TWI869529B - The pharmaceutical composition of dendrobium for ameliorating muscle wasting caused by cachexia and manufacture thereof - Google Patents

Abstract

The present invention provides a composition for use in the amelioration of muscle wasting and muscle deficiency. The composition comprising an effective amount of Dendrobium Taiseed Tosnobile (Taiseed Emperor NO.1) extract and pharmaceutically acceptable carrier, excipients or salts. The muscle wasting and muscle deficiency include those caused by tumor cachexia or aging.

Description

A pharmaceutical composition of dendrobium for alleviating muscle wasting caused by cachexia and its preparation

The present invention provides a composition for alleviating muscle wasting caused by cachexia, the composition comprising an effective dose of a dendrobium extract and a pharmaceutically acceptable carrier, excipient or salt. Muscle wasting is mainly caused by disease or aging.

The body's muscle mass reaches its peak at around 25 years old, and then begins to decrease slowly. Between the ages of 40 and 70, the weight decreases by about 40%. The most obvious turning point occurs after the age of 60, when the rate of skeletal muscle loss increases, which also speeds up the rate of weakness.

Especially in the process of aging, the causes of muscle atrophy are complex and diverse, including the re-innervation of motor neurons with aging, the decrease in protein synthesis rate and the decrease in related hormones, which may all be the causes of muscle atrophy. In addition, muscle loss is often accompanied by an increase in body fat and a decrease in metabolic rate, which leads to metabolic syndromes such as obesity and type 2 diabetes. The decrease in muscle strength and mobility increases the risk of falls and fractures, making the elderly lose the ability to live independently. It is estimated that the cost of muscle atrophy in the United States is about 11.8 billion to 26.2 billion US dollars each year. If the occurrence of sarcopenia can be properly prevented and treated, considerable expenses can be saved each year. With the advancement of medicine, the average life expectancy of modern people is getting longer and longer. At present, the elderly population (over 65 years old) in our country has exceeded 14% of the total population in 2017, and will further become an aging society. For us who are gradually moving towards an aging society, how to use drugs to prevent and reduce the occurrence of muscle atrophy and loss in the elderly, so as to improve the quality of life of the elderly and reduce the cost of medical treatment and care is indeed a very important issue at present.

In addition, according to NCI (National Cancer Institute According to the National Cancer Institute, nearly 50% of cancer patients will have symptoms of cachexia, which makes their nutritional status very poor. Cachexia is the result of abnormal hormones and metabolism in cancer patients, or reduced food intake. Cancer patients have various symptoms of loss of appetite or weight loss. Usually when patients begin to lose weight, their muscles appear to be atrophied, their activity level decreases, and they become tired and weak, cancer cachexia will significantly reduce the survival rate and quality of life of cancer patients. It is characterized by a decrease in muscle mass and weight. In addition to loss of appetite, reduced muscle protein synthesis, increased muscle protein degradation, and increased cell apoptosis will all lead to a decrease in muscle mass in cachectic patients. However, the mechanism of muscle atrophy caused by cancer cachexia is still not very clear.

The mechanism of muscle loss caused by the disease is not very clear. At present, the common treatment for muscle atrophy is physical therapy, but physical therapy adopts methods such as increasing muscle strength training and walking training. However, cancer patients or elderly patients with muscle atrophy are not suitable for forced physical exercise, and there are many doubts about the effectiveness of treatment. There is no effective drug or food to treat or improve symptoms. Therefore, there is still a great need for a safe drug or food with low side effects that can be used by cancer patients or elderly patients.

To solve the above problems, the present invention uses a new preparation method to make dendrobium for relieving muscle wasting. Dendrobium nobile (scientific name: Dendrobium nobile ) is a species of the genus Dendrobium of the Orchidaceae family. It is also known as Xianhu Lanyun, Immortal Grass, Resurrection Grass, Purple Fairy Plant, Chlorophytum, Forest Orchid, Forbidden Life, Golden Hairpin Flower, Golden Hairpin Dendrobium, Golden Hairpin, Yellow Grass, Golden Silk Dendrobium. The stem is upright, fleshy and thick, slightly flat and cylindrical, 10 to 60 cm long and 1.3 cm thick. It is a medicinal plant with a sweet and slightly salty taste, cold, and enters the stomach, kidney, and lung meridians. It benefits the stomach and produces fluid, nourishes yin and clears heat. It is used for yin damage and fluid deficiency, dry mouth and thirst, poor appetite and dry vomiting, deficiency heat after illness, and blurred vision. Dendrobium is widely used to treat kidney, lung and gastrointestinal diseases. Currently, Dendrobium has not been used alone to treat muscle loss.

