CN113908235A

CN113908235A - Traditional Chinese medicine compound preparation for treating diabetic retinopathy and preparation method and application thereof

Info

Publication number: CN113908235A
Application number: CN202111459288.5A
Authority: CN
Inventors: 赵越; 周希乔; 余江毅; 安晓飞; 余旭; 吴豪; 娄妍; 周正国; 王恒; 沈建江
Original assignee: Jiangsu Provincial Hospital of Chinese Medicine
Current assignee: Jiangsu Provincial Hospital of Chinese Medicine
Priority date: 2021-12-02
Filing date: 2021-12-02
Publication date: 2022-01-11
Anticipated expiration: 2041-12-02
Also published as: CN113908235B

Abstract

The invention relates to a traditional Chinese medicine compound preparation for treating diabetic retinopathy, a preparation method and application thereof. The traditional Chinese medicine compound preparation is prepared from the following traditional Chinese medicines: radix astragali, radix rehmanniae, herba Dendrobii, flos Abelmoschi Manihot, cortex moutan, radix Salviae Miltiorrhizae, radix Notoginseng powder, and flos Buddlejae. The traditional Chinese medicine compound preparation can be used for treating diabetic retinopathy, and the molecular level of the traditional Chinese medicine compound preparation is determined to be the over-expression of TGF-beta 1 protein and mRNA, VEGF protein and mRNA in retinal tissues under diabetic retinopathy, and the miR-200b level in the retinal tissues under diabetic retinopathy is promoted to be improved, so that the specific conditions of diseases can be accurately matched, and the diagnosis and treatment effects are favorably improved.

Description

Traditional Chinese medicine compound preparation for treating diabetic retinopathy and preparation method and application thereof

Technical Field

The invention relates to a traditional Chinese medicine compound preparation for treating diabetic retinopathy, a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicines.

Background

To the best of the inventor's knowledge, diabetic retinopathy (hereinafter referred to as DR) is one of the major microvascular complications of diabetes (hereinafter referred to as DM), which is called "diabetes eye disease" in traditional Chinese medicine. Recent meta-analysis showed: the total incidence of DR in the DM population is about 34.6%, and DR has become the leading cause of vision loss in middle aged and elderly people worldwide. At present, the pathogenesis of DR is not completely clarified, and an effective intervention means is actively searched for, which has extremely important significance for preventing and treating DR.

In the latest DR clinical prevention and treatment guidelines established by the American ophthalmologic Association, strict control of metabolic disorders such as blood sugar, blood pressure, and blood fat is still used as a main drug prevention and treatment means. And the whole retinal photocoagulation, anti-VEGF monoclonal antibody retrobulbar injection and vitrectomy are selectively used when severe nonproliferative diabetic retinopathy (NPDR) or Proliferative Diabetic Retinopathy (PDR) occurs and the corresponding indications are combined. It is also recommended in the guidelines that ophthalmologists with experience in clinical management of DR be timely referred to patients with Diabetic Macular Edema (DME), severe NPDR and PDR. Therefore, the selection of appropriate drugs to alleviate and prevent further exacerbation of the lesions in the early stages of DR is an important responsibility of all endocrinologists and general practitioners responsible for DR management.

Regarding the pharmaceutical intervention of DR, in addition to basic treatments such as controlling blood sugar, blood pressure, blood lipid, etc., other drugs such as microcirculation improving drugs, radical scavengers, antithrombotic drugs, diacylglycerol-protein kinase C inhibitors, etc., may have a certain therapeutic effect on DR, but their effects are relatively limited, and further intensive studies are required.

