Application of dendrobium huoshanense protocorm in resisting fatigue and invigorating kidney and strengthening yang

Abstract

The invention discloses an application of dendrobium huoshanense protocorm in resisting fatigue, invigorating kidney and strengthening yang. The dendrobium huoshanense protocorm is combined with the herba cistanche, the American ginseng, the raspberry, the dogwood and the fructus lycii, so that the anti-fatigue, kidney-tonifying and yang-strengthening effects of the formula can be remarkably improved. Compared with the single use of cistanche, after the addition of the dendrobium huoshanense protocorm, the exhaustive swimming time of a mouse can be obviously prolonged, the content of urea nitrogen and lactic acid in serum of the mouse is reduced, the content of hepatic glycogen in hepatic tissue is improved, and the content of muscle glycogen in muscular tissue is improved. Compared with a single use of cistanche, the dendrobium huoshanense protocorm can significantly improve the activity of superoxide dismutase in serum of rats with kidney-yang deficiency, reduce the content of malondialdehyde in the serum, improve the content of testosterone in the serum, improve the activity of nitric oxide synthase in penis tissues, and improve the content of nitric oxide and cyclic guanosine monophosphate in the penis tissues after the addition of the dendrobium huoshanense protocorm.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and particularly relates to the application of protocorms of Dendrobium huoshanense in anti-fatigue and kidney-tonifying and yang-strengthening Technical Background

[0002] With the continuous increase of life and work pressure, more and more people are in a sub-healthy state. Sub-health is considered to be the pre-stage of many chronic diseases and is also a reversible stage. Preventing the occurrence and development of various chronic diseases can be achieved by alleviating sub-healthy symptoms. Fatigue and kidney-yang deficiency are the main symptoms of sub-health.

[0003] Fatigue belongs to the category of overwork syndrome in traditional Chinese medicine and is mainly caused by qi and blood deficiency and viscera disharmony. As a non-specific symptom, fatigue exists in many chronic diseases. If fatigue is not recovered in time, it will accumulate and lead to overwork, resulting in chronic fatigue syndrome, including decreased immunity, endocrine disorders, organic lesions of organs, sexual dysfunction, etc.

[0004] Erectile dysfunction is an important symptom of kidney-yang deficiency. Traditional Chinese medicine believes that the kidney dominates the qi of the whole body, the root of a person lies in the kidney, and the driving force of sexuality lies in essence. Kidney-yang deficiency is the main pathogenesis of erectile dysfunction, and Chinese herbal medicines for tonifying the kidney and strengthening yang can be used to treat erectile dysfunction diseases. Modern research shows that NO is an important neurotransmitter in the smooth muscle of the corpus cavernosum of the penis. In penile tissue, nitric oxide synthase (NOS) catalyzes the synthesis of NO from arginine. NO enters the smooth muscle cells of the corpus cavernosum and activates guanylate cyclase, converting GTP into cGMP. As a second messenger, cGMP regulates the ion concentration in cells, causing the smooth muscle of the corpus cavernosum to relax, increasing blood flow, and resulting in penile erection. After cGMP is hydrolyzed by phosphodiesterase type 5, this pathway is interrupted and the erection terminates. Disorders of the NOS / NO / cGMP signaling pathway, including decreased NOS activity and reduced levels of NO and cGMP, are important pathogeneses of erectile dysfunction. Activating the NOS / NO / cGMP signaling pathway is an important way to treat erectile dysfunction.

[0005] Cistanche deserticola, American ginseng, Rubus idaeus, Cornus officinalis, and Lycium barbarum are all Chinese herbal medicines with both medicinal and edible properties for warming the kidney and strengthening yang. Chinese herbal medicines with both medicinal and edible properties can not only exert the therapeutic effects of traditional Chinese herbal medicines but also have the safety characteristics of food, meeting people's pursuit of health. However, the effects of the combination of Cistanche deserticola, American ginseng, Rubus idaeus, Cornus officinalis, and Lycium barbarum in anti-fatigue and kidney-tonifying and yang-strengthening are relatively mild and require a long time to take effect.