The main purpose of the present invention is to provide a pharmaceutical composition using a Dendrobium extract for preparing a pharmaceutical composition for relieving muscle wasting caused by cachexia.

In one embodiment of the present invention, the dendrobium extract is obtained from a plant of the genus Dendrobium.

In one embodiment of the present invention, the Dendrobium plant is Dendrobium officinale.

In one embodiment of the present invention, the Dendrobium extract comprises a water extract of Dendrobium chrysantha or a wine extract of Dendrobium chrysantha.

In one of the best embodiments of the present invention, the Dendrobium extract is a water extract of Dendrobium officinale.

In one embodiment of the present invention, the Dendrobium extract comprises a water extract, a 50% alcohol extract or a 95% alcohol extract.

In one embodiment of the present invention, the scope of use includes improving muscle atrophy, protecting muscle cell differentiation, increasing muscle grip strength, increasing survival rate or increasing muscle mass.

In one embodiment of the present invention, the muscle wasting is caused by tumor malignancy.

In one embodiment of the present invention, the effective dosage concentration of the Dendrobium officinale water extract is 100mg/ml~400mg/ml.

In one of the best embodiments of the present invention, the effective dosage concentration of the Dendrobium officinale water extract is 200 mg/ml.

In one embodiment of the present invention, the effective dosage concentration of the Dendrobium water extract is 50μg/ml~200μg/ml.

In one of the best embodiments of the present invention, the effective dosage concentration of the Dendrobium water extract is 50μg/ml.

In one embodiment of the present invention, the effective dosage of the water extract of Dendrobium officinale is 75-150 mg/kg, and the effective dosage of the wine extract of Dendrobium officinale is 100-200 mg/kg.

In one embodiment of the present invention, the pharmaceutical composition comprises a dendrobium extract and a pharmaceutically acceptable carrier, excipient or salt.

The present invention provides a method for using a dendrobium extract in the preparation of a pharmaceutical composition for treating and relieving muscle wasting caused by cachexia, wherein the dendrobium extract is prepared by a method comprising the following steps:

(1) The dried and powdered Dendrobium sample was thoroughly mixed with pure water, and then the resulting mixture was shaken with an ultrasonic oscillator for about 60-90 minutes, and then filtered to collect the filtrate. The residue obtained by filtration was used to repeat the above steps of pure water mixing-ultrasonic vibration-filtration three times in total. Then all the collected filtrate was decompressed and concentrated to remove the pure water, thereby obtaining the Dendrobium water extract.

(2) The residue of the water extraction of Dendrobium officinale in step (1) is subjected to ultrasonic vibration extraction, filtration and concentration according to the extraction steps of step (1) above, to obtain a 50% alcohol extract of Dendrobium officinale.

(3) The residue of the dendrobium wine extraction in step (2) is subjected to ultrasonic vibration extraction, filtration and concentration according to the extraction steps in step (1) above with 70% alcohol to obtain a 70% alcohol extract of dendrobium.

(4) The residue of the dendrobium wine extraction in step (3) is subjected to ultrasonic vibration extraction, filtration and concentration according to the extraction steps in step (1) above, to obtain a 95% alcohol extract of dendrobium.

In one embodiment of the present invention, the method described, wherein the dendrobium extract is taken from Dendrobium officinale.

In the method described in one embodiment of the present invention, the Dendrobium extract comprises a Dendrobium water extract or a Dendrobium wine extract.

In the method described in one of the best embodiments of the present invention, the Dendrobium extract is a Dendrobium water extract.

In one embodiment of the present invention, the method described, wherein the Dendrobium extract comprises a water extract, a 50% alcohol extract or a 95% alcohol extract.

The attached diagrams illustrate one or more examples of this case, and together with the relevant explanatory content, they are used to explain the principle of this case.

Figure 1 shows the experimental flow chart showing that both water extract and wine extract of Dendrobium can restore muscle loss caused by tumor malignancy.