Traditional Chinese medicine accumulates abundant experience in long-term clinical practice in the formation and prevention of DR. The treatment means aiming at DR comprises various modes such as dialectical treatment, stage treatment, special medicines, traditional Chinese medicine extracts, acupuncture and moxibustion, traditional Chinese medicine ion introduction and the like. Relevant researches suggest that the traditional Chinese medicine can relieve the apoptosis of retinal pericytes and improve the functions of endothelial cells by improving oxidative stress and inflammatory states, and can also inhibit the proliferation of retinal neovessels by regulating certain cytokines such as hypoxia inducible factor-1 alpha, insulin growth factor-1, interleukin-6, tumor necrosis factor-alpha and the like so as to achieve the aim of improving DR. However, the research on DR intervention of the traditional Chinese medicine compound preparation is mainly based on clinical observation, and research evidence for animal experiments and related mechanism exploration is relatively few, so that research results in the aspect are urgently needed to be added.

The invention patent applications with application numbers CN20061007164.1 and CN1857531A disclose a Chinese medicine preparation prescription for treating diabetic retinopathy and a preparation method thereof through retrieval; the invention patents of patent No. CN201110194682.0 and publication No. CN102258709B also disclose a Chinese medicinal preparation for treating diabetic retinopathy and a preparation method thereof. The technical schemes are different from the research results related to the invention, and the technical schemes do not carry out relevant research on the aspect of mechanism exploration, so that the improvement space still exists.

Disclosure of Invention

The main purposes of the invention are: the traditional Chinese medicine compound preparation can inhibit the over-expression of TGF-beta 1 protein and mRNA, VEGF protein and mRNA in retinal tissues under the pathological changes, and can promote the increase of miR-200b level in the retinal tissues under the pathological changes. Meanwhile, also provides a preparation method and application of the corresponding Chinese medicinal compound preparation.

The technical scheme for solving the technical problems of the invention is as follows:

a traditional Chinese medicine compound preparation for treating diabetic retinopathy is characterized by being prepared from the following traditional Chinese medicines: radix astragali, radix rehmanniae, herba Dendrobii, flos Abelmoschi Manihot, cortex moutan, radix Salviae Miltiorrhizae, radix Notoginseng powder, and flos Buddlejae; the weight ratio of the astragalus root, the dried rehmannia root, the dendrobium, the flower of abelmoschus manihot, the cortex moutan, the salvia miltiorrhiza, the pseudo-ginseng powder and the pale butterflybush flower is 15-30: 10-20: 10-20: 30-60: 10-20: 15-30: 3-6: 10-15.

In the traditional Chinese medicine compound preparation: radix astragali, sweet in taste, slightly warm in nature, can tonify qi, promote diuresis and remove stagnation; dried rehmannia root, sweet in taste and cold in nature, can nourish yin and promote the production of body fluid; dendrobe, sweet in taste, slightly cold in nature, can nourish yin and promote the production of body fluid, and the three are combined to be a main drug for tonifying qi and yin; abelmoschus manihot is sweet in flavor and cold in nature, and has the effects of clearing away damp-heat; cortex moutan, bitter and pungent in flavor and slightly cold in nature, has the actions of cooling blood and promoting blood circulation; dan Shen is bitter in taste and slightly cold in nature, and has the actions of removing blood stasis and cooling blood; pseudo-ginseng, sweet in taste, slightly bitter in taste, warm in nature, has the functions of removing blood stasis and stopping bleeding, combines the four, and is a main drug for resolving dampness and activating blood circulation; flos Buddlejae is sweet in flavor and slightly cold in nature, and has the effects of clearing heat, nourishing liver and improving eyesight. The whole formula is combined with warm and cool, tonifies and clears, and has the effects of tonifying qi and yin, and eliminating dampness and promoting blood circulation.

Experiments prove that the traditional Chinese medicine compound preparation can improve the pathological change of retina, improve the condition of retinal capillary, inhibit the over-expression of TGF-beta 1 protein and mRNA, VEGF protein and mRNA in the retinal tissue and promote the increase of miR-200b level in the retinal tissue for the retinal tissue under diabetic retinopathy.