[0006] The protocorm-like bodies of Dendrobium huoshanense are embryogenic tissues obtained through plant tissue culture using the shoot tip meristem of Dendrobium huoshanense as the explant. The cells of the protocorm-like bodies of Dendrobium huoshanense not only have the ability to replicate and proliferate but also the ability to differentiate into complete plants, possessing the characteristics of stem cells and being a kind of totipotent stem cells of Dendrobium huoshanense. In 2024, the National Health Commission issued an announcement to include the protocorm-like bodies of Dendrobium huoshanense in new food raw materials. The present invention applies the protocorm-like bodies of Dendrobium huoshanense to the formula of Cistanche deserticola, Panax quinquefolius, Rubus idaeus, Cornus officinalis, and Lycium barbarum, significantly improving the anti-fatigue and kidney-tonifying and yang-strengthening effects of the formula. Summary of the Invention

[0007] The purpose of the present invention is to provide a new application of the protocorm-like bodies of Dendrobium huoshanense, which is applied to improving the anti-fatigue and kidney-tonifying and yang-strengthening effects of the formula of Cistanche deserticola, Panax quinquefolius, Rubus idaeus, Cornus officinalis, and Lycium barbarum. The protocorm-like bodies of Dendrobium huoshanense used in the present invention are undifferentiated protocorm-like bodies of Dendrobium huoshanense, and no effect of improving the anti-fatigue and kidney-tonifying and yang-strengthening effects of the formula has been found in the differentiated Dendrobium huoshanense buds or Dendrobium huoshanense plants.

[0008] The application method of the protocorm-like bodies of Dendrobium huoshanense in anti-fatigue and kidney-tonifying and yang-strengthening includes the following steps:

[0009] 1. Add 3 parts by weight of dry products of the protocorm-like bodies of Dendrobium huoshanense to a formula with 5 parts by weight of Cistanche deserticola, 5 parts by weight of Panax quinquefolius, 3 parts by weight of Rubus idaeus, 2 parts by weight of Cornus officinalis, and 1 part by weight of Lycium barbarum, and superfine pulverize to make powder for standby;

[0010] 2. Add the superfine powder to the gastric juice simulation solution with a pH of 2.0 and a pepsin activity of 2000 U / mL according to a solid-liquid ratio of 1:50, stir at 37°C for 2 h, then adjust the pH to 6.8 with a 1 mol / L NaOH solution to inactivate pepsin. Then add trypsin and pancreatic amylase until the activities are both 100 U / mL, continue to stir at 37°C for 2 h, heat to 95°C and keep for 5 min to inactivate the enzymes, filter to obtain the supernatant, and concentrate and dry the supernatant to obtain a bionic extract;

[0011] 3. The bionic extract can be used in traditional Chinese medicine preparations or functional foods for anti-fatigue and kidney-tonifying and yang-strengthening.

[0012] The beneficial technical effects of the present invention are reflected in that: compared with using the Cistanche deserticola formula alone, adding the protocorm-like bodies of Dendrobium huoshanense can significantly prolong the exhaustive swimming time of mice, reduce the contents of blood urea nitrogen and lactic acid in the serum of mice, increase the glycogen content in the liver tissue, and increase the glycogen content in the muscle tissue; compared with using the Cistanche deserticola formula alone, adding the protocorm-like bodies of Dendrobium huoshanense can significantly increase the superoxide dismutase activity in the serum of kidney-yang deficiency rats, reduce the content of malondialdehyde in the serum, increase the content of testosterone in the serum, increase the nitric oxide synthase activity in the penile tissue, and increase the contents of nitric oxide and cyclic guanosine monophosphate in the penile tissue. Description of the Drawings

[0013] Figure 1 Protocorm-like bodies of Dendrobium huoshanense Specific implementation manners

[0014] The present invention will be further described below in conjunction with specific embodiments. The features and advantages of the present invention will become clearer with the description, but the exemplary embodiments are only used to illustrate the present invention and do not constitute any limitation to the scope of the present invention.

[0015] Example 1

[0016] Preparation of bionic extract of Cistanche deserticola formula: Weigh 5 parts of Cistanche deserticola, 5 parts of American ginseng, 3 parts of Rubus idaeus, 2 parts of Cornus officinalis, and 1 part of Lycium barbarum according to parts by weight, and ultrafinely pulverize them into powder for standby; add the ultrafine powder to the gastric juice simulation solution with a pH of 2.0 and a pepsin activity of 2000 U / mL according to a material-liquid ratio of 1:50, stir at 37 °C for 2 h, then adjust the pH to 6.8 with a 1 mol / L NaOH solution to inactivate pepsin, then add trypsin and pancreatic amylase until the activities are both 100 U / mL, continue to stir at 37 °C for 2 h, then heat to 95 °C and keep for 5 min to inactivate the enzymes, filter to obtain the supernatant, and the supernatant is concentrated and dried to obtain the bionic extract of Cistanche deserticola formula.

