Use of Desmodium renifolium and its extract in the preparation of a medicament for treating sarcopenia

Abstract

The present invention discloses the use of Desmodium renifolium and its extract in the preparation of a medicament for treating sarcopenia. Based on the muscle injury model experiment of Caenorhabditis elegans, the present invention determines that the extract of Desmodium renifolium can reverse the decline in motor function caused by muscle injury, and at the same time, the body wall muscle can also be restored to a certain extent, and the overall lifespan of the nematode can be extended. Applying Desmodium renifolium to the treatment of sarcopenia has broad application prospects.

Description

Technical Field

[0001] The present invention specifically relates to the use of Desmodium renifolium (Linn.) Schindl and its extracts in the preparation of drugs for treating sarcopenia. Background Art

[0002] As the body's protein repository and the largest organ system, skeletal muscle is the material basis for providing support and balance functions and performing physical activities. When neuromuscular degenerative changes mainly manifested by muscle mass reduction, muscle strength and function decline occur in skeletal muscle due to various reasons, it will trigger adverse events such as limited daily activities, falls, and disabilities of the body, reduce the quality of life, and increase the risk of death. This change mainly occurs after entering the aging process with age, affected by factors such as changes in hormone levels, imbalance between protein synthesis and catabolism, mitochondrial chromosome damage, free radical oxidative damage, impaired repair mechanism of skeletal muscle, apoptosis, and calcium homeostasis imbalance, and is related to the body's lifestyle, exercise habits, diseases, and nutritional status. This phenomenon is ultimately described as "sarcopenia". With the continuous aggravation of the aging degree of the population in China and the world, the number of elderly people is increasing day by day, the incidence of sarcopenia is increasing, and a series of health problems such as falls, disabilities, loss of labor force and activity ability of the elderly have attracted extensive attention.

[0003] Desmodium renifolium (Linn.) Schindl is a plant of the genus Desmodium in the family Leguminosae of the order Rosales, and it is one of the commonly used plant medicines of the Dai people in Xishuangbanna, Yunnan. It has the effects of clearing away heat and detoxifying, relieving cough and resolving phlegm, promoting qi circulation and relieving pain, diuretic and jaundice-reducing, and tonifying qi and enhancing sexual function. It is used for headache in hot-season colds, sore throat, cough and asthma, jaundice in bile diseases, and being weak and sickly. At present, there is no report on the treatment of sarcopenia with Desmodium renifolium, and there is no application in improving muscle function decline. Summary of the Invention

[0004] To solve the above problems, the present invention provides the use of Desmodium renifolium and its extracts in the preparation of drugs for treating sarcopenia.

[0005] Furthermore, the extract is an ethanol extract of Desmodium renifolium.

[0006] Furthermore, the extract is a 95% ethanol extract of Desmodium renifolium.

[0007] Furthermore, the 95% ethanol extract is prepared by refluxing Desmodium renifolium with 95% ethanol and removing the ethanol from the extract.

[0008] Further, the drug is a drug for improving the decline of motor function.

[0009] Furthermore, the drug is a drug for improving the decline in motor function caused by muscle injury.

[0010] Further, the drug is a drug for repairing damaged muscles.

[0011] Further, the drug is a drug for improving the shortening of lifespan caused by the decline in motor ability.

[0012] Further, the drug is a preparation prepared by taking Desmodium renalatum or its extract as an active ingredient and adding pharmaceutically acceptable excipients.

[0013] Furthermore, the preparation is an oral preparation, an external preparation or an injectable preparation.

[0014] Furthermore, the oral preparation is a granule, a solution, a pill, a capsule or a powder; the external preparation is a plaster, a spray, a liniment or an emulsion; the injectable preparation is an injectable solution or a powder for injection.

[0015] The use of Desmodium renalatum and its extract of the present invention in the preparation of a drug for treating sarcopenia. Based on the muscle injury model experiment of Caenorhabditis elegans, it is determined that the extract of Desmodium renalatum can reverse the decline in motor function caused by muscle injury, and at the same time, the body wall muscles can also be restored to a certain extent, and the overall lifespan of the nematodes can be extended. Applying the extract of Desmodium renalatum to the treatment of sarcopenia, especially the muscle injury and the decline in motor function caused by muscle injury during the course of sarcopenia, has broad application prospects.