The technical scheme for further perfecting the traditional Chinese medicine compound preparation is as follows:

preferably, the weight ratio of the astragalus to the radix rehmanniae, the dendrobium to the abelmoschus manihot, the cortex moutan to the radix salviae miltiorrhizae to the radix notoginseng powder to the flos buddlejae is 15: 15: 10: 30: 15: 15: 3: 10.

preferably, the traditional Chinese medicine compound preparation is prepared from the following traditional Chinese medicines: 15-30 g of astragalus, 10-20 g of radix rehmanniae recen, 10-20 g of dendrobium, 30-60 g of abelmoschus manihot, 10-20 g of cortex moutan, 15-30 g of salvia miltiorrhiza, 3-6 g of pseudo-ginseng powder and 10-15 g of buddleja officinalis.

More preferably, the traditional Chinese medicine compound preparation is prepared from the following traditional Chinese medicines: 15g of astragalus, 15g of radix rehmanniae recen, 10g of dendrobium, 30g of flower of abelmoschus manihot, 15g of cortex moutan, 15g of salvia miltiorrhiza, 3g of pseudo-ginseng powder and 10g of buddleja officinalis.

By adopting the preferred scheme, the weight ratio of the traditional Chinese medicines can be further optimized.

The present invention also provides:

the preparation method of the traditional Chinese medicine compound preparation for treating diabetic retinopathy is characterized in that the traditional Chinese medicine compound preparation is a traditional Chinese medicine compound decoction; the preparation method comprises the following steps:

firstly, soaking astragalus, radix rehmanniae, dendrobium, abelmoschus manihot, cortex moutan, salvia miltiorrhiza and buddleja officinalis in clean water for 30-60 minutes according to the prescription amount, and then putting the soaked materials into a medicine decocting machine;

secondly, adding a predetermined amount of water into the medicine decocting machine, and decocting for 30-60 minutes; squeezing out the decoction by using a mechanical squeezing part of the decoction machine;

and thirdly, adding the pseudo-ginseng powder with the prescription amount into the medicine juice, uniformly mixing, and subpackaging the obtained liquid to obtain a finished product.

Preferably, in the second step, the predetermined amount is 500-600 ml of water corresponding to each dose of the Chinese herbal compound preparation.

The preparation method of the traditional Chinese medicine compound preparation for treating diabetic retinopathy is characterized in that the traditional Chinese medicine compound preparation is a traditional Chinese medicine compound extract; the preparation method comprises the following steps:

firstly, taking the astragalus, radix rehmanniae, dendrobium, abelmoschus manihot flower, cortex moutan, salvia miltiorrhiza and buddleja officinalis in the prescription amount, and decocting for at least 2 times with water, wherein each time is 1-2 hours; mixing the decoctions, and filtering to obtain filtrate;

secondly, concentrating the filtrate at 65-85 ℃ under reduced pressure for 3-4 hours to obtain extract; continuously stirring the extract at 80 +/-5 ℃ for 30-40 minutes, adding the formula amount of pseudo-ginseng powder, and continuously stirring until the mixture is uniformly mixed;

and thirdly, spray drying the obtained mixture to obtain a finished product.

By adopting the preparation method, the traditional Chinese medicine compound decoction or the traditional Chinese medicine compound extract can be prepared and obtained for patients to take.

Preferably, in the second step, the stirring time after the pseudo-ginseng powder is added is 5-10 minutes.

The present invention also provides:

the application of the traditional Chinese medicine compound preparation in preparing the medicine for treating diabetic retinopathy.

Preferably, the therapeutic effects of the medicament comprise improving pathological changes of retina under diabetic retinopathy, improving the condition of retinal capillaries under diabetic retinopathy, inhibiting over-expression of TGF-beta 1 protein and mRNA and VEGF protein and mRNA in retinal tissue under diabetic retinopathy and promoting the increase of miR-200b level in retinal tissue under diabetic retinopathy.

