Use of traditional Chinese medicine composition in preparation of medicine for preventing or treating diabetic microangiopathy

Abstract

The application relates to the technical field of traditional Chinese medicines, and discloses application of a traditional Chinese medicine composition in preparation of a medicine for preventing or treating diabetic microangiopathy, wherein the traditional Chinese medicine composition can improve blood perfusion and blood oxygen content of the foot skin of a diseased rat, reduce whole blood viscosity, has a certain protective effect on eyeball retina injury of a diabetic vasculopathy rat, inhibits infiltration of inflammatory cells in kidney tissue of a diabetic microangiopathy rat, improves glomerular basement membrane morphology, improves edema of proximal tubular epithelial cells, has a certain protective effect on foot skin injury of a diabetic microangiopathy rat, and reduces ET-1 level and Cys-C level in serum.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of traditional Chinese medicine, and particularly relates to application of a traditional Chinese medicine composition in preparation of a medicine for preventing or treating diabetic microangiopathy. BACKGROUND

[0002] Diabetic microangiopathy (DMA) is the most common vascular complication of diabetes, and its specific pathological damage mainly occurs in glomerulus, retina and peripheral microvessels. The clinical complications mainly include Diabetic Kidney Disease (DKD), Diabetic Retinopathy (DR) and Diabetic Peripheral Neuropathy (DPN), which are the main causes of blindness, kidney failure and amputation of patients.

[0003] Research shows that the DR prevalence rate increases significantly from 19.4% (course <5 years) to 78.0% (course >15 years), and is positively correlated with the course. The DKD prevalence rate of Chinese T2DM (type 2 diabetes) patients reaches 14.4%, and the DR prevalence rate is 16.5%. The screening of DPN in China shows that the DPN prevalence rate is 52.97%. DMA has become a serious problem faced by the medical field, which seriously reduces the quality of life of patients and brings huge economic burden to the families and society of patients.

[0004] At present, the drug treatment of DM (diabetes) mainly aims at comprehensive control of blood sugar, blood pressure and blood lipids, etc., but the treatment means for complications are relatively scarce.

[0005] In recent years, traditional Chinese medicine has accumulated rich clinical experience in the prevention and treatment of diabetic microangiopathy. Under the guidance of traditional Chinese medicine theory, research work has been gradually carried out in the whole country, taking the prevention and treatment of DKD, DR and DPN as the core. SUMMARY

[0006] Therefore, the present application provides application of a traditional Chinese medicine composition in preparation of a medicine for preventing or treating diabetic microangiopathy, so as to solve the problem of relatively scarce prevention and treatment means for diabetic microangiopathy.

[0007] In a first aspect, the present application provides application of a traditional Chinese medicine composition in preparation of a medicine for preventing or treating diabetic microangiopathy.

[0008] In an alternative embodiment, the traditional Chinese medicine composition comprises the following ingredients by weight: Angelica 42-48 parts, Millettia 71-79 parts, Polygoni Multiflori 145-155 parts, Red Peony Root 71-79 parts, Dipsacus 71-79 parts, Taxillus 71-79 parts, Radix Triclis 42-48 parts, Pinus Massoniana 42-48 parts, Fructus Psoraleae 57-62 parts, Radix Pseudostellariae 71-79 parts, Rhizoma Atractylodis Macrocephalae 57-62 parts, Pericarpium Citri Reticulatae 42-48 parts, Radix Aucklandiae 42-48 parts, and Red Bean 71-79 parts.

[0009] In an alternative embodiment, the diabetic microangiopathy comprises at least one of diabetic nephropathy, diabetic retinopathy, and diabetic foot skin histopathology.

[0010] In an alternative embodiment, the traditional Chinese medicine composition is Yangxuelongjin Pill.

[0011] Preferably, the traditional Chinese medicine composition is Yangxuelongjin Pill of Tongren Tang.

[0012] In an alternative embodiment, the diabetic microangiopathy is a pathological change caused by diabetes, and the diabetes is type 2 diabetes.

[0013] In an alternative embodiment, the symptoms of diabetic retinopathy include at least one of retinal capillary blood vessel distribution disorder, blood vessel basement membrane thickening, pericyte loss, and pericyte swelling.