[0016] Obviously, according to the above content of the present invention, according to the common general knowledge and conventional means in the art, without departing from the above basic technical idea of the present invention, various other forms of modification, substitution or change can also be made.

[0017] The above content of the present invention will be further described in detail below through specific embodiments in the form of examples. However, this should not be construed as limiting the scope of the above subject matter of the present invention to the following examples. All technologies implemented based on the above content of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Effects of Desmodium renalatum extracts at different concentrations on the swimming frequency of dexamethasone-induced Caenorhabditis elegans (n = 30);

[0019] Figure 2 Effects of Desmodium renalatum extracts at different concentrations on the crawling frequency of dexamethasone-induced Caenorhabditis elegans (n = 30)

[0020] Figure 3Effects of Desmodium renifolium extracts at different concentrations on the head swing of Caenorhabditis elegans damaged by dexamethasone (n = 30)

[0021] Figure 4 Effects of Desmodium renifolium extracts at different concentrations on the pharyngeal pumping of Caenorhabditis elegans damaged by dexamethasone (n = 30)

[0022] Figure 5 Effects of Desmodium renifolium extracts at different concentrations on the lifespan of Caenorhabditis elegans damaged by dexamethasone (n = 3)

[0023] Figure 6 Effects of Desmodium renifolium extracts at different concentrations on the reproductive ability of Caenorhabditis elegans damaged by dexamethasone (n = 8)

[0024] Figure 7 Rhodamine-phalloidin staining images of the body wall muscles of Caenorhabditis elegans in each group (n = 10) Detailed implementation manners

[0025] The raw materials, equipment and reagents used in the detailed implementation manners of the present invention are all known products and are obtained by purchasing commercially available products.

[0026] Example 1 Application research on treating sarcopenia with Dai medicine Desmodium renifolium

[0027] 1. Materials

[0028] 1.1 Experimental reagents and instruments

[0029] Desmodium renifolium (Linn.) Schindl (provided by the Key Laboratory of Dai and Yi Medicine of Yunnan Province and identified as authentic by Senior Experimentalist Yin Zili of Yunnan University of Traditional Chinese Medicine); Dexamethasone Sodium Phosphate (2204092211); Peptone (P000000400), Yeast Extract (P000000406), Beijing Baierdi Biotechnology Co., Ltd.; Agar Powder (112M021), Beijing Solarbio Science & Technology Co., Ltd.; Cholesterol (C804517), 5-Fluoro-2'-deoxyuridine (FUDR) (F809658), Disodium Hydrogen Phosphate Dodecahydrate (C12067867), Shanghai Macklin Biochemical Co., Ltd.; Anhydrous Calcium Chloride (20201204), Anhydrous Magnesium Sulfate (20200804), Sodium Chloride (20210406), Potassium Dihydrogen Phosphate (20210104), Dipotassium Hydrogen Phosphate (20201204), Sodium Hydroxide (20210104), Tianjin Fengchuan Chemical Reagent Technology Co., Ltd.; Sodium Hypochlorite (20220801), Tianjin Damao Chemical Reagent Factory; DMSO (20221201514), Zhiyuan Reagent Chemical Co., Ltd. (Tianjin); Triton X-100 (N0414A), Meilun Biotechnology Co., Ltd.; Methyl Viologen (Paraquat) (F2123262), Shanghai Aladdin Biochemical Technology Co., Ltd.; PBS Buffer (22336242), LanJieKe Technology Co., Ltd.; Reactive Oxygen Species Detection Kit (111922230307), Beyotime Biotechnology Co., Ltd.; Rhodamine-Phalloidin Staining Kit (237T0201), Shanghai Biosai Biotechnology Co., Ltd.; Anti-Fluorescence Quencher (ATWE09101), Yacoin Biotechnology Co., Ltd.

