CN113304179B

CN113304179B - Application of whole plant extract of pachyrhizus in preparation of medicines for supplementing calcium and preventing osteoporosis

Info

Publication number: CN113304179B
Application number: CN202110750765.7A
Authority: CN
Inventors: 刘红梅
Original assignee: Ningxia Milk Melon Biotechnology Co ltd
Current assignee: Ningxia Milk Melon Biotechnology Co ltd
Priority date: 2021-07-02
Filing date: 2021-07-02
Publication date: 2022-08-12
Anticipated expiration: 2041-07-02
Also published as: CN113304179A

Abstract

The invention discloses an application of pachyrhizus in preparing medicines for supplementing calcium and preventing osteoporosis; the sweet potato medicinal material powder and extract can improve the serum calcium concentration of rats, the content of 1, 25-dihydroxy vitamin D3(DHVD3) and the content of Calcitonin (CT), and reduce the content of alkaline phosphatase and parathyroid hormone (PTH) of rats, and pass the indexes. The application can be used for promoting calcium absorption and calcium transfer from serum to bone and preventing osteoporosis by supplementing serum calcium.

Description

Application of whole plant extract of pachyrhizus in preparation of medicines for supplementing calcium and preventing osteoporosis

Technical Field

The invention relates to the technical field of phytochemistry and biological pharmacy, in particular to application of pachyrhizus in preparing medicines for supplementing calcium and preventing osteoporosis.

Background

At present, calcium carbonate is mainly used as a calcium supplement, but when the calcium carbonate is taken for a long time, eructation and constipation easily occur, occasionally, choline syndrome is generated, the symptoms of hypercalcemia, alkalosis and renal insufficiency occur, and when the calcium carbonate is taken for a long time, the gastric acid secretion rebound is increased, and hypercalcemia can occur. In addition, osteoporosis and calcium deficiency are common diseases of the elderly, gastrointestinal tract function of the elderly is weakened, side effects are more likely to occur when calcium carbonate is taken, and the search for novel calcium supplement preparations is urgent.

The radix Pachyrhizi Erosi is a perennial herb of Annonaceae, and has effects of invigorating lung qi, clearing heat, lowering fire, promoting fluid production, quenching thirst, relieving inflammation, relieving pain, dispelling pathogenic wind, promoting blood circulation, and relieving diarrhea by adding traditional Chinese medicines into whole herb and fruit, and can be used for treating enteronitis, dysentery, galactostasis, etc. by topical application. In recent years, studies on the sweet potatoes mainly focus on chemical components, tissue culture and artificial domestication, content measurement and quality standards, and the studies on practical products are less. To date, the effect of the sweet potato on calcium supplement and osteoporosis resistance has not been found in the field.

Therefore, how to apply the radix cynanchi bungei to the preparation of the medicines for supplementing calcium and preventing osteoporosis is a problem which needs to be solved urgently by the technical personnel in the field.

Disclosure of Invention

In view of the above, the invention provides an application of radix cynanchi bungei in preparing a medicine for supplementing calcium and preventing osteoporosis.

In order to achieve the purpose, the invention adopts the following technical scheme:

application of radix Pachyrhizi Erosi in preparing medicine for supplementing calcium and preventing osteoporosis is provided.

As a preferred technical scheme of the invention, the pachyrhizus can improve the calcium content in serum.

As a preferred technical scheme of the invention, the pachyrhizus can reduce the content of alkaline phosphatase in serum.

As a preferred technical scheme of the invention, the Dioscorea nipponica can improve the content of DHVD3 in serum.

As a preferred technical scheme of the invention, Dixie can reduce the content of parathyroid hormone in serum.

As a preferred technical scheme of the invention, the pachyrhizus can improve the content of calcitonin in serum.

According to the technical scheme, compared with the prior art, the invention discloses the application of the pachyrhizus in preparing the medicines for supplementing calcium and preventing osteoporosis, the pachyrhizus is used for a long time in the field of traditional Chinese medicines, the dosage is large, and adverse reactions are not reported. Experimental results show that the sweet potato medicinal material powder and extract can improve the serum calcium concentration of rats, the content of 1, 25-dihydroxy vitamin D3(DHVD3) and the content of Calcitonin (CT), and reduce the content of alkaline phosphatase and parathyroid hormone (PTH) of rats, and the indexes are passed through. The application can be used for promoting calcium absorption and calcium transfer from serum to bone and preventing osteoporosis by supplementing serum calcium.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.