Figures 2 to 6 show that both water extracts and wine extracts of Dendrobium can restore muscle wasting caused by tumor cachexia, restore muscle function and increase the survival rate of tumors, and the high-dose water extract group is the group with the best effect. Figure 2 shows the effect of different Golden Emperor Dendrobium extracts on regulating the body weight of tumor mice; Figure 3 shows the effect of different Golden Emperor Dendrobium extracts on increasing muscle grip strength of tumor mice; Figure 4 shows the effect of different Golden Emperor Dendrobium extracts on antagonizing muscle wasting in tumor mice; Figure 5 shows the effect of tissue staining of thigh cross-sections; Figure 6 shows the regulatory effect of different Golden Emperor Dendrobium extracts on cachexia-related cytokines.

Figure 7 shows the experimental groups and flow chart of Dendrobium water extract and wine extract protecting muscle cells (wherein the concentration of Dendrobium water extract is 0, 200 or 400μg/ml; the concentration of Dendrobium wine extract is 0, 100 or 200μg/ml); the experimental groups and flow chart of potential target genes related to Dendrobium water extract (concentration is 200μg/ml) protecting muscle cells.

Figure 8 shows that at the beginning of differentiation, bleomycin and different concentrations of Dendrobium wine extract (concentrations of 0, 200 or 400 μg/ml) were added to differentiated muscle cells at the same time. Four days later, the muscle cell fusion rate was analyzed by giemsa staining, indicating that Dendrobium wine extract protected muscle cells.

Figure 9 shows that bleomycin and different concentrations of Dendrobium water extract (concentrations of 0, 200 or 400 μg/ml) were added to differentiated muscle cells at the beginning of differentiation. Four days later, the muscle cell fusion rate was analyzed by giemsa staining, indicating that Dendrobium water extract directly protected muscle cells.

Figure 10 shows the results of inducing differentiation of myofibroblast cell lines (C2C12 cells) with low concentration horse serum (2% HS) and simultaneously treating them with bleomycin (BLM) and Dendrobium water extract (concentration of 200μg/ml). RNA of the control group and experimental group cell lines was collected on the eighth day for next generation sequencing reaction and gene expression profiling analysis.

Figure 12 shows the effect of Dendrobium water extract on the growth of myofibroblasts. As shown in the figure, Dendrobium water extract can reverse the death of myofibroblasts caused by bleomycin treatment (the survival rate increased from 60% to 90%), which means that it has the effect of protecting muscle cells.

Figure 13 shows the effects of 50% alcohol extract and 95% alcohol extract of Dendrobium officinale wine extract on the growth of myofibroblasts. In addition, this result also shows that the effective ingredients in Dendrobium officinale water extract have a direct effect and protect myofibroblasts.

Figure 14 shows the three extract groups (including water extract, 50% alcohol extract and 95% alcohol extract) of Dendrobium officinale extract in the protection experiment of myofibroblast differentiation into myotubes (each group contains concentrations of 0, 25, 50, or 100μg/ml) and the flow chart.

Figures 15-17 show that we analyzed the effects of different extracts from Dendrobium officinale extract on protecting myofibroblasts from differentiation into myotubes after bleomycin treatment.

Figure 18 shows the experiment of Dendrobium water extract and alcohol extract protecting mice from muscle atrophy caused by tumors, in which three extract groups of Dendrobium extract (including water extract, 50% alcohol extract or 95% alcohol extract) were divided into groups (water extract concentration was 25 or 50 mg/kg, 50% alcohol extract concentration was 25 or 50 mg/kg, 95% alcohol extract concentration was 10 or 25 mg/kg) and the flow chart.

Figures 19-21 show the tumor malignancy model of mice injected subcutaneously with LLC cell lines in animal experiments. One week after the LLC cell lines were injected, different concentrations of Dendrobium extracts were fed for 4 weeks, 5 days a week. After 4 weeks, the malignancy phenomena induced by the tumor, such as muscle atrophy, were observed to be improved.

Example 1: Preparation of Dendrobium officinale Extract

(1) The dried and powdered golden huang shi huang sample was thoroughly mixed with pure water, and then the resulting mixture was shaken with an ultrasonic oscillator for about 60-90 minutes, and then filtered to collect the filtrate. The residue obtained by filtration was used to repeat the above pure water mixing-ultrasonic vibration-filtration steps three times in total. Then all the collected filtrate was decompressed and concentrated to remove the pure water, thereby obtaining the golden huang shi huang water extract.