The traditional Chinese medicine compound preparation can be used for treating diabetic retinopathy, and the molecular level of the traditional Chinese medicine compound preparation is determined to be the over-expression of TGF-beta 1 protein and mRNA, VEGF protein and mRNA in retinal tissues under diabetic retinopathy, and the miR-200b level in the retinal tissues under diabetic retinopathy is promoted to be improved, so that the specific conditions of diseases can be accurately matched, and the diagnosis and treatment effects are favorably improved.

Drawings

FIG. 1 is a graph showing a comparison of HE staining results (HE X200) of retinal tissues of rats in each group according to the example of the present invention.

FIG. 2 is a graph comparing the results of retinal vascular digestion plating of rats in each group according to the example of the present invention (PAS X200).

FIG. 3 is a graph showing immunofluorescence comparison of TGF-beta 1 and VEGF proteins in rat retinal tissues in various groups according to an embodiment of the present invention.

FIG. 4 is a graph showing the comparison of the expression levels of TGF-beta 1 and VEGF proteins in rat retinal tissues in accordance with the present invention.

FIG. 5 is a graph showing the comparison of the expression of TGF-. beta.1 mRNA, miR-200b and VEGF mRNA in the retinal tissues of rats in each group according to the example of the present invention.

Detailed Description

In the research of the project, the study on the known formulas proves that the traditional Chinese medicines such as radix rehmanniae, cortex moutan and abelmoschus manihot have the curative effect on diabetic retinopathy (hereinafter referred to as DR), and provides a basis for treating DR by a 'yin nourishing, dampness resolving and blood circulation activating method'. Then, on the basis of literature research, by combining the pathogenesis characteristics of DR 'deficiency of both qi and yin and dampness and blood stasis and collateral obstruction' and the clinical experience of DR diagnosis and treatment in the past, the formula of the traditional Chinese medicine compound preparation is drawn up as follows: radix astragali, radix rehmanniae, herba Dendrobii, flos Abelmoschi Manihot, cortex moutan, radix Salviae Miltiorrhizae, radix Notoginseng powder, and flos Buddlejae. Then, the dosage of the Chinese medicament is subjected to a groping experiment, and the following steps are determined: the weight ratio of the astragalus root, the dried rehmannia root, the dendrobium, the flower of abelmoschus manihot, the root bark of the peony tree, the root of red-rooted salvia, the pseudo-ginseng powder and the pale butterflybush flower is 15-30: 10-20: 10-20: 30-60: 10-20: 15-30: 3-6: 10-15 parts of; in the experimental process, after the traditional Chinese medicine compound preparation (decoction) is taken by a patient with DR (digital radiography) combined macular edema, the vision is obviously improved, and the macular edema is reduced; however, some patients feel dry heat after taking the medicine, so the dosage of the astragalus is adjusted to be reduced to the range, the cortex moutan is adjusted to be increased to the range, and the rest patients do not change, so the dry heat symptom of the patients is improved. Finally, animal and clinical studies are carried out under the determined dosage ratio.

In specific implementation, the traditional Chinese medicine compound preparation for treating diabetic retinopathy is prepared from the following traditional Chinese medicines: radix astragali, radix rehmanniae, herba Dendrobii, flos Abelmoschi Manihot, cortex moutan, radix Salviae Miltiorrhizae, radix Notoginseng powder, and flos Buddlejae; wherein, the weight ratio of the astragalus root, the dried rehmannia root, the dendrobium, the flower of abelmoschus manihot, the root bark of the peony tree, the root of red-rooted salvia, the pseudo-ginseng powder and the pale butterflybush flower is 15-30: 10-20: 10-20: 30-60: 10-20: 15-30: 3-6: 10-15.

The above weight ratio is preferably: 15: 15: 10: 30: 15: 15: 3: 10.

the dosage composition is preferably: 15-30 g of astragalus, 10-20 g of radix rehmanniae recen, 10-20 g of dendrobium, 30-60 g of abelmoschus manihot, 10-20 g of cortex moutan, 15-30 g of salvia miltiorrhiza, 3-6 g of pseudo-ginseng powder and 10-15 g of buddleja officinalis. More preferably: 15g of astragalus, 15g of radix rehmanniae recen, 10g of dendrobium, 30g of flower of abelmoschus manihot, 15g of cortex moutan, 15g of salvia miltiorrhiza, 3g of pseudo-ginseng powder and 10g of buddleja officinalis.