FIG. 1 is a graph showing the change of calcium content in serum of Pachyrhizus erosus in different treatment groups;

FIG. 2 is a graph showing the change in serum alkaline phosphatase levels in different groups of Pachyrhizus;

FIG. 3 is a graph showing the change of DHVD3 content in blood serum of different groups of Pachyrhizus;

FIG. 4 is a graph showing the change of parathyroid hormone content in blood serum of different groups of Pachyrhizus;

FIG. 5 is a graph showing the change of calcitonin content in blood serum of different groups of Pachyrhizus Erecta.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The apparatus used in the examples:

calcium (Ca) colorimetric test kit (manufacturer: Elapscience cat. No.: E-BC-K103-M);

alkaline phosphatase (ALP) colorimetric test kit (manufacturer: Elapscience cat # E-BC-K091-M);

1,25 dihydroxy vitamin D3(DHVD3) enzyme-linked immunosorbent assay kit (manufacturer: Elabccience, cat # E-EL-0016 c);

rat complete parathyroid hormone (I-PHT) enzyme-linked immunosorbent assay kit (manufacturer: Elabccience, cat # E-EL-R0535 c);

rat Calcitonin (CT) ELISA kit (manufacturer: Elapscience, cat # E-EL-R0047 c).

Example 1 preparation of Whole plant powder and fruit powder of sweet potato

Weighing 500g of whole sweet potato, cutting into sections of 0.5cm, pulverizing with a traditional Chinese medicine pulverizer, and sieving with a traditional Chinese medicine sieve of 80 meshes to obtain the whole sweet potato powder. Dried fruits of the sweet potato are 500g, cut in half, crushed by a traditional Chinese medicine crusher, and sieved by a traditional Chinese medicine sieve of 80 meshes to obtain the sweet potato powder.

Example 2 preparation of extract of whole plant and fruit of Japanese pachyrhizus

Taking whole sweet potato, cutting into 0.5cm sections, weighing 300g, placing into a 5L round-bottom flask, adding 3000mL of distilled water, soaking for 30 minutes, heating and refluxing for 1 hour by adopting an electric heating jacket (the extraction temperature is 100 ℃, filtering by using 8 layers of gauze, combining filtrate repeatedly for three times, pouring the filtrate into a 250mL evaporating dish in portions, evaporating and drying on a water bath pan (the temperature of the water bath pan is 90 ℃), pouring the filtrate again when the filtrate in the evaporating dish is quick-dried until all the filtrate is evaporated to dryness, drying until all the filtrate becomes crisp by pressing, weighing 100.8g of extract, controlling the yield to be 33-34% by the method, and repeatedly preparing about 1kg of extract for later use.

Taking 250g of dried pachyrhizus fruits, cutting the dried fruits into half parts, putting the cut fruits into a 5L round-bottom flask, adding 2500mL of distilled water, soaking for 30 minutes, heating and refluxing for 1 hour by adopting an electric heating sleeve (the extraction temperature is 100 ℃), filtering by using 8 layers of gauze, repeating the three times, combining the filtrate, pouring the filtrate into a 250mL evaporation dish in batches, putting the evaporation dish on a water bath, evaporating and drying (the temperature of the water bath is 90 ℃), and pouring the filtrate again when the filtrate in the evaporation dish is quick-dried until all the filtrate is evaporated to dryness. Drying until it becomes brittle, and weighing 103.8g of extract. According to the method, the yield is 34-35%, and about 1kg of extract needs to be prepared repeatedly for later use.

EXAMPLE 3 establishment and application of calcium deficiency animal model

Selecting 80 healthy rats with the weight of 200 +/-20 g, randomly extracting 8 rats as a normal group, feeding a maintenance feed (the calcium content of the normal maintenance feed is 1.1%), and performing modeling treatment on 72 rats, feeding a low-calcium purified feed (the calcium content is 0.01%, the feed formula comprises 20% of casein, 38.81% of corn starch, 13.2% of maltodextrin, 10% of sucrose, 5% of cellulose, 7% of soybean oil, 5.99% of calcium-deficient minerals and other small materials, wherein the feed is purchased from Jiangsu Dison biomedicine Limited company, and drinking deionized water to avoid obtaining calcium from drinking water; after six weeks of administration, the serum of rats in different groups in the serum of rats in the model group and the normal group is measured and determined to have the contents of calcium, alkaline phosphatase (ALP), 1, 25-dihydroxy vitamin D3(DHVD3), parathyroid hormone (PTH) and Calcitonin (CT), and the result shows that the model group and the normal group have obvious difference and the modeling is successful; then randomly dividing rats in the model group into 9 groups, wherein each group comprises 8 rats, and respectively dividing the rats into a positive medicine group, a pachyrhizus fruit extract group with low dose, a medium dose and a high dose group, and a pachyrhizus whole-grass extract group with low dose, a medium dose and a high dose group;

the groups were treated as follows:

a positive drug group: a calcium supplement product (Langdi calcium carbonate D3 granules) sold on the market by intragastric administration is administered according to the administration amount recommended by a positive medicine instruction and according to the proportion of human and mouse (1: 6) (calculated by calcium: 50 mg/kg);

high dose group of pachyrhizus extract: performing intragastric administration treatment on the trichosanthes kirilowii fruit extract prepared in the example 2, wherein the intragastric administration amount is 7.2 g/kg;

the dosage group of the pachyrhizus extract is as follows: performing intragastric administration treatment on the trichosanthes kirilowii fruit extract prepared in the example 2, wherein the intragastric administration amount is 3.6 g/kg;

low dose group of pachyrhizus extract: performing intragastric administration treatment on the trichosanthes kirilowii fruit extract prepared in the example 2, wherein the intragastric administration amount is 1.8 g/kg;

and (3) ground-soft-sweet fruit powder group: performing intragastric administration treatment on the mirabilis jalapa fruit powder prepared in the example 1, wherein the intragastric administration amount is 0.46 g/kg;

the high-dose group of the whole-grass extract of the pachyrhizus comprises: performing intragastric administration treatment on the whole plant extract of the pachyrhizus prepared in example 2, wherein the intragastric administration amount is 7.2 g/kg;

the dose groups of the whole-grass extract of the pachyrhizus are as follows: performing intragastric administration treatment on the whole plant extract of the pachyrhizus prepared in the example 2, wherein the intragastric administration amount is 3.6 g/kg;

the low dose group of the whole grass extract of pachyrhizus: performing intragastric administration treatment on the whole plant extract of the pachyrhizus prepared in the example 2, wherein the intragastric administration amount is 1.8 g/kg;

and (3) sweet potato grass powder group: performing intragastric administration treatment on the cynanchum wilfordii powder prepared in the example 1, wherein the intragastric administration amount is 1.1 g/kg;

dosing was continued for 6 weeks. After 6 weeks, blood was taken from the abdominal aorta, centrifuged at 3000r/min for 10 minutes, and the upper serum was taken. The contents of calcium, alkaline phosphatase (ALP), 1, 25-dihydroxy vitamin D3(DHVD3), parathyroid hormone (PTH) and Calcitonin (CT) in the blood serum of rats in different groups are measured by a calcium (Ca) colorimetric test box, an alkaline phosphatase (ALP) colorimetric test box, a 1, 25-dihydroxy vitamin D3(DHVD3) enzyme-linked immunosorbent assay kit, a rat whole-segment parathyroid hormone (I-PHT) enzyme-linked immunosorbent assay kit and a rat Calcitonin (CT) enzyme-linked immunosorbent assay kit, and the therapeutic effects of different administration groups on osteoporosis are compared.

Calcium is the most abundant cation in the body and bone is the largest reserve of calcium in the body. The calcium in the serum and the calcium in the bone are maintained in a dynamic balance, and when the bone is in calcium deficiency, the calcium deficiency can be expressed by the calcium content in the serum. Serum calcium is an important index for evaluating whether the body is calcium-deficient.

The bone-derived alkaline phosphatase (ALP) is secreted from bone, and when the calcium salt precipitation in bone is insufficient, the enzyme secretion is increased, and when the calcium salt in bone is sufficient, the secretion is reduced. Therefore, the lack of calcium in bone can be reflected by alkaline phosphatase.

1, 25-dihydroxy vitamin D3(DHVD3) can promote the absorption of calcium and phosphorus by small intestine mucosa, increase the absorption capacity of calcium, and improve the utilization rate of calcium.

Parathyroid hormone (PTH) is a basic single-chain polypeptide hormone secreted by parathyroid chief cells. Its main function is to regulate the metabolism of calcium and phosphorus in vertebrates, and promote the increase of calcium level and decrease of phosphorus level in blood. PTH contributes to elevated plasma calcium concentrations and the major target organs of action are bone and kidney. It mobilizes bone calcium to enter blood, promotes reabsorption of calcium ions and excretion of phosphate by renal tubules, and increases blood calcium concentration.

Calcitonin (CT) is a polypeptide hormone involved in the metabolism of calcium-containing bone, has acute inhibitory effect on osteoclast, and can reduce the migration of calcium from bone to blood. Calcitonin can also inhibit the dissolution and transfer of bone salt, inhibit the decomposition of bone matrix, and increase the bone renewal rate.