[0030] LRH-150 Biochemical Incubator, Electrothermal Constant Temperature Water Bath (HWS-24), Electrothermal Blast Drying Oven (DHG-9140A), Shanghai Hengke Scientific Instruments Co., Ltd.; SZ650 Stereo Microscope (Chongqing Optoelec Instrument Co., Ltd.); SW-CJ-2FD Clean Bench (Suzhou Antai Air Technology Co., Ltd.); ME104E Precision Electronic Balance (Mettler Toledo Instruments (Shanghai) Co., Ltd.); Primo R Desktop High-Speed Refrigerated Centrifuge, Smart 2Pure UV / VF Ultra-Pure Water Machine, VariosKanFlash Multifunctional Microplate Reader, Forma 900Series -80°C Ultra-Low Temperature Freezer, (Thermo Fisher Scientific Inc., USA); IMS-40 Automatic Snow Ice Maker (Changshu Xueke Electric Appliance Co., Ltd.); MQD-M1R Shaking Incubator (Shanghai Minquan Instruments Co., Ltd.); EBM2100VA-R 4 / -20°C Refrigerator (Electrolux (China) Electric Appliance Co., Ltd.); SX-500 Autoclave (TOMY Co., Japan); ST60-4 Microplate Thermostatic Oscillator (Hangzhou Mio Instruments Co., Ltd.); Axio Scope.A1 Inverted Fluorescence Microscope (Carl Zeiss AG, USA); Lecia TCS SP8 Laser Scanning Confocal Microscope (Leica Microsystems GmbH, Germany); KZ-III-FP High-Speed Low-Temperature Tissue Grinder (Wuhan Sevier Biotechnology Co., Ltd.); Fluorescent Quantitative PCR Instrument.

[0031] 1.2 Animals

[0032] Wild-type Caenorhabditis elegans N2 (hereinafter referred to as C. elegans) and Escherichia coli OP50 uracil synthesis-deficient strain are preserved by the Key Laboratory of Dai and Yi Medicines of Yunnan Province.

[0033] 2. Experimental Methods

[0034] 2.1 Preparation of NGM Plates for C. elegans Cultivation

[0035] Prepare NGM plates according to Table 1 (except for the egg-laying experiment, filter-sterilized Fudr should be added to a final concentration of 100 μM when preparing plates for other experiments), pour them onto plates of different diameters, and after solidification, let the water droplets on the plate wall dry at room temperature before use.

[0036] Table 1 Recipe for NGM Plates for C. elegans

[0037]

[0038] 2.2 Recovery of N2 C. elegans

[0039] Take out the cryopreserved nematode strain from the -80°C refrigerator. After natural thawing, discard the supernatant. Use a pipette to transfer the nematodes in the EP tube to a plate with OP50 bacteria, and place it in an incubator at 20°C with a relative humidity of 50% for cultivation.

[0040] 2.3 Nematode synchronization

[0041] Rinse the worms and eggs in the plate with deionized water into a 10 mL centrifuge tube. Add 3 mL of lysis solution (mix 5M NaOH solution and commercially available sodium hypochlorite solution evenly in a ratio of 1:2, and it can be used immediately after preparation). Shake well. When there are no nematode residues in the tube, centrifuge and discard the supernatant. Repeat this step 1 - 2 times. Resuspend with 10 mL of M9 and place it at 20°C with a relative humidity of 50%. The eggs will gradually hatch into larvae (L1 stage). Centrifuge and discard the supernatant. About 0.1 - 0.2 mL of M9 remains in the tube. Use a pipette to transfer the nematodes in the tube to a plate containing OP50 bacteria, and place it in an incubator at 20°C with a relative humidity of 50% for cultivation, then nematode synchronization can be achieved.

[0042] 2.4 Screening for the optimal effective concentration of the drug

[0043] 2.4.1 Preparation of Desmodium renalatum extract

[0044] Desmodium renalatum is extracted continuously by refluxing with 8 times the amount of 95% ethanol for 45 minutes. Repeat 3 times and then combine and filter by suction. Use a rotary evaporator to evaporate all the ethanol to obtain Desmodium renalatum extract (the extraction rate is 7%), and store it refrigerated for later use.