[0014] The symptoms of diabetic nephropathy include glomerular basement membrane-like substance increase, basement membrane thickening, proximal tubular epithelial cell edema, and inflammatory cell infiltration.

[0015] The symptoms of diabetic foot skin histopathology include at least one of cell swelling and blurring, cell gap increase, and epidermal squamous epithelial cell moderate degeneration.

[0016] In an alternative embodiment, the drug for preventing or treating diabetic microangiopathy has at least one of the following functions (1)-(7):

[0017] (1) improving the pathological state of diabetic microangiopathy organ retinal blood vessels;

[0018] (2) improving the pathological state of diabetic microangiopathy organ kidney basement membrane;

[0019] (3) improving the pathological state of diabetic microangiopathy foot skin cell degeneration;

[0020] (4) reducing blood flow viscosity;

[0021] (5) improving blood oxygen saturation;

[0022] (6) improving blood flow reperfusion amount of skin at the end of the limb;

[0023] (7) reducing serum endothelin (ET-1) and cystatin (Cys-c) levels.

[0024] In an alternative embodiment, the application subject includes humans and other animals other than humans.

[0025] In an alternative embodiment, the traditional Chinese medicine composition includes Yangxuerongjin Pill or a medicine composition extracted according to the raw materials of Yangxuerongjin Pill;

[0026] Preferably, the traditional Chinese medicine composition is Yangxuerongjin Pill of Tongren Tang.

[0027] In an alternative embodiment, the medicine for preventing or treating diabetic microangiopathy further includes a pharmaceutically acceptable adjuvant.

[0028] The dosage form of the medicine for preventing or treating diabetic microangiopathy includes any preparation acceptable in pharmacy;

[0029] Alternatively, the dosage form of the medicine for preventing or treating diabetic microangiopathy is a tablet, a capsule, an oral liquid, a granule, a pill or an injection.

[0030] Compared with the prior art, the application has the following beneficial effects:

[0031] The application provides an application in the preparation of a medicine for preventing or treating diabetic microangiopathy, develops a new use of Yangxuerongjin Pill, and proves by experiments that Yangxuerongjin Pill can improve blood flow perfusion amount of skin of the foot of a diseased rat, blood oxygen content, reduce whole blood viscosity, has a certain protective effect on eyeball retina damage of a diabetic vasculopathy rat, inhibits infiltration of inflammatory cells in kidney tissue of a diabetic microangiopathy rat, improves glomerular basement membrane morphology, improves edema of proximal tubular epithelial cells, has a certain protective effect on foot skin damage of a diabetic microangiopathy rat, and reduces ET-1 level in serum and serum Cys-C level.

[0032] The above test results suggest that Yangxuerongjin Pill has a preventive and therapeutic effect on diabetic microangiopathy. BRIEF DESCRIPTION OF DRAWINGS

[0033] To more clearly illustrate the technical solutions in the specific embodiments of this application or the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this application. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0034] Figure 1 The effect of Yangxue Rongjin Pill on foot blood perfusion in model rats;

[0035] Figure 2 HE staining results of rat retinal tissue in each group;

[0036] Figure 3 PAS staining results of capillary networks in rat retinal retinal patch samples from each group;

[0037] Figure 4 HE staining results of rat kidney tissue in each group;

[0038] Figure 5 HE staining results of rat foot skin tissue in each group;

[0039] Figures 1-5 In the table, A represents the control group; B represents the model group; C represents the high-dose group of Yangxue Rongjin Pills; D represents the medium-dose group of Yangxue Rongjin Pills; and E represents the low-dose group of Yangxue Rongjin Pills. Detailed Implementation

[0040] The following embodiments are provided to better understand this application and are not limited to the preferred embodiments described herein. They do not constitute a limitation on the content and scope of protection of this application. Any product that is the same as or similar to this application, derived by anyone under the guidance of this application or by combining features of this application with other prior art, falls within the scope of protection of this application.

[0041] Unless otherwise stated or in case of contradiction, the terms or phrases used herein shall have the following meanings:

[0042] In this application, the selection range of "and / or", "or / and", and "and / or" includes any one of two or more related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that the technical solution undoubtedly includes technical solutions connected by "logical AND", and also undoubtedly includes technical solutions connected by "logical OR".

[0043] In the present application, the terms "preferably", "more preferably", "even more preferably", "advantageously" are used to describe optional features of the application and do not limit the scope of the application.

[0044] In the present application, the terms "further", "even further", "in particular" are used to describe the purpose and are not construed as limiting the scope of the application.

[0045] In the present application, the terms "optionally", "optional", "may" mean that the feature is optional, i.e. selected from either of the two parallel schemes "yes" or "no". If there are multiple "optional" in a technical solution, and there is no contradictory or mutual restrictive relationship, each "optional" is independent.

[0046] In the present application, the technical features described in an open manner include both the closed technical solution consisting of the listed features and the open technical solution containing the listed features.

[0047] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0048] In order to solve the problems existing in the prevention and treatment technology of diabetic microangiopathy, according to the first aspect of the present application, the present application provides an application of a traditional Chinese medicine composition in preparing a drug for preventing or treating diabetic microangiopathy, wherein the raw materials of the traditional Chinese medicine composition include angelica, caesalpinia, radix falcaliae, radix polygoni multiflori, radix paeoniae rubra, radix dipsaci, taxillus, radix gentianae, rhizoma smilacis glabrae, turpentine of pinus massoniana, fructus psoraleae, radix codonopsis, rhizoma atractylodis macrocephalae, pericarpium citri reticulatae, radix saussureae lappae and vigna umbellata.

[0049] In some embodiments, the diabetic microangiopathy includes at least one of diabetic nephropathy, diabetic retinopathy and diabetic foot skin histopathological changes.

[0050] In some embodiments, the traditional Chinese medicine composition is Yangxuerongjin Pill, and the traditional Chinese medicine composition is preferably Yangxuerongjin Pill of Tongren Tang.

[0051] The prescription composition of the Yangxue Rongjin Pill: Angelica 42-48 parts, Millettia nitida 71-79 parts, Radix Polygoni Multiflori (black soybean wine) 145-155 parts, Red Peony Root 71-79 parts, Dipsacus 71-79 parts, Taxillus 71-79, Radix Triclis 71-79 parts, Pinus massoniana 42-48 parts, Fructus Psoraleae (salt fried) 57-62 parts, Radix Codonopsis 71-79 parts, Atractylodes (fermented bran) 57-62 parts, Pericarpium Citri Reticulatae 42-48 parts, Radix Aucklandiae 42-48 parts, Radix Phaseoli 71-79 parts, and the auxiliary material is honey.

[0052] Function and indication: nourishing blood and invigorating sinews, dispelling wind and dredging collaterals. It is used for treating chronic diseases such as muscle and bone pain and limb numbness caused by injury.

[0053] In the prior art, it is not clear whether the Yangxue Rongjin Pill has a preventive and therapeutic effect on diseases related to diabetic microangiopathy. The present application proves through experiments that the Yangxue Rongjin Pill has the following effects:

[0054] (1) increasing the blood perfusion of the rat foot skin;

[0055] (2) increasing the oxygen content of the rat blood;

[0056] (3) reducing the whole blood viscosity;

[0057] (4) having a certain protective effect on the retinal damage of the diabetic microangiopathy rat eyeball, mainly in the inhibition of retinal thickening and the recovery of the arrangement state of each layer of cells, while reducing the gap between each layer of cells and the degree of edema, and can inhibit the thickening of the vascular basement membrane of the diabetic microangiopathy model rat to a certain extent, reduce the loss of pericytes, and reduce the swelling degree of the pericytes.

[0058] (5) having a certain protective effect on the kidney damage of the diabetic microangiopathy, mainly in the protection of organelles and the inhibition of inflammatory cell infiltration, foot process fusion and basement membrane thickening;

[0059] (6) having a certain protective effect on the foot skin damage of the diabetic microangiopathy rat, mainly in the inhibition of epidermal squamous epithelial cell degeneration and the maintenance of subcutaneous fat quantity, and the treatment effect of improving cell edema and swelling.

[0060] (7) reducing the serum ET-1 level and the serum Cys-C level.

[0061] Experiments have proved that the Yangxue Rongjin Pill has a preventive and therapeutic effect on diabetic microangiopathy; therefore, it can be known that the Yangxue Rongjin traditional Chinese medicine composition of the present application also has the above-mentioned effects and can be used for preparing a medicine for preventing or treating diabetic microangiopathy.

[0062] Dosage: oral administration. One to two pills each time, twice a day.