(2) The residue of the golden huang shi huang extract in step (1) is subjected to ultrasonic vibration extraction-filtration-concentration according to the extraction steps of step (1) above, and a 50% alcohol extract of golden huang shi huang can be obtained.

(3) The residue of the golden huang shi huang extract in step (2) is subjected to ultrasonic vibration extraction, filtration and concentration according to the extraction steps of step (1) above with 70% alcohol to obtain a golden huang shi huang 70% alcohol extract.

(4) The residue of the golden huang shi huang extract in step (3) is subjected to ultrasonic vibration extraction, filtration and concentration according to the extraction steps of step (1) above, and a golden huang shi huang 95% alcohol extract can be obtained.

Example 2: Experiment on mice with weight loss and muscle wasting caused by tumor malignancy

After one week of acclimatization, all C57BL/6 mice except the normal group were given Lewis lung cancer cells by subcutaneous injection on the first day to induce tumor formation. They were then randomly divided into the following 7 groups (see Table 1 below). The mice were orally fed with the extract of Dendrobium chrysantha of each group for 5 days a week for 4 weeks. Finally, the mice were sacrificed, and the relevant weights and tissue staining were calculated. Figures 2-6 show that the water extract or wine extract of Dendrobium chrysantha can restore the weight loss and muscle loss caused by tumor malignancy.

As shown in Figures 2 to 6, the water extract or wine extract of Dendrobium officinale can restore weight loss and muscle wasting caused by tumor malignancy, and can restore muscle function. The low-dose groups of water extract or 95% wine extract (giving 75 or 150 mg/kg B.W. of Dendrobium officinale water extract; 100 mg/kg B.W. of 95% wine extract) are both very effective groups. The results in Figure 2 show the effects of different groups of Dendrobium chrysantha extracts in Table 1 on the average weight of tumor mice, among which the water extract and 95% wine extract (giving 75 or 150 mg/kg B.W. of Dendrobium chrysantha water extract; 100 or 200 mg/kg B.W. of 95% wine extract) can effectively increase the weight of tumor mice; and the results in Figure 3 show the effects of different Dendrobium chrysantha extracts on increasing the muscle grip strength of tumor mice, among which the low-dose groups of water extract or 95% wine extract (giving 75 or 150 mg/kg B.W. of Dendrobium chrysantha water extract; 100 mg/kg B.W. of 95% wine extract) are the groups with the best effects and are closest to the control group values; the leg appearance results in Figure 4 show the effects of different Dendrobium chrysantha extracts in antagonizing muscle wasting in tumor mice. Figure 5 is a cross-section of the thigh tissue stained by mice sacrificed 27 days after tumor cell injection, showing the protective effect of different Dendrobium chrysanthemi extracts on muscle atrophy caused by tumor. Figure 6 is a blood sample taken from mice before they were sacrificed 27 days after tumor cell injection and analyzed for related cytokines by ELISA, showing the regulatory effect of different Dendrobium chrysanthemi extracts on the expression of cytokines related to cachexia or muscle regeneration.

Example 3: Experiment on the protection of C2C12 cells by Dendrobium officinale extract

C2C12 cells were seeded into 24-well culture dishes on the first day. Low-concentration horse serum (2%) was added on the second day to induce differentiation into muscle cells and produce fiber bundles. On the fourth day, low-concentration horse serum (2%) was replaced and bleomycin (a cancer drug) and different concentrations of water extract of Dendrobium chrysanthemi (concentrations: 0, 200, 400 μg/ml) or 95% alcohol extract (concentrations: 0, 100, 200 μg/ml) were added to the differentiated muscle cells. Every six days, low-concentration horse serum (2%) was replaced and bleomycin and different concentrations of water extracts of Dendrobium chrysanthemi (concentrations: 0, 200, 400μg/ml) or 95% alcohol extracts (concentrations: 0, 100, 200μg/ml) were added to the differentiated muscle cells. On the eighth day, the cells were collected and the muscle cell fusion rate was evaluated by giemsa staining to establish the attenuation pattern of muscle cell fiber bundles. The experimental groups and process and the dosage diagram of Dendrobium chrysanthemi types are shown in Figure 7.