[0045] 2.4.2 Modeling method

[0046] Synchronize N2 nematodes at the L4 stage and place them on a culture dish containing 100 μM Fudr and 30 μM dexamethasone (DXMS) for 36 to 48 hours. (Fudr is not added to the culture dish for the egg-laying experiment)

[0047] 2.4.3 Group setting

[0048] Weigh the Desmodium renalatum extract and dissolve it in sterile water to a dosing concentration of 10 mg / ml (calculated according to the extraction rate of 7%, each ml contains 142.86 mg of crude drug) (add 0.2% Tween 80 to assist dissolution) as the stock solution, and use OP50 bacterial solution (OD 600=(0.6), and different concentrations of the drug administration groups (DXMS + Desmodium renalatum extract) were prepared with the mother liquor. The concentration gradients were: 8 mg / ml, 4 mg / ml, 2 mg / ml, 1 mg / ml, 0.5 mg / ml, 0.25 mg / ml, 0.125 mg / ml. The blank group (control group, without modeling and without drug administration) and the model group (DXMS group, with modeling and without drug administration) were added with the same volume of OP50 bacterial solution as the drug administration groups. After determining the optimal effective concentration of the drug through swimming and crawling experiments, the optimal concentration was used in subsequent experiments.

[0049] 2.4.4 Pre-culture

[0050] After synchronizing the nematodes and culturing them normally until the L4 stage, the nematodes were picked into the NGM plates of each experimental group and cultured at 20 °C for 36 to 48 h.

[0051] 2.4.5 Swimming experiment

[0052] After the end of 2.4.4, observe the swimming frequency of the nematodes: Pick the nematodes by group onto the NGM dishes with only pure water added. After the nematodes adapt for 5 min, observe the swimming frequency of the nematodes under a microscope at 40 times magnification for 30 s. The swimming frequency is counted according to the "C"-shaped swing rule of its body. One count is recorded when the front part swings from the left to the right and then back to the left. Each group has 3 parallel plates, and 10 nematodes are randomly counted in each culture dish.

[0053] 2.4.6 Crawling experiment

[0054] After the end of 2.4.4, observe the crawling times of the nematodes: Pick the nematodes by group onto the NGM dishes without food. After the nematodes adapt for 5 min, observe the crawling times of the nematodes under a microscope at 40 times magnification for 30 s. One crawl is recorded when its body makes a complete sine movement. Each group has 3 parallel plates, and 10 nematodes are randomly counted in each culture dish.

[0055] 2.5 Head swing experiment

[0056] After synchronizing the nematodes and culturing them normally until the L4 stage, the nematodes were picked into the NGM plates of each experimental group and cultured at 20 °C for 36 to 48 h. After the end, observe the head swing times of the nematodes: Pick the nematodes by group onto the NGM dishes without food. After the nematodes adapt for 5 min, observe and record the head swing times of the nematodes under a microscope at 40 times magnification for 30 s. One count is made when the head of the nematode taps lightly once. Each group has 3 parallel plates, and 10 nematodes are randomly counted in each culture dish.

[0057] 2.6 Pharyngeal pumping experiment

[0058] After synchronizing the nematodes and culturing them normally until the L4 stage, pick the nematodes into the NGM plates of each experimental group and culture them at 20 °C for 36 to 48 h. After that, observe the pharyngeal pumping frequency of the nematodes: Pick the nematodes into the NGM dishes without food according to the groups. After the nematodes adapt for 5 min, observe the frequency of the up and down shaking of the pharyngeal bulbs in the nematodes under a microscope at 100× magnification for 30 s. The shaking frequency is counted as one time when the pharyngeal bulb moves from top to bottom and then back to the top. Each group has 3 parallel plates, and 10 nematodes are randomly counted in each culture dish.

[0059] 2.7 Life-span experiment

[0060] After synchronizing the nematodes and culturing them normally until the L4 stage, transfer the nematodes to the medium with or without drugs (add Fudr that inhibits egg laying to each plate). Each group has 3 parallel plates, with a total of 90 - 150 nematodes, and culture them overnight in an incubator at 20 °C. This is recorded as the 0th day of the nematode survival. Use a stereomicroscope to observe the number of surviving nematodes in the medium every two days, and transfer the surviving nematodes to a new medium with the same drug concentration for continued culture. Record the survival of the nematodes until all of them die. When the nematodes show no response when stimulated with a picker and there is no transparent bacterial circle around the head, it is considered that the nematodes are dead. Exclude the nematodes that develop into bag-like worms and die outside the range coated with the experimental drug, and do not include them in the statistical data.

[0061] 2.8 Egg-laying experiment

[0062] After synchronizing the nematodes and culturing them normally until the L4 stage, pick the nematodes into the NGM plates containing 30 μM dexamethasone (the blank group does not contain dexamethasone). The inactivated OP50 contains 0, 0.5, 1, and 2 mg / ml Desmodium styracifolium extracts in turn. Randomly select 8 nematodes in each group and culture them at a constant temperature of 20 °C. Transfer the nematodes to the newly marked dishes for culture every day, and keep the old dishes for continued culture. On the third day, count the number of hatched worms in the old dishes on the second day. The experiment ends until the nematodes stop laying eggs. The total number of eggs laid by the same nematode every day is summed up to get the total number of eggs laid, and the number of eggs laid every day is counted as the daily average number of eggs laid.

[0063] 2.9 Rhodamine-phalloidin staining

[0064] After synchronizing the nematodes and culturing them to the L4 stage under normal conditions, pick the nematodes into the NGM plates of each experimental group and culture them at 20 °C for 36 to 48 h. Wash the nematodes from each plate with PBS and transfer them to 1.5 ml EP tubes. Wash the nematodes 3 times to remove Escherichia coli. Fix the nematodes on ice for 15 min with a PBS solution containing 3.75% formaldehyde. (Note: The preferred fixative is formaldehyde without methanol) Then wash 3 times with PBS. Add a PBS solution containing 0.5% Triton X-100 to permeabilize the nematodes at room temperature for 10 min and then wash 3 times with PBS. After discarding the supernatant, leave 0.1 - 0.2 ml of the nematode solution in the tube. Add 203 μL of phalloidin staining solution to each tube (3 μL of fluorescently labeled phalloidin stock solution diluted with 200 μL of PBS), incubate at room temperature for 20 min for staining. Finally, wash 2 - 3 times with PBS, add an anti-fluorescence quencher, and observe under a laser confocal microscope.

[0065] 2.10 Data processing

[0066] The above experiments were all independently repeated 3 times, and the experimental results were expressed as mean ± standard deviation (x ± s). The t-test or one-way analysis of variance (One-way ANOVA) was used to analyze the differences between groups, and the log-rank test was used for survival analysis. All experimental data were statistically analyzed and graphed using GraphPad Prism 8.0. *P < 0.05, **P < 0.01, ***P < 0.001 all indicated statistically significant differences.

[0067] 3. Experimental results

[0068] 3.1 Determination of the optimal effective concentration of the drug

[0069] Using Caenorhabditis elegans as the research object for muscle injury, first, a muscle injury model based on Caenorhabditis elegans needs to be established. By consulting the literature and combining the laboratory's research on related topics, this experiment used dexamethasone as the substance for muscle injury modeling, with a modeling dose of 30 μM and a modeling time of 36 - 48 h. In the normal activity state of Caenorhabditis elegans, there are some specific movement behavior patterns that use different parts of the muscles. Therefore, when constructing the muscle injury model, different movement patterns of the nematodes were selected to characterize the effect of the model. And during the activity of the nematodes, the body wall muscles are mainly used. At the same time, the body wall muscles are also the largest muscle part in the nematode body. So, the movement mode of the body wall muscles was used for experimental exploration.

[0070] Swimming and crawling are two ways of movement of nematodes in different states, and they have different movement characteristics. On solid plates, nematodes often move forward by crawling, that is, bending their bodies. This is mainly a medium-frequency, medium-amplitude movement. Compared with swimming, the muscles used in crawling are more biased towards the upper part of the body (head and middle of the body), and the tail muscles are used less. In liquid culture media, nematodes mainly move forward in the liquid by swimming and swinging their bodies. Swimming is a high-frequency, low-amplitude movement, and the tail muscles contribute more to the movement than during crawling. Therefore, the two movement experiments are combined to screen the optimal effective concentration of the drug. Figure 1 and Figure 2 As shown, within 30s, compared with the blank group (swimming: 44.57±4.99 times; crawling: 9.30±0.99 times), the swimming (39.07±4.52 times) and crawling (6.27±1.08 times) frequencies of the model group were reduced (***P<0.001), and the model was relatively successful; compared with the model group, the swimming and crawling frequencies of nematodes on the three culture dishes of 2, 1 and 0.5 mg / ml in the drug administration group were increased (***P<0.001), the swimming frequency increased by 13.65%, 14.59% and 17.58%, and the crawling frequency increased by 42.55%, 38.83% and 31.91%, respectively. Except for 8 mg / ml, which had a slightly better improvement effect on the crawling frequency of nematodes, the other concentrations were not as good as the above three concentrations. In summary, 2, 1 and 0.5 mg / ml were selected as the subsequent experimental drug administration groups.

[0071] 3.2 Effects of drugs on head shaking and pharyngeal pumping in nematodes

[0072] The muscle condition of nematodes can be represented by the nematode's locomotion ability. Therefore, changes in locomotion ability can often be used to characterize nematode muscle abnormalities or damage. In order to determine the effect of the extract of Delicata nematodes on muscle damage, the locomotion ability of different parts of nematodes was used in the experiment to explore.

[0073] Nematodes use head wiggling to find food and explore the surrounding environment. Head wiggling is the control of small muscles in the nematode's head. Figure 3 As shown, after dexamethasone injury, the average number of head swings of Caenorhabditis elegans decreased from 26.57±2.10 times in the blank group to 17.83±2.35 times. After protection by the 2, 1, and 0.5 mg / ml administration groups of the extract of Dexiella nephridia, the number of head swings increased by 20.37%, 38.50%, and 12.15%, respectively. Similarly, it can be intuitively seen that the number of head swings of nematodes decreased after dexamethasone injury, and recovered to a certain extent after protection by the extract of Dexiella nephridia (***P<0.001).

[0074] The pharynx is considered to be the neuromuscular organ of nematodes, and the muscle state of the pharynx can usually be judged by the high or low frequency of the pharyngeal pump (regular stretching and contraction movements of the pharynx). As Figure 4 , the pharyngeal pump rates of nematodes in five groups were measured in the experiment. The average pharyngeal pump frequencies within 30 s in the five groups were 79.87±4.12 times, 65.30±5.74 times, 71.70±6.89 times, 75.40±4.90 times, and 68.20±6.07 times, respectively. It can be clearly seen from the above experimental data that after dexamethasone injury, the pharyngeal pump frequency of nematodes decreased significantly. After intervention with Desmodium styracifolium extracts at 2, 1, and 0.5 mg / ml, the pharyngeal pump frequencies increased by 9.80%, 15.47%, and 4.44%, respectively. Except for the low concentration of 0.5 mg / ml, the improvement of the pharyngeal pump movement ability of nematodes at the other concentrations was more obvious (***P<0.001), indicating that Desmodium styracifolium extracts had a good protective effect on the pharyngeal pump ability of Caenorhabditis elegans after dexamethasone injury.

[0075] 3.3 Effects of drugs on the lifespan of nematodes

[0076] The length of the lifespan of nematodes is one of the indicators for evaluating the physiological state of nematodes, and it is carried out at 20 °C and a relative humidity of 50%. By evaluating the length of the lifespan of nematodes, the effects of dexamethasone on the survival of nematodes and the protective effect of Desmodium styracifolium extracts on it can be known. As Figure 5 shown, compared with the blank group, the survival percentage of nematodes in the model group decreased (***P<0.001); compared with the model group, Desmodium styracifolium extracts at 2 mg / ml and 1 mg / ml increased the survival rate of nematodes, which was statistically significant (*P<0.05). At the same time, Table 2 shows that compared with the model group, the average lifespan extension rates of each drug administration group were 8.53%, 10.68%, and 5.30% in turn, indicating that Desmodium styracifolium extracts helped to extend the overall lifespan of nematodes while improving the muscle injury of nematodes.

[0077] Table 2 Statistical table of nematode lifespan experiment (x±s, n = 3)

[0078]

[0079] Note: Compared with the model group, **P<0.01.

[0080] 3.4 Effects of drugs on the egg-laying ability of Caenorhabditis elegans

[0081] The number of eggs laid by nematodes is one of the indicators for evaluating the physiological state of nematodes, reflecting whether the drug has reproductive toxicity to nematodes. At the same time, the decline of reproductive function is directly correlated with lifespan aging. By evaluating the number of eggs laid by nematodes, we can know the effect of dexamethasone on the reproductive ability of nematodes and the protective effect of Desmodium renifolium extract on it. The experimental results showed that in terms of the number of eggs laid, the average number of eggs laid in each group was 163.88±14.32, 159.88±17.18, 162.38±20.60, 168.50±21.42 and 169.50±18.42 respectively. There were slight differences in the data. However, due to the large individual differences of nematodes within each group, there was no significant difference between groups after data analysis (P>0.05), indicating that dexamethasone had no effect on the total number of eggs laid by nematodes (see Figure 6 ).

[0082] 3.5 Rhodamine-phalloidin staining

[0083] The body wall muscles of nematodes were stained with rhodamine-phalloidin. The experimental results are as Figure 7 shown. Compared with the blank group, after being damaged by dexamethasone in the model group, the muscle stripes of the nematode body wall showed fracture and fault phenomena. After protection with Desmodium renifolium extract, this situation was greatly improved. The muscle stripes of the nematode body wall in each administration group were clear and regularly arranged. Although there were occasional wavy myofilaments, the staining patterns at the three protective concentrations of 2, 1 and 0.5 mg / ml were not much different from those of the blank group.

[0084] In summary, combining the indicators in the part of nematode locomotor ability, it can be concluded from this experiment that using dexamethasone to construct a muscle injury model for N2 nematodes will reduce the overall locomotor ability of nematodes, and it can also be found through staining that the body wall muscles are damaged. Protection with Desmodium renifolium extract can reverse the decline of locomotor function, and at the same time, the body wall muscles can be restored to a certain extent, and the overall lifespan of nematodes can be extended.

Claims

1. Use of the ethanol extract of Desmodium renifolium in the preparation of a medicament for treating sarcopenia.

2. The use according to claim 1, wherein: the ethanol extract of Desmodium renifolium is a 95% ethanol extract; the 95% ethanol extract is prepared by refluxing Desmodium renifolium with 95% ethanol and removing the ethanol from the extract.

3. The use according to claim 1, wherein: the medicament is a medicament for improving the decline in motor function.

4. The use according to claim 3, wherein: the medicament is a medicament for improving the decline in motor function caused by muscle injury.

5. The use according to claim 1, wherein: the medicament is a medicament for improving the shortening of lifespan caused by the decline in motor ability.

6. The use according to claim 1, wherein: the medicament is a medicament for repairing damaged muscles.

7. The use according to claim 1, wherein: the medicament is a preparation prepared by adding a pharmaceutically acceptable excipient to the ethanol extract of Desmodium renifolium as an active ingredient.

8. The use according to claim 7, wherein: the preparation is an oral preparation, an external preparation or an injectable preparation.

9. The use according to claim 8, wherein: the oral preparation is a granule, a solution, a pill, a capsule or a powder; the external preparation is a plaster, a spray, a liniment or an emulsion; the injectable preparation is an injectable solution or a powder for injection.

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