[0063] In some embodiments, the diabetic microangiopathy is a diabetic microangiopathy caused by type 2 diabetes.

[0064] In some embodiments, the symptoms of the diabetic retinopathy include at least one of the following: retinal capillary distribution disorder, thickening of blood vessel basement membrane, pericyte loss, pericyte swelling;

[0065] The symptoms of the diabetic kidney disease include at least one of the following: increased glomerular basement membrane-like material, thickening of the basement membrane, proximal tubular epithelial cell edema, and inflammatory cell infiltration;

[0066] The symptoms of the diabetic foot skin histopathology include at least one of the following: significant swelling of cells, increased intercellular space, and moderate degeneration of squamous epithelial cells in the epidermis.

[0067] In some embodiments, the prevention or treatment of diabetic microangiopathy is at least one of the following (1)-(7):

[0068] (1) improving the pathological state of the retinal blood vessels of the diabetic microangiopathy organ;

[0069] (2) improving the pathological state of the kidney basement membrane of the diabetic microangiopathy organ;

[0070] (3) improving the pathological state of the diabetic microangiopathy foot skin cell degeneration;

[0071] (4) reducing blood flow viscosity;

[0072] (5) increasing blood oxygen saturation;

[0073] (6) increasing the amount of skin blood flow reperfusion at the end of the limb;

[0074] (7) reducing the levels of serum endothelin (ET-1) and cystatin (Cys-c).

[0075] In some embodiments, the administration subject of application includes humans and other animals other than humans.

[0076] In some embodiments, the traditional Chinese medicine composition includes Yangxuerongjin Pills or a pharmaceutical composition extracted from the raw materials of Yangxuerongjin Pills;

[0077] When Yangxuerongjin Pills are the only effective component of the drug for preventing or treating diabetic microangiopathy, the minimum effective dose of Yangxuerongjin Pills is 1.8 g / kg / d.

[0078] Preferably, the traditional Chinese medicine composition is Yangxuerongjin Pills from Tong Ren Tang.

[0079] In some embodiments, the prophylactic or therapeutic drug for diabetic microangiopathy further comprises a pharmaceutically acceptable adjuvant.

[0080] The adjuvant includes purified water, sweeteners, emulsifiers, stabilizers, and / or vitamins, etc.

[0081] In some embodiments, the prophylactic or therapeutic drug for diabetic microangiopathy has a dosage form comprising any preparation acceptable in pharmacy;

[0082] Optionally, the prophylactic or therapeutic drug for diabetic microangiopathy has a dosage form of a tablet, a capsule, an oral liquid, a granule, a pill, or an injection.

[0083] The present application is further described in detail below with reference to specific examples, which should not be construed as limiting the scope of the application claimed.

[0084] When the specific experimental procedures or conditions are not specified in the examples, the procedures or conditions can be performed according to the conventional experimental procedures described in the literature in the art. When the reagents or instruments are not specified by the manufacturer, they are all conventional reagent products that can be purchased on the market.

[0085] Example 1

[0086] The blood-nourishing and tendon-strengthening pill used in this example has a prescription composition comprising angelica, caulis spatholobi, radix polygoni multiflori (black soybean wine), red peony root, rhizome curculiginis, radix dipsaci, radix rubi parvifolii, oil pine knot, psoralea fruit (salt-fried), radix codonopsis, fried atractylodes (fermented bran), pericarpium citri reticulatae, radix saussureae, small red beans, and honey as an adjuvant. It can be purchased from Tong Ren Tang.

[0087] 1. Experimental purpose

[0088] A diabetic microangiopathy model was established by feeding high-fat feed combined with tail vein injection of STZ. The efficacy of the blood-nourishing and tendon-strengthening pill on diabetic microangiopathy was observed through drug intervention, and the possible mechanism of action was explored.

[0089] 2. Experimental design

[0090] Fifty healthy male SD rats were adaptively fed (free access to food and water) for 4 days. Ten rats were randomly selected as a control group and fed with ordinary feed. The remaining 40 rats were fed with high-fat feed (45 kcal% fat calories) for 8 weeks, and then fasted for 16 h without water restriction before modeling.

[0091] After 16 hours of fasting without water restriction, a 2% STZ solution was prepared using a pH 4.5 citric acid-sodium citrate buffer, and 25 mg / kg was injected intravenously. Seven days later, the tail vein blood glucose value was detected. The rats that met the diabetes diagnostic criteria with a tail vein blood glucose value of ≥16.7 mmol / L were continuously fed with ordinary feed. The rats that did not meet the criteria were injected with the same dose of STZ every day. Seven days later, the tail vein blood glucose value of the rats was detected again. The rats that met the diabetes diagnostic criteria were continuously fed with ordinary feed.

[0092] The 40 rats with a blood glucose value of ≥16.7 mmol / L were randomly divided into a model group and three drug administration groups according to the blood glucose level, namely, a high-dose Yangxue Rongjin Pill group (7.2 g / kg / d), a medium-dose Yangxue Rongjin Pill group (3.6 g / kg / d), and a low-dose Yangxue Rongjin Pill group (1.8 g / kg / d). The rats in each drug administration group were given the corresponding test drug by gavage. The control group and the model group were given the same volume of distilled water. The rats in each group were continuously given the drug by gavage for 10 weeks. The general condition of the rats was observed every day, and the body weight of the rats in each group was measured every two weeks, and the corresponding drug dose was adjusted.

[0093] After the last administration of the drug, the perfusion volume / blood flow of the foot microcirculation of the rats in each group was detected using the RFLSI III laser speckle blood flow imaging system of the Rivoet company. After observing the blood flow perfusion of the rats in each group, the blood oxygen content of the rats was detected using the blood oxygen test module of the BIOPAC biological signal acquisition and processing system, and the blood oxygen saturation value was recorded.

[0094] After the detection was completed, the rats in each group were fasted for 12 hours without water restriction, and the body weight was measured. The rats were anesthetized by intraperitoneal injection of 10% chloral hydrate. After successful anesthesia, blood was taken from the abdominal aorta. One milliliter of the blood was anticoagulated with heparin and then subjected to blood rheology detection.

[0095] After the remaining blood sample was left to stand at room temperature for 2 hours, it was centrifuged at 3000 rpm and low temperature for 10 minutes. The serum was collected in an EP tube and stored in a -80°C refrigerator for standby use.

[0096] After the rats were bled, they were fixed on the rat table, and the bilateral eyeballs of the rats were enucleated in a clean environment. The left eyeball was fixed and subjected to HE staining observation. The right eyeball was subjected to retinal peeling and digestion, and then subjected to PAS staining observation. The left kidney of the rat was dissected and subjected to HE staining observation. The skin of the right foot sole of the rat was dissected, and the excess fat and fascia were removed. The skin was fixed in 4% paraformaldehyde for subsequent HE staining pathological observation.

[0097] (1) Observation of the blood flow perfusion of the feet of the rats

[0098] After the rats were anesthetized with isoflurane, they were placed in a prone position, and the toes of the feet were spread out to adjust the imaging position and distance to adapt to the irradiation area. The exposure time of the charge-coupled detector was set to 20 ms, the scanning speed was 15 fps, and the image resolution was 2064 x 1544. The collected images were converted into blood flow maps and tissue perfusion per unit time in the labeled area by the RFLSI Analysis v2.0.29.26606 system.

[0099] (2) Detection of blood oxygen content in rats

[0100] After observing the blood perfusion of the rats in each group, the blood oxygen content of the rats was detected using the blood oxygen test module of the BIOPAC biological signal acquisition and processing system, and the blood oxygen saturation value was recorded.

[0101] (3) Detection of blood rheology indexes in rats

[0102] After the administration was completed, the animals in each group were fasted for 12 h, and blood was taken from the abdominal aorta. After heparin anticoagulation, the blood rheology indexes (high shear whole blood viscosity, medium shear whole blood viscosity, and low shear whole blood viscosity) were measured using a fully automatic blood rheometer.

[0103] (4) Pathological observation of rat retina, kidney, and foot skin

[0104] After the eyeballs, kidneys, and foot skin were fixed in a paraformaldehyde buffer solution, they were flattened in a fume hood, dehydrated, embedded, and subjected to a series of processes to prepare tissue sections for hematoxylin-eosin (HE) staining. The histopathological changes were observed under a microscope. After the retina was digested, naturally dried, embedded in wax, and sectioned, PAS staining was performed to observe the lumen morphology of retinal capillaries and count and analyze endothelial cells.

[0105] (5) Detection of factors related to the pathogenesis of diabetic microangiopathy

[0106] The contents of endothelin-1 (ET-1) and cystatin C (Cys-C) in the serum of rats in each group were detected using the ELISA method according to the instructions of the kit.

[0107] 3. Experimental results

[0108] (1) Results of rat foot blood perfusion

[0109] Compared with the control group, the blood perfusion of the diabetic microangiopathy model rats was significantly reduced (P < 0.01). After drug intervention, the blood perfusion of the rats in the high, medium, and low dose groups of Yangxue Rongjin Pills was significantly increased (P < 0.01). The specific effects of Yangxue Rongjin Pills on the blood perfusion of model rats are shown in Table 1 and Figure 1 ,Figure 1 In particular, A: control group; B: model group; C: high-dose Yangxuelongjin Pill group; D: medium-dose Yangxuelongjin Pill group; E: low-dose Yangxuelongjin Pill group.

[0110] Table 1 Effect of Yangxuelongjin Pill on blood perfusion of feet of model rats

[0111]

[0112] Note: compared with the model group, **P<0.01

[0113] (2) Results of blood oxygen content detection of rats

[0114] Compared with the control group, the blood oxygen saturation of the diabetic microangiopathy model rats was significantly reduced (P<0.01), and after drug intervention, the blood oxygen saturation of the rats in the high-, medium- and low-dose Yangxuelongjin Pill groups was significantly increased (P<0.01), and the specific results are shown in Table 2.

[0115] Table 2 Effect of Yangxuelongjin Pill on blood oxygen content of model rats

[0116] Group Dose (g / kg) Number (only) Blood oxygen saturation (%) Control group — 10 84.10 ± 2.19 ** ]] Model group — 10 70.65±5.12 Nourishing blood and tendon high-dose group 7.2 10 76.95 ± 3.57 ** ]] Nourishing blood and tendon medium-dose group 3.6 10 77.64 ± 2.93 ** ]] Nourishing blood and tendon low-dose group 1.8 10 77.61 ± 5.13 ** ]]

[0117] Note: compared with the model group, **P<0.01

[0118] (3) Results of hemorheology detection of rats

[0119] After the administration was completed, the whole-automatic hemorheometer was used to determine the hemorheology indexes (high-cut whole blood viscosity, medium-cut whole blood viscosity, and low-cut whole blood viscosity). The results showed that, after modeling, compared with the control group, the high-cut, medium-cut and low-cut whole blood viscosity values of the diabetic microangiopathy model rats were significantly increased (P<0.01), and after administration, compared with the model group, the medium-cut (50) and high-cut (100 / 200) whole blood viscosity values of the rats in the medium- and low-dose Yangxuelongjin Pill groups were significantly decreased (P<0.01, P<0.05), and the specific results are shown in Table 3.

[0120] Table 3 Effect of Yangxuelongjin Pill on hemorheology of model rats

[0121]

[0122] Note: compared with the model group, *P<0.05; **P<0.01

[0123] (4) Histopathological observation of retinal tissue of rats

[0124] After modeling by high-fat diet combined with STZ, the retinas of the diabetic microangiopathy model rats were obviously thickened, the morphology of the ganglion cell layer and the cone photoreceptor cell layer was disordered and arranged irregularly, and the intercellular space of each layer was expanded and widened with obvious edema. The PAS staining results showed that the retinal capillary distribution of the model rats was disordered, the lumen of the blood vessels was uneven in thickness, and the blood vessel lumen was closed to form acellular capillaries. After drug intervention, Yangxue Rongjin Pills had a certain protective effect on the retinal injury of the diabetic vasculopathy rats, mainly in inhibiting the thickening of the retina, restoring the arrangement state of each layer of cells, and reducing the edema degree of each layer of cells. At the same time, it can inhibit the thickening of the vascular basement membrane of the diabetic microangiopathy model rats, reduce the loss of pericytes (P / E) value, and reduce the swelling degree of pericytes. The specific results are shown in Table 5, the HE staining of the retinal tissue of each group of rats (×400) is shown in Figure 2 , the retinal capillary network PAS staining (×400) of each group of rats is shown in Figure 3 , Figure 2 and Figure 3 , A: control group; B: model group; C: high-dose Yangxue Rongjin Pill group; D: medium-dose Yangxue Rongjin Pill group; E: low-dose Yangxue Rongjin Pill group.

[0125] Table 5 Effect of Yangxue Rongjin Pill on P / E value in the retinal tissue of model rats

[0126]

[0127] Note: compared with the model group, **P<0.01

[0128] From Figure 2 -A, the control group showed normal rat eyeball tissue, the overall retinal thickness was uniform, the retinal cells of each layer were clear and arranged in order, the ganglion cell layer was neat, the inner and outer nuclear layers were arranged closely, the cone photoreceptor cell layer was complete, and no abnormal histological changes such as edema and hemorrhage were observed.

[0129] From Figure 3 -A, the retinal tissue of the control group rats, the overall retinal sheet color was light, the retinal capillary distribution was regular, the direction was soft, and the tube diameter was uniform; endothelial cells were located in the center of capillary, with large nucleus and light staining; pericytes were located outside the capillary lumen, with small nucleus and deep staining.

[0130] From Figure 2 -B, it can be seen that the pathological changes occurred in the rat eyeball tissue of the model group, the retinal tissue of the model rats was obviously thickened, the thickness was uneven, the morphology of the ganglion cell layer was disordered, the arrangement was irregular, vacuolar changes were observed, the structure of the inner and outer nuclear layers was loose and disordered, the cone photoreceptor cell layer was disordered, and the intercellular space of each layer was expanded and widened with edema.

[0131] From Figure 3 -B, it can be seen that the model group can see the pathological changes of the eyeball retina tissue of the rats, the retina smear color of most samples in the model group is overall dark, suggesting that the basement membrane of the blood vessels is thickened. The distribution of the retinal capillary vessels of most samples is disordered, the lumen is uneven, the perivascular cells of some areas are swollen, the number is reduced, the arrangement is disordered, and the blood vessel lumen occlusion forms non-cellular capillary vessels.

[0132] From Figure 2 -C, it can be seen that the eyeball retina of the high-dose group of blood-nourishing and tendon-strengthening rats is slightly thickened, the vacuole-like change of the ganglion cell layer is not obvious, the arrangement is still regular, the structure of the inner and outer nuclear layers and the cone and rod cell layer is still regular, and the intercellular space of each layer is not widened. There is no obvious histological change of edema and hemorrhage.

[0133] From Figure 3 -C, it can be seen that the eyeball retina smear of the high-dose group of blood-nourishing and tendon-strengthening rats is overall light in color. The distribution of the retinal capillary vessels is still regular, and the lumen is uneven. Compared with the model group, the basement membrane thickening is improved, the capillary vessels are slightly softer, and the distribution is more regular.

[0134] From Figure 2 -D, it can be seen that the eyeball retina of the medium-dose group of blood-nourishing and tendon-strengthening rats is slightly improved in thickness, and the ganglion cell morphology is close to normal; the loose and disordered state of the inner and outer nuclear layers is slightly improved, and the arrangement of the cone and rod cell layer is obviously improved, with mild edema.

[0135] From Figure 3 -D, it can be seen that the eyeball retina smear of the medium-dose group of blood-nourishing and tendon-strengthening rats is overall light in color. The distribution of the retinal capillary vessels is still regular, and the lumen is uneven. Compared with the model group, the basement membrane thickening is improved, the capillary vessels are slightly softer, and the distribution is more regular.

[0136] From Figure 2 -E, it can be seen that the eyeball tissue of the low-dose group of blood-nourishing and tendon-strengthening rats is slightly improved in thickness; the ganglion cell morphology is close to normal, and the arrangement is still regular; the loose and disordered state of the inner and outer nuclear layers is slightly improved; and the arrangement of the cone and rod cell layer is obviously improved.

[0137] From Figure 3 -E, it can be seen that the eyeball smear of the low-dose group of blood-nourishing and tendon-strengthening rats is overall dark in color. The distribution of the retinal capillary vessels is still regular, and the lumen is uneven. Compared with the model group, the basement membrane thickening is slightly improved, the capillary vessels are slightly softer, and the distribution is more regular.

[0138] (5) Pathological observation of rat kidney tissue

[0139] After modeling, the whole kidney tissue of diabetic microangiopathy model rats showed diffuse renal damage, the glomerular basement membrane-like substance increased, and the basement membrane thickened; the proximal tubular epithelial cells showed obvious edema, and obvious ballooning degeneration occurred, the tubular basement membrane thickened, and obvious inflammatory cell infiltration was observed in part of the interstitium. After drug intervention, the glomerular basement membrane morphology of the YXJG groups of different doses was obviously improved, and the proximal tubular epithelial cell edema was reduced to varying degrees. There was no inflammatory cell infiltration in the drug groups, which could well inhibit inflammatory cell infiltration. The results of HE staining of the kidney tissue of rats in each group (x400) are shown in Figure 4 A: control group; B: model group; C: high-dose YXJG group; D: medium-dose YXJG group; E: low-dose YXJG group.

[0140] From Figure 4 A, the control group showed normal rat kidney tissue, and the whole glomerular basement membrane was normal without thickening, the mesangial matrix was not hyperplastic, the capillary wall was not thickened, the proximal tubular epithelial cells were normal, there was no edema, the tubular basement membrane was normal, there was no interstitial fibrosis, the whole kidney tubule was normal, and there was no atrophy or dilation of the kidney tubule; and there was no inflammatory cell infiltration.

[0141] From Figure 4 B, it can be seen that the model group showed pathological changes in rat kidney tissue, and the glomerular basement membrane-like substance of most samples in the model group increased, the basement membrane thickened, and involved the mesangial cells; the glomerular mesangial matrix was obviously hyperplastic, the capillary wall was thickened, the proximal tubular epithelial cells were edematous, and obvious ballooning degeneration was observed in the model group samples, the tubular basement membrane was thickened, and the interstitial fibrosis was not obvious, and the kidney tubule was dilated; part of the interstitium showed inflammatory cell infiltration. The whole showed diffuse renal damage.

[0142] From Figure 4 C, it can be seen that the glomerular mesangial matrix of the high-dose YXJG group of rats had no obvious hyperplasia, and the capillary wall had no obvious thickening. Compared with the model group, the proximal tubular epithelial cell edema of this group was less common, the tubular basement membrane thickening was not obvious, the interstitial fibrosis was not obvious, the whole kidney tubule was normal, and there was no atrophy or dilation of the kidney tubule; and there was no inflammatory cell infiltration.

[0143] From Figure 4 D, it can be seen that the glomerular mesangial matrix of the medium-dose YXJG group of rats had no obvious hyperplasia, and the capillary wall was slightly thickened. Compared with the model group, the proximal tubular epithelial cells of part of the samples in this group were edematous, the ballooning degeneration was not obvious, the tubular basement membrane had slight thickening, the interstitial fibrosis was not obvious, and the whole had no obvious inflammatory cell infiltration.

[0144] From Figure 4-E can be known, kidney tissue of rats in the low-dose group of Yangxue Rongjin can see glomerular mesangial matrix proliferation, no obvious proliferation, capillary wall slightly thickened, compared with the model group, the sample tubular basement membrane of this group has slight thickening, interstitial fibrosis is not obvious, no obvious inflammatory cell infiltration is found.

[0145] (6) Histopathological observation of rat plantar skin tissue

[0146] After modeling, the epidermis of the plantar skin of diabetic microangiopathy model rats was significantly thinned, the cells were swollen and blurred, the intercellular space was increased, and the epidermal squamous epithelial cells were moderately degenerated; the collagen fibers in the dermis layer were atrophic and degenerated, the subcutaneous fat was significantly reduced, the capillary vessels were significantly proliferated with partial expansion, the blood vessel wall was thickened, and edema and inflammatory cell infiltration were observed. After drug intervention, Yangxue Rongjin pills had a certain protective effect on the plantar skin injury of diabetic microangiopathy rats, mainly in the inhibition of the degree of degeneration of epidermal squamous epithelial cells and the maintenance of the amount of subcutaneous fat, and the improvement of the degree of cell swelling and blurring. The results of HE staining of the plantar skin tissue of rats in each group (x400) are shown in Figure 5 , A: control group; B: model group; C: high-dose Yangxue Rongjin pill group; D: medium-dose Yangxue Rongjin pill group; E: low-dose Yangxue Rongjin pill group.

[0147] From Figure 5 -A can be known, the control group shows normal rat plantar skin tissue, the overall skin tissue cell morphology is normal: