) 3.1 Results of Sucrose Preference Test

(93) According to compare the sucrose preference after the stress experiment of each group (as shown in Table 5), it was found that fluoxetine could significantly increase the sucrose preference compared with the stress group, and it had the effect of improving the symptoms of anhedonia, but comparing with stress group, the sucrose preference of Comparative Example 1-7 were not changed significantly.

(94) TABLE-US-00005 TABLE 5 The effects of Comparative Example 1-7 on the sucrose preference of chronically stressed rats Number of Group Animals Sucrose Preference control group 11 0.8204 0.0195 stress group 10 0.5901 0.0433* stress + fluoxetine group 12 0.7502 0.0471.sup.# stress + Comparative 12 0.6134 0.0453 Example 1 group stress + Comparative 12 0.6212 0.0212 Example 2 group stress + Comparative 12 0.5801 0.0534 Example 3 group stress + Comparative 12 0.6121 0.0321 Example 4 group stress + Comparative 12 0.6019 0.0521 Example 5 group stress + Comparative 12 0.6047 0.0529 Example 6 group stress + Comparative 12 0.6081 0.0487 Example 7 group

(95) 3.2 Results of Neurotransmitter Measurement

(96) As shown in Table 6, the results of neurotransmitter measurement were indicated that chronic stress could significantly increase CRH levels in rats, and significantly reduce ACTH and CORT levels in rats. Fluoxetine rather than Comparative Examples 1-7 could improve the abnormal levels of CRH, ACTH and CORT caused by chronic stress.

(97) TABLE-US-00006 TABLE 6 The effects of Comparative Examples 1-7 on the neurotransmitter of chronically stressed rats Number of CORT Group Animals CRH (ng/ml) ACTH (pg/ml) (ng/ml) control group 10 5.03 0.41 33.17 3.26 54.73 4.84 stressed group 10 5.88 0.25* 26.19 2.68* 41.40 3.98* stressed + fluoxetine 10 5.11 0.29.sup.# 32.73 2.66.sup.# 52.46 6.05.sup.# group stressed + Comparative 12 5.86 0.56 24.76 2.80 43.39 3.45 Example 1 group stressed + Comparative 12 5.90 0.21 25.65 1.98 44.48 3.57 Example 2 group stressed + Comparative 12 6.01 0.22 27.01 1.02 42.33 3.01 Example 3 group stressed + Comparative 12 5.98 0.29 25.10 2.98 42.88 2.10 Example 4 group stressed + Comparative 12 5.84 0.24 25.39 3.11 43.29 1.99 Example 5 group stressed + Comparative 12 5.93 0.38 26.18 2.01 41.90 3.42 Example 6 group stresse + Comparative 12 5.66 0.78 26.16 2.81 43.39 2.01 Example 7 group Notes: *presents a comparison to control group, p < 0.05; .sup.#presents a comparison to model group, p < 0.05.

(98) The above description gives only the preferred embodiments of the present invention, and it should be noted that for those of ordinary skill in the art, a number of improvements and modifications can be made without departing from the principle of the invention, which are also regarded as falling into the scope claimed in the present invention.