[0017] Preparation of Protocorm-like bodies of Dendrobium huoshanense + bionic extract of Cistanche deserticola formula: Weigh 5 parts of Cistanche deserticola, 5 parts of American ginseng, 3 parts of Rubus idaeus, 2 parts of Cornus officinalis, 1 part of Lycium barbarum, and 3 parts of dry Protocorm-like bodies of Dendrobium huoshanense according to parts by weight, and ultrafinely pulverize them into powder for standby; add the ultrafine powder to the gastric juice simulation solution with a pH of 2.0 and a pepsin activity of 2000 U / mL according to a material-liquid ratio of 1:50, stir at 37 °C for 2 h, then adjust the pH to 6.8 with a 1 mol / L NaOH solution to inactivate pepsin, then add trypsin and pancreatic amylase until the activities are both 100 U / mL, continue to stir at 37 °C for 2 h, then heat to 95 °C and keep for 5 min to inactivate the enzymes, filter to obtain the supernatant, and the supernatant is concentrated and dried to obtain Protocorm-like bodies of Dendrobium huoshanense + bionic extract of Cistanche deserticola formula.

[0018] Example 2

[0019] Verification test on the effect of Dendrobium huoshanense protocorms in enhancing the anti-fatigue efficacy of the formula composed of Cistanche deserticola, Panax quinquefolius, Rubus idaeus, Cornus officinalis, and Lycium barbarum. Mice were used as experimental animals and randomly divided into a blank control group (Group 0), a Cistanche deserticola formula group (Group 1), and a Dendrobium huoshanense protocorms + Cistanche deserticola formula group (Group 2). The blank control group was intragastrically administered distilled water, the Cistanche deserticola formula group was intragastrically administered the bionic extract of the Cistanche deserticola formula, and the Dendrobium huoshanense protocorms + Cistanche deserticola formula group was intragastrically administered the bionic extract of Dendrobium huoshanense protocorms + Cistanche deserticola formula. The dosage of each extract was 900 mg / kg, and the intragastric administration volume was 0.2 mL. Each experimental group was administered once a day for 30 consecutive days. On the 30th day after the first administration, the exhaustive swimming time of the mice was measured; the contents of urea nitrogen and lactic acid in the serum were measured; the content of hepatic glycogen in the liver tissue was measured; and the content of muscle glycogen in the muscle tissue was measured.

[0020] The results of the verification test on the effect of Dendrobium huoshanense protocorms in enhancing the anti-fatigue efficacy of the formula composed of Cistanche deserticola, Panax quinquefolius, Rubus idaeus, Cornus officinalis, and Lycium barbarum are shown in Table 1, Table 2, and Table 3.

[0021] Table 1 Results of the exhaustive swimming time test of mice

[0022]

[0023] Note: a indicates significant difference compared with the blank control group (P ≤ 0.05), and b indicates significant difference compared with the Cistanche deserticola formula group (P ≤ 0.05).

[0024] Table 2 Results of the test on the contents of urea nitrogen and lactic acid in the serum of mice

[0025]

[0026] Note: a indicates significant difference compared with the blank control group (P ≤ 0.05), and b indicates significant difference compared with the Cistanche deserticola formula group (P ≤ 0.05).

[0027] Table 3 Results of the test on the contents of hepatic glycogen and muscle glycogen in the tissues of mice

[0028]

[0029] Note: a indicates significant difference compared with the blank control group (P ≤ 0.05), and b indicates significant difference compared with the Cistanche deserticola formula group (P ≤ 0.05).

[0030] The test results of Table 1, Table 2 and Table 3 show that, compared with the group using Cistanche deserticola formula alone (Group 1), the combination of protocorm-like bodies of Dendrobium huoshanense and Cistanche deserticola (Group 2) can significantly prolong the exhaustive swimming time of mice (P≤0.05), significantly reduce the content of urea nitrogen in the serum of mice (P≤0.05), significantly reduce the content of lactic acid in the serum of mice (P≤0.05), significantly increase the content of hepatic glycogen in the liver tissue (P≤0.05), and significantly increase the content of muscle glycogen in the muscle tissue (P≤0.05).

[0031] Example 3

[0032] Verification test on the effect of protocorm-like bodies of Dendrobium huoshanense in enhancing the kidney-tonifying and yang-strengthening effect of the formula of Cistanche deserticola, American ginseng, Rubus idaeus, Macrocarpium officinale and Lycium barbarum. Rats were used as experimental animals and randomly divided into a blank control group (Group 0), a kidney-yang deficiency model group (Group M), a Cistanche deserticola formula group (Group 1), and a combination of protocorm-like bodies of Dendrobium huoshanense and Cistanche deserticola group (Group 2). The blank control group and the kidney-yang deficiency model group were given distilled water by gavage, the Cistanche deserticola formula group was given the bionic extract of the Cistanche deserticola formula by gavage, and the combination of protocorm-like bodies of Dendrobium huoshanense and Cistanche deserticola group was given the bionic extract of the combination of protocorm-like bodies of Dendrobium huoshanense and Cistanche deserticola by gavage. The dosage of different extracts was 900 mg / kg, the gavage volume was 1.0 ml, and the drug was administered 2 times a day for 30 consecutive days.

[0033] Starting from the first day of the experiment, rats in the kidney-yang deficiency model group, the Cistanche deserticola formula group, and the combination of protocorm-like bodies of Dendrobium huoshanense and Cistanche deserticola group were subcutaneously injected with hydrocortisone injection, and the dosage of hydrocortisone was 10 mg / kg, once a day for 15 consecutive days, to prepare a kidney-yang deficiency rat model. On the 30th day after the first administration, the superoxide dismutase activity and malondialdehyde content in the serum were measured; the testosterone content in the serum was measured; the nitric oxide synthase activity in the penile tissue was measured; and the contents of nitric oxide and cyclic guanosine monophosphate in the penile tissue were measured.

[0034] The results of the verification test on the effect of protocorm-like bodies of Dendrobium huoshanense in enhancing the kidney-tonifying and yang-strengthening effect of the formula of Cistanche deserticola, American ginseng, Rubus idaeus, Macrocarpium officinale and Lycium barbarum are shown in Table 4, Table 5, Table 6 and Table 7.

[0035] Test results of superoxide dismutase activity and malondialdehyde content in the serum of rats in Table 4

[0036]

[0037] Note: c indicates significant difference from the model group (P≤0.05), and b indicates significant difference from the Cistanche deserticola formula group (P≤0.05).

[0038] Test results of testosterone content in the serum of rats in Table 5

[0039]

[0040] Note: c indicates significant difference from the model group (P≤0.05), and b indicates significant difference from the Cistanche deserticola formula group (P≤0.05).

[0041] Table 6 Test results of nitric oxide synthase activity in rat penile tissues

[0042]

[0043] Note: c indicates significant difference from the model group (P≤0.05), and b indicates significant difference from the Cistanche deserticola formula group (P≤0.05).

[0044] Table 7 Test results of nitric oxide content and cyclic guanosine monophosphate content in rat penile tissues

[0045]

[0046] Note: c indicates significant difference from the model group (P≤0.05), and b indicates significant difference from the Cistanche deserticola formula group (P≤0.05).

[0047] The test results in Table 4, Table 5, Table 6 and Table 7 show that, compared with the group using only Cistanche deserticola formula (Group 1), in the group of protocorm-like bodies of Dendrobium huoshanense + Cistanche deserticola formula (Group 2), the superoxide dismutase activity in rat serum increased significantly (P≤0.05), the malondialdehyde content in serum decreased significantly (P≤0.05), the testosterone content in serum increased significantly (P≤0.05), the nitric oxide synthase activity in penile tissues increased significantly (P≤0.05), the nitric oxide content in penile tissues increased significantly (P≤0.05), and the content of cyclic guanosine monophosphate in penile tissues increased significantly (P≤0.05).

[0048] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art in this technical field, without departing from the technical principle of the present invention, several improvements and modifications can still be made, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. The application of the protocorm of Dendrobium huoshanense in anti-fatigue and kidney-tonifying and yang-strengthening is characterized in that The protocorm of Dendrobium huoshanense can significantly improve the anti-fatigue, kidney-tonifying and yang-enhancing effects of the formula composed of Cistanche deserticola, American ginseng, raspberry, Cornus officinalis and Lycium barbarum.

2. The use of the protocorm of Dendrobium huoshanense in anti-fatigue and kidney-tonifying and yang-strengthening according to claim 1, characterized in that The Huoshan Dendrobium protocorm is an undifferentiated Huoshan Dendrobium protocorm, and the differentiated Huoshan Dendrobium buds or Huoshan Dendrobium plants are not found to have the effect of improving the anti-fatigue, kidney-tonifying and yang-strengthening effects of the formula.

3. The use of the protocorm of Dendrobium huoshanense in anti-fatigue and kidney-tonifying and yang-strengthening according to claim 1, characterized in that The application method comprises the following steps: (1) Add 3 parts by weight of dry protocorm of Dendrobium huoshanense to a formula of 5 parts by weight of Cistanche deserticola, 5 parts by weight of American ginseng, 3 parts by weight of raspberry, 2 parts by weight of Cornus officinalis, and 1 part by weight of wolfberry, and grind to powder for later use; (2) The ultrafine powder was added to a gastric juice simulation solution with a pH of 2.0 and a pepsin activity of 2000 U / mL at a solid-liquid ratio of 1:50, and stirred at 37°C for 2 h. The pH was adjusted to 6.8 with a 1 mol / L NaOH solution to inactivate pepsin. Then, trypsin and pancreatic amylase were added until the activities were both 100 U / mL. Stirring was continued at 37°C for 2 h, and then heated to 95°C for 5 min to inactivate the enzymes. The supernatant was filtered and concentrated and dried to obtain a bionic extract. (3) Bionic extracts can be used in traditional Chinese medicine preparations or functional foods for anti-fatigue and kidney-tonifying and aphrodisiac effects.

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