The results of the protection of muscle cells by Dendrobium chinense extract are shown in Figure 8. At the beginning of differentiation, bleomycin (a cancer treatment drug) and different concentrations of Dendrobium chinense 95% wine extract (concentrations: 0, 100, 200 μg/ml) were added to the differentiated muscle cells. Four days later, the muscle cell fusion rate was analyzed by giemsa staining, showing that the muscle cell fusion rate of Dendrobium chinense 95% wine extract (concentrations: 0, 100, 200 μg/ml) increased. Add, but there is no concentration dependence; as shown in Figure 9, at the beginning of differentiation, bleomycin (cancer treatment drug) and different concentrations of water extract of Dendrobium chrysanthemi (concentration: 0, 200, 400μg/ml) were added to the differentiated muscle cells at the same time. Four days later, the muscle cell fusion rate was analyzed by giemsa staining, showing that the muscle cell fusion rate of water extract of Dendrobium chrysanthemi (concentration: 100, 200μg/ml) increased, but there was no concentration dependence.

The potential target gene process related to the protection of muscle cells by Dendrobium officinale extract is shown in Figure 7. The myofibroblast cell line (C2C12 cells) was incubated with low concentration of horse serum (2%) HS) was used to induce differentiation and simultaneously treated with bleomycin (a cancer treatment drug) and water extract of Dendrobium chrysanthemi. On the eighth day, RNA from the control group and experimental group cell lines was collected for next generation sequencing. The experimental groups were divided into: (1) undifferentiated control group, (2) differentiated group, (3) differentiated group plus bleomycin (BLM: 100 μg/ml), and (4) differentiated group plus bleomycin plus water extract of Dendrobium chrysanthemi (BLM: 100 μg/ml; DTT-W: 200 μg/ml). Next, the gene expression profiles of these four groups were separated and analyzed separately. A total of 22,195 gene expression data were obtained. In these data, we searched for the target gene expression The current amount is consistent with the trend of (1) undifferentiated control group and (3) differentiation plus bleomycin group (BLM: 100μg/ml), while it is opposite to (2) differentiation group and (4) differentiation plus bleomycin group plus Dendrobium officinale water extract group (BLM: 100μg/ml; DTT-W: 200μg/ml). This means that this target gene will be highly related to myofibroblast differentiation and will also change with the treatment of bleomycin or Dendrobium officinale water extract. After this conditional screening, the results are shown in Figure 10. We obtained a total of 213 potential target gene groups. We then further analyzed the muscle-related gene groups in these gene groups. After literature comparison, we finally found 7 target gene groups that are potentially affected by bleomycin and water extract of Dendrobium officinale.

Example 4: Cell experiment on the effect of Dendrobium officinale extract on protecting muscle cells

On the first day, myofibroblast cell lines (C2C12 cells) were seeded into 24-well culture dishes. On the second day, extracts of Dendrobium chrysanthemi (water extract, 50% alcohol extract, and 95% alcohol extract) and bleomycin were added. On the fourth day, cells were collected and cell survival rate was calculated by MTT assay, as shown in the flow chart in Figure 11.

The effects of Dendrobium chrysanthemi extracts (water extract, 50% alcohol extract and 95% alcohol extract) on the growth of myofibroblasts are shown in Figures 12 and 13. The results in Figure 12 show that the water extract of Dendrobium chrysanthemi can reverse the death of myofibroblasts caused by bleomycin treatment (cell survival rate increased from 60% to 90%), and the water extract of Dendrobium chrysanthemi has the effect of protecting muscle cells; and the results in Figure 13 show that the 50% alcohol extract and the 95% alcohol extract of Dendrobium chrysanthemi wine extract have the effect of protecting muscle cells. In contrast, the treatment with alcohol extract could not reverse the toxic effect of bleomycin on myofibroblasts, which means it had no protective effect. What's more, the treatment with 95% alcohol extract of Golden Emperor Dendrobium wine extract further enhanced the toxic effect of bleomycin on myofibroblasts (the cell survival rate dropped from 40% to 18%). Therefore, the 95% alcohol extract of Golden Emperor Dendrobium wine extract may contain ingredients that inhibit the growth of myofibroblasts. In addition, these results also show that the water extract of Golden Emperor Dendrobium has effective ingredients that directly act and protect myofibroblasts.

Example 5: Cell experiment on the effects of three extracts of Dendrobium officinale extract on differentiation of fibroblasts into myotubes

C2C12 cells were seeded into 24-well culture dishes on the first day. Low concentration horse serum (2%) was added on the second day to induce differentiation into muscle cells and produce fiber bundles. On the fourth day, low concentration horse serum (2%) was replaced with bleomycin (a cancer drug) and different concentrations of Dendrobium chrysanthemi extracts (water extract, 50% alcohol extract, and 95% alcohol extract) were added at the same time. Extract) groups (each group contained 0, 25, 50 or 100 μg/ml) to differentiated muscle cells. On the sixth day, low concentration horse serum (2%) was replaced and bleomycin and different concentrations of Dendrobium chrysanthemi extract were added to differentiated muscle cells at the same time. On the eighth day, cells were collected to detect the differentiation ratio of muscle cells, as shown in the flow chart in Figure 14.

The effects of three groups of Dendrobium officinale extracts (water extract, 50% alcohol extract and 95% alcohol extract) (each group contained 0, 25, 50 or 100 μg/ml) on protecting myofibroblasts from differentiating into myotubes after bleomycin treatment are shown in Figures 15-17. The results of water extract in FIG. 15 , 50% alcohol extract in FIG. 16 , and 95% alcohol extract in FIG. 17 respectively show the protective effect on the differentiation of myofibroblasts into myotubes. The results of FIG. 15 show that when myofibroblasts are pretreated with a specific concentration of Dendrobium chrysanthemi water extract, the muscle cell differentiation inhibition caused by bleomycin can be effectively alleviated. This result shows that the extract contains effective ingredients that can directly act on myofibroblasts to counteract the differentiation inhibition of bleomycin. Interestingly, the results in Figure 16 show that the treatment with 50% alcohol extract cannot protect the inhibitory effect of bleomycin on myofibroblast differentiation. What's more, the results in Figure 17 show that the treatment with 95% alcohol extract can promote the ability of bleomycin to inhibit the differentiation of myofibroblasts into myotubes, indicating that there may be components in the 95% alcohol extract that inhibit the growth and differentiation of myofibroblasts.

Example 6: Experiment on the effect of three extracts of Dendrobium officinale on protecting mice from muscle atrophy caused by tumor

After C57BL/6 mice were raised for one week to adapt to the environment, mice in the tumor groups except the healthy group were given Lewis lung cancer cells by subcutaneous injection on the first day to induce tumor formation. Then they were randomly divided into the following 8 groups and orally fed with the extract of Dendrobium chrysantha of each group 5 days a week for 4 weeks. Finally, the mice were sacrificed, the relevant weights were calculated, and the tissues were stained. As shown in Figure 18, in the experiment of Dendrobium chrysantha water extract and wine extract protecting mice from muscle atrophy caused by tumor, the groups were divided into the following groups:

1. Healthy group

2. Tumor group

3. Tumor plus low-dose group of water extract of Dendrobium officinale (25mg/kg, abbreviated as: Dwfw-L)

4. Tumor plus high-dose group of water extract of Dendrobium officinale (50mg/kg, abbreviated as: Dwfw-H)

5. Tumor plus low-dose group of 50% alcohol extract of Dendrobium officinale (25mg/kg, abbreviated as: Dwfe50-L)

6. Tumor plus high-dose group of 50% alcohol extract of Dendrobium officinale (50mg/kg, abbreviated as: Dwfe50-H)

7. Tumor plus low-dose group of 95% alcohol extract of Dendrobium officinale (10mg/kg, abbreviated as: Dwfe95-L)

8. Tumor plus high-dose group of 95% alcohol extract of Dendrobium officinale (20mg/kg, abbreviated as: Dwfe95-H)

The results of the three extracts of Dendrobium chrysantha on protecting against muscle atrophy caused by tumors are shown in Figures 19-21. In animal experiments, LLC cell lines were first injected subcutaneously into mice to induce tumor malignancy. One week after the LLC cell lines were injected, the mice were fed with Dendrobium chrysantha extracts of different concentrations for 4 weeks. Every week for 5 days, it was observed whether the malignancy phenomena induced by tumors, such as muscle atrophy, were improved. As shown in Figures 19-21, the results in Figure 19 show that the gastrocnemius muscle of the tumor group The weight of the gastrocnemius muscle (also called explosive muscle) was significantly reduced. The results in Figure 20 show that the grip strength test of the tumor group also showed a significant decrease, which is one of the characteristics of cachexia. On the other hand, in the results in Figure 19, after feeding tumor mice with water extract of Dendrobium officinale, the weight of the gastrocnemius muscle showed an increasing trend, and the results in Figure 20 showed that its grip strength also increased. This phenomenon does not seem to have a dose-dependent effect. On the other hand, 50% and 95% alcohol extracts Feeding of different extracts can also have the effect of restoring muscle mass and function, and it is also not dose-dependent. In addition, the results of hind limb tissue staining in Figure 21 show that the rectus and peroneus longus muscles (indicated by arrows) of the hind limbs of tumor mice are indeed significantly reduced compared with the control group, and after feeding different extracts, the rectus and peroneus longus muscles are increased. These results show that different extracts of Dendrobium chrysanthemi have effective ingredients that can reduce the degree of muscle atrophy in cachexia. Interestingly, we found that mice injected with tumor cells had obvious cardiac hypertrophy on the 21st day, and the tissue sections showed that the ventricles were enlarged and the heart muscle was reduced. Similarly, feeding with water extract of Dendrobium officinale, 50% and 95% alcohol extracts can also effectively alleviate this pathological phenomenon.

Although the present invention has been described in specific exemplary embodiments and examples, it should be understood that the embodiments disclosed herein are for illustrative purposes only and can be modified and altered by those skilled in the art without departing from the spirit and scope of the present invention as described in the following claims.

The most important thing is that the research on animals and cells found that the Dendrobium officinale extract can effectively improve muscle atrophy, protect muscle cell differentiation, increase muscle grip strength, increase survival rate and increase muscle mass. In summary, the Dendrobium officinale extract produced by the present invention is an excellent drug ingredient to improve muscle loss caused by diseases.

Claims (6)

A use of a dendrobium extract in preparing a pharmaceutical composition for treating and relieving muscle wasting caused by cachexia, wherein the pharmaceutical composition is composed of a dendrobium extract and an active ingredient, the dendrobium extract is a Dendrobium Taiseed Tosnobile extract, wherein the Dendrobium Taiseed Tosnobile extract comprises a Dendrobium Taiseed Tosnobile water extract, a Dendrobium Taiseed Tosnobile 50% alcohol extract or a 95% alcohol extract; the active ingredient is selected from the group consisting of total polysaccharides, total alkaloids, total polyphenols, total flavonoids, total saponins and total flavonoids, wherein the cachexia is a tumor mediated by the IL-6 signaling pathway, and wherein the pharmaceutical composition is administered to a subject in need at an effective dose. The use as described in item 1 of the patent application scope, wherein the scope of the use includes improving muscle atrophy, protecting muscle cell differentiation, increasing muscle grip strength, increasing survival rate or increasing muscle mass. The use as described in item 1 of the patent application, wherein the administration method includes oral administration. For the use described in item 1 of the patent application, the effective dosage of the water extract of Dendrobium officinale is 75-150 mg/kg, and the effective dosage of the wine extract of Dendrobium officinale is 100-200 mg/kg. The use as described in item 1 of the patent application, wherein the pharmaceutical composition comprises the Dendrobium extract and a pharmaceutically acceptable carrier, excipient or salt. The use as described in item 1 of the patent application, wherein the preparation method of the dendrobium extract comprises the following steps: (1) thoroughly mixing a dried and powdered dendrobium sample with pure water, then vibrating the resulting mixture with an ultrasonic vibrator for about 60-90 minutes, then filtering, and then collecting the filtrate. The residue obtained by filtration is used to repeat the above-mentioned pure water mixing-ultrasonic vibration-filtration steps three times in total, and then all the collected filtrate is decompressed and concentrated to remove the pure water, thereby obtaining the dendrobium water extract; ( 2) The residue of the Dendrobium extract in step (1) is subjected to ultrasonic vibration extraction, filtration and concentration according to the extraction steps in step (1) with 50% alcohol to obtain a 50% alcohol extract of Dendrobium; (3) The residue of the Dendrobium extract in step (2) is subjected to ultrasonic vibration extraction, filtration and concentration according to the extraction steps in step (1) to obtain a 70% alcohol extract of Dendrobium; (4) The residue of the Dendrobium extract in step (3) is subjected to ultrasonic vibration extraction, filtration and concentration according to the extraction steps in step (1) to obtain a 95% alcohol extract of Dendrobium.