Specific examples of dosage compositions are as follows:

example 1, 15g of astragalus membranaceus, 15g of radix rehmanniae recen, 10g of dendrobe, 30g of flower of abelmoschus manihot, 15g of cortex moutan, 15g of salvia miltiorrhiza, 3g of pseudo-ginseng powder and 10g of pale butterflybush flower.

Example 2, 30g of astragalus membranaceus, 10g of radix rehmanniae recen, 20g of dendrobe, 60g of flower of abelmoschus manihot, 10g of cortex moutan, 30g of salvia miltiorrhiza, 6g of pseudo-ginseng powder and 15g of butterflybush flower.

Example 3, 21g of radix astragali, 20g of radix rehmanniae, 17g of herba Dendrobii, 44g of flos Abelmoschi Manihot, 20g of cortex moutan, 19g of radix Salviae Miltiorrhizae, 5g of radix Notoginseng powder, and 13g of flos Buddlejae.

The Chinese medicinal compound preparation can be Chinese medicinal compound decoction, and the specific preparation method comprises the following steps:

secondly, adding a predetermined amount of water into the medicine decocting machine, and decocting for 30-60 minutes; squeezing out the decoction by using a mechanical squeezing part of the decoction machine; the predetermined amount is 500 and 600 ml of water corresponding to each dose of the Chinese medicinal compound preparation;

The Chinese medicinal compound preparation can also be a Chinese medicinal compound extract, and the specific preparation method comprises the following steps:

secondly, concentrating the filtrate at 65-85 ℃ under reduced pressure for 3-4 hours to obtain extract; continuously stirring the extract at 80 + -5 deg.C for 30-40 min, adding Notoginseng radix powder, and stirring for 5-10 min;

and thirdly, spray drying the obtained mixture to obtain a finished product.

The Chinese herbal medicine compound preparation can be used for preparing the medicine for treating diabetic retinopathy. The medicine has the treatment effects of inhibiting the over-expression of TGF-beta 1 protein and mRNA, VEGF protein and mRNA in retinal tissues under diabetic retinopathy and promoting the increase of miR-200b level in retinal tissues under diabetic retinopathy.

The invention is described in further detail below with reference to embodiments and with reference to the drawings. The invention is not limited to the examples given.

Examples

This example shows the pharmacodynamic data of the compound Chinese medicinal preparation of the present invention.

(1) Experimental materials:

animals: SPF grade healthy male SD (Sprague Dawley) rats, 6 weeks old, body mass around 180 g.

② a Chinese medicinal compound preparation: the prescription dose composition is as follows: 15g of astragalus membranaceus, 15g of radix rehmanniae recen, 10g of dendrobium nobile, 30g of flower of abelmoschus manihot, 15g of cortex moutan, 15g of salvia miltiorrhiza, 3g of pseudo-ginseng powder and 10g of buddleja officinalis (namely example 1). The compound traditional Chinese medicine extract (dry granular powder) is prepared by the method for standby.

(2) The experimental method comprises the following steps:

of 40 male SD rats, 8 were randomly selected as normal control groups, and 32 other rats were used to prepare DR model. The DR rats were randomly assigned to the following 4 groups after successful modeling: DR group, DR + Chinese medicinal compound low dose (FFE-L) group, DR + Chinese medicinal compound high dose (FFE-H) group, and DR + Calcium Dobesilate (CD) group.

In the DR + FFE-L group and the DR + FFE-H group, powder of the compound traditional Chinese medicine extract is prepared by 0.5 percent sodium carboxymethylcellulose solution according to the dosage of 1.0g/kg/d and 3.0g/kg/d and then is perfused into the stomach of a rat. In the DR + CD group, calcium dobesilate is prepared with 135mg/kg/d dosage and 0.5% sodium carboxymethyl cellulose solution and then is perfused into rats. The normal control group and the DR group were administered the same amount of 0.5% sodium carboxymethylcellulose solution daily for intragastric administration.

All groups were fed normally, water was taken, and the gavage was continued for 8 weeks, then all rats were sacrificed with 3% sodium pentobarbital overanesthesia, the bilateral eyeballs were rapidly removed, the cornea was cut off along the corneal scleral margin with an ophthalmic scissors, the anterior segment and the vitreous body were removed, and a cup was made. Bilateral cups of 3 mice were drawn randomly from each group, one side was stained for HE and the other side was plated for retinal vessel digestion. The rest of the cups are separated from retinal tissues, and immunofluorescence, PCR and Wb detection of TGF-beta 1 and VEGF and PCR detection of miR-200b are performed respectively.

(3) The experimental results are as follows:

influence of Chinese medicine compound preparation on pathologic change of retina tissue of DR rat

As shown in fig. 1, HE staining results showed: the control group rat retina structure is normal, the cell of the nodal cell layer is relatively regular, and the cells of the inner core and the outer nuclear layer are arranged tightly; the DR group rat ganglion cell layer cells are relatively disorderly arranged, and the inner nuclear layer is sparser and not closely arranged compared with the outer nuclear layer than the blank group. Compared with the DR group, the node cell disorder of the DR + FFE-L group is reduced, and the pathological changes of the DR + FFE-H group and the DR + CD group are obviously further improved.

② influence of Chinese medicinal compound preparation on change of retina capillary vessel of DR rat

As shown in fig. 2 and table 1, the vascularised plaques showed: the control group rat retina capillaries are regularly distributed, the diameters of the blood vessels are uniform and consistent, the trend is straight, few acellular capillaries can be seen, and the peripheral cell ghost is not seen; microvascular density (14.83 + -2.04)/HPD, E/P ratio (1.03 + -0.12).

The retina capillary network of the DR group of rats is obviously dense, disordered in walking, uneven in pipe diameter, distorted and expanded, irregular in pipe wall edge, multiple capillaries are distorted and gathered into a cluster, more acellular capillaries and pericyte ghost can be seen, and endothelial cells are obviously proliferated; both microvascular density (30.33 + -3.83)/HPD and E/P ratio (2.43 + -0.22) were significantly increased (P <0.001) compared to the control.

The microvascular changes in the DR + FFE-L group were reduced compared to the DR group, and both microvascular density (24.50 + -2.88)/HPD and E/P ratio (1.83 + -0.13) were significantly reduced compared to the DR group (P <0.01, P < 0.001).

The microvascular changes of the DR + FFE-H group and the DR + CD group are further improved, and the microvascular density and the E/P ratio are both remarkably reduced compared with the DR group (P is less than 0.001).

TABLE 1 comparison of microvascular density and E/P ratio for each group of retinal vascular digestive patches

^*vs control, P<0.01；^**vs control, P<0.001；^#vs DR，P<0.01；^##vs DR，P<0.001

③ Effect of the Chinese herbal medicine compound preparation on the expression of TGF-beta 1 and VEGF protein in DR rat retina tissue

As shown in fig. 3, retinal tissue immunofluorescence revealed: the expression of TGF-beta 1 and VEGF in DR group is obviously enhanced compared with that of control group, the expression of DR + FFE-L group is reduced compared with that of DR group, the expression of DR + FFE-H group is obviously reduced compared with that of DR group, and the expression of DR + CD group is most obviously reduced.

As shown in fig. 4, Western blot detection of retinal tissue shows that: the expression of TGF-beta 1 and VEGF protein in DR group is obviously increased (P <0.01), the expression of DR + FFE-L group is obviously reduced compared with DR group (P <0.05), the expression of VEGF in DR + FFE-H group is obviously reduced compared with DR + FFE-L group (P <0.05), and the expression of TGF-beta 1 and VEGF in DR + CD group is obviously reduced compared with DR group (P < 0.01).

Influence of Chinese medicine compound preparation on mRNA expression of miR-200b, TGF-beta 1 and VEGF in retina tissue of DR rat

As shown in fig. 5, the retinal tissue qRT-PCR assay showed: the expression of TGF-beta 1 and VEGF mRNA in the DR group is obviously increased compared with that in the control group (P <0.01), and the expression of miR-200b is obviously reduced compared with that in the control group (P < 0.01).

TGF-beta 1 and VEGF mRNA expression in DR + FFE-L group is obviously reduced compared with that in DR group (P <0.01), and miR-200b is obviously increased compared with that in DR group (P < 0.05).

The expression of VEGF mRNA in the DR + FFE-H group is further reduced compared with that in the DR + FFE-L group (P <0.01), and the expression of miR-200b is further increased compared with that in the DR + FFE-L group (P < 0.01).

The expression of TGF-beta 1 and VEGF mRNA in the DR + CD group is obviously reduced compared with that in the DR group (P is less than 0.01), and the expression of miR-200b is obviously improved compared with that in the DR group (P is less than 0.01).

It should be noted that the foregoing examples 2 and 3 also have actions or effects substantially in accordance with the aspects of example 1 described above.

The experimental result proves the effectiveness of the traditional Chinese medicine compound preparation on DR through an in-vivo experiment intervening in a DR model. The polypeptide has obvious effect of relieving the injury of retina tissues and capillaries (capillaries) of DR rats, can inhibit the over-expression of TGF-beta 1 protein and mRNA, VEGF protein and mRNA in the retina tissues of DR rats, and promotes the improvement of miR-200b level. Therefore, the traditional Chinese medicine compound has a good application prospect in DR prevention and treatment.

In terms of molecular mechanism functions:

(1) TGF-beta 1/miR-200b/VEGF pathway activation is an important molecular mechanism for DR development.

The high sugar can obviously increase the level and activity of TGF-beta 1 expressed by retinal endothelial cells, epithelial cells and the like, and the TGF-beta 1 can inhibit the activity of a miR-200b promoter to reduce the expression level of miR-200 b. miR-200b can specifically bind to VEGF mRNA, thereby inhibiting VEGF protein synthesis. High expression of VEGF in retinal tissue can specifically damage BRB (blood-retinal barrier) leading to DR development. Therefore, the high sugar enables TGF-beta 1 in the retina to be highly expressed to reduce miR-200b level, so that VEGF level is increased to damage BRB, and the pathway is regarded as an important molecular mechanism for DR generation.

(2) The traditional Chinese medicine compound preparation can obviously inhibit activation of TGF-beta 1/miR-200b/VEGF pathway.

From the foregoing, it can be seen that: the traditional Chinese medicine compound preparation can obviously inhibit the expression of mRNA and protein of TGF-beta 1 and VEGF in the retina tissue of a DR rat, and can obviously improve the expression of miR-200b, so that the traditional Chinese medicine compound preparation has obvious inhibition effect on the activation of TGF-beta 1/miR-200b/VEGF channel in the retina tissue of the DR rat. Therefore, the traditional Chinese medicine compound preparation plays a role in treating DR just because of the function of the mechanism.

Calcium dobesilate was used as a positive control in the above examples. Compared with calcium dobesilate, the traditional Chinese medicine compound preparation has the advantages that the curative effect of treating DR is equivalent, and the following advantages are achieved:

(1) good compliance

Traditional Chinese medicine has a long history, good experience and numerous patient groups in China, and even has a high degree of popularity internationally. Many patients trust and persist in using medical treatments and therefore have good compliance with medication.

(2) Various dosage forms

The Chinese medicinal compound preparation can be prepared into various dosage forms such as decoction, granules, paste, pills and the like, and can meet different requirements of different patients.

(3) Unique mechanism of action

The traditional Chinese medicine compound preparation plays a role in treating DR by inhibiting TGF-beta 1/miR-200b/VEGF pathway. The action mechanism of calcium dobesilate has been well established, and the DR treatment effect is not mainly achieved through the mechanism.

(4) Sanitary and economic advantages

The Chinese herbal medicine has excellent effect and low price, so that the Chinese herbal medicine has better sanitary economic advantages when the Chinese herbal medicine is used for treating the large medical expense of DR prevention and treatment.

In addition to the above embodiments, the present invention may have other embodiments. All technical solutions formed by adopting equivalent substitutions or equivalent transformations fall within the protection scope of the claims of the present invention.

Claims

1. A traditional Chinese medicine compound preparation for treating diabetic retinopathy is characterized by being prepared from the following traditional Chinese medicines: radix astragali, radix rehmanniae, herba Dendrobii, flos Abelmoschi Manihot, cortex moutan, radix Salviae Miltiorrhizae, radix Notoginseng powder, and flos Buddlejae; the weight ratio of the astragalus root, the dried rehmannia root, the dendrobium, the flower of abelmoschus manihot, the cortex moutan, the salvia miltiorrhiza, the pseudo-ginseng powder and the pale butterflybush flower is 15-30: 10-20: 10-20: 30-60: 10-20: 15-30: 3-6: 10-15.

2. The compound traditional Chinese medicine preparation for treating diabetic retinopathy according to claim 1, wherein the weight ratio of the astragalus root, the dried rehmannia root, the dendrobium, the flower of abelmoschus manihot, the root bark of the peony tree, the root of red-rooted salvia, the pseudo-ginseng powder and the butterflybush flower is 15: 15: 10: 30: 15: 15: 3: 10.

3. the compound traditional Chinese medicine preparation for treating diabetic retinopathy according to claim 1, which is characterized by being prepared from the following traditional Chinese medicines: 15-30 g of astragalus, 10-20 g of radix rehmanniae recen, 10-20 g of dendrobium, 30-60 g of abelmoschus manihot, 10-20 g of cortex moutan, 15-30 g of salvia miltiorrhiza, 3-6 g of pseudo-ginseng powder and 10-15 g of buddleja officinalis.

4. The compound traditional Chinese medicine preparation for treating diabetic retinopathy according to claim 3, which is characterized by being prepared from the following traditional Chinese medicines: 15g of astragalus, 15g of radix rehmanniae recen, 10g of dendrobium, 30g of flower of abelmoschus manihot, 15g of cortex moutan, 15g of salvia miltiorrhiza, 3g of pseudo-ginseng powder and 10g of buddleja officinalis.

5. The method for preparing a Chinese herbal compound preparation for treating diabetic retinopathy according to any one of claims 1 to 4, which is characterized in that the Chinese herbal compound preparation is a Chinese herbal compound decoction; the preparation method comprises the following steps:

6. The method as claimed in claim 5, wherein in the second step, the predetermined amount is 600 ml of water corresponding to each dose of the Chinese herbal compound preparation.

7. The method for preparing a Chinese herbal compound preparation for treating diabetic retinopathy according to any one of claims 1 to 4, characterized in that the Chinese herbal compound preparation is a Chinese herbal compound extract; the preparation method comprises the following steps:

and thirdly, spray drying the obtained mixture to obtain a finished product.

8. The preparation method according to claim 7, wherein the stirring time after the addition of the notoginseng powder in the second step is 5 to 10 minutes.

9. Use of the compound Chinese medicinal preparation of any one of claims 1 to 4 for preparing a medicament for treating diabetic retinopathy.

10. The use of claim 7, wherein the therapeutic effect of the medicament comprises ameliorating pathological changes in the retina under diabetic retinopathy, ameliorating retinal capillary conditions under diabetic retinopathy, inhibiting the over-expression of TGF- β 1 protein and mRNA, VEGF protein and mRNA, and promoting elevated miR-200b levels in retinal tissue under diabetic retinopathy.