Therefore, the change of five indexes in the serum of the rat is measured, and the effect of calcium supplement and osteoporosis prevention of the snake gourd can be powerfully proved; the adopted method is operated according to the kit operation instructions, an enzyme labeling instrument is adopted to measure OD values, the measured index concentration in each sample is calculated according to a standard curve, and the influence of different dosing of the pachyrhizus on different indexes in serum is evaluated; the results are shown in table 1, fig. 2, fig. 3, fig. 4 and fig. 5;

TABLE 1 Effect of sweet potato on five factors in serum

Note: compared with the model group, P is less than 0.05, P is less than 0.01, the fruit powder group is the sweet potato fruit powder group, the grass powder group is the sweet potato whole grass powder group, the fruit high, medium and low dosage groups are the sweet potato fruit extract high, medium and low dosage groups, and the grass high, medium and low dosage groups are the sweet potato whole grass extract high, medium and low dosage groups. The positive medicine is a calcium supplement product (Langdi calcium carbonate D) sold on the market ₃ Particles).

As can be seen from FIG. 1, the calcium deficiency model has a very significant difference (P < 0.01) compared with the normal group, indicating that the calcium deficiency model is successfully molded. Compared with a model group, the sweet potato fruit powder, the grass powder and the extract have the effect of improving the concentration of calcium in serum, and have very obvious difference. The whole grass, fruit and extract of the sweet potato have the effect of supplementing calcium.

As can be seen from FIG. 2, the calcium deficiency model has a very significant difference (P < 0.01) compared with the normal group, indicating that the calcium deficiency model is successful. Compared with the model group, the high and medium doses of the whole sweet potato grass have very significant difference (P is less than 0.01), and the low dose of the whole sweet potato grass has no significant difference (P is more than 0.05) compared with the model group. The high, medium and low dose doses of the sweet potato fruits have very significant difference (P is less than 0.01) compared with the model group. The sweet potato powder has significant difference (P is less than 0.05) compared with the model group. The cynanchum kochianum powder has very significant difference (P < 0.01) compared with the model group. The contents of calcium salt in bone substance and osteoporosis are improved.

As can be seen from FIG. 3, the calcium deficiency model has significant difference (P < 0.05) compared with the normal group, indicating that the calcium deficiency model is successful. Compared with the model group, the high and medium doses of the whole sweet potato grass have significant difference (P is less than 0.01), and the low dose of the whole sweet potato grass has significant difference (P is less than 0.05) compared with the model group. The high and medium doses of the sweet potato fruits have very significant difference (P is less than 0.01) compared with the model group. The low dose of the pachyrhizus has significant difference compared with the model group (P < 0.05). The sweet potato can promote the absorption and utilization of calcium by the body.

As can be seen from FIG. 4, the calcium deficiency model has significant difference (P < 0.05) compared with the normal group, indicating that the calcium deficiency model is successful. Compared with the model group, the high and medium doses of the whole sweet potato grass have significant difference (P is less than 0.01), and the low dose of the whole sweet potato grass has significant difference (P is less than 0.05) compared with the model group. The high and medium doses of the sweet potato fruits have very significant difference (P is less than 0.05) compared with the model group. The low dose of the pachyrhizus has significant difference compared with the model group (P < 0.05). The increase of parathyroid hormone indicates that the body is lack of calcium and needs to convert bone calcium into blood, and experimental results show that the pachyrhizus can increase the calcium concentration in blood, reduce the parathyroid hormone content, inhibit the conversion of bone calcium into blood calcium and inhibit osteoporosis.

As can be seen from FIG. 5, the calcium deficiency model has significant difference (P < 0.05) compared with the normal group, indicating that the calcium deficiency model is successful. Compared with the model group, the high and medium doses of the whole sweet potato grass have significant difference (P is less than 0.01), and the low dose of the whole sweet potato grass has significant difference (P is less than 0.05) compared with the model group. The high doses of the sweet potato fruits have very significant differences (P < 0.01) compared with the model group. The medium and low doses of the pachyrhizus have significant difference compared with the model group (P < 0.05). The sweet potato fruit powder has very significant difference (P < 0.01) compared with the model group. The experimental result shows that the pachyrhizus can improve the content of calcitonin, promote the transfer of calcium in serum to bone calcium, improve bone quality and prevent osteoporosis.

The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other.

The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. The application of the whole plant extract of the mirabilis jalapa in preparing medicines for supplementing calcium and preventing osteoporosis; the whole plant extract of the cynanchum glaucescens can improve the calcium content in serum, reduce the alkaline phosphatase content in serum, improve the 1, 25-dihydroxyvitamin D3 content in serum, reduce the parathyroid hormone content in serum and improve the calcitonin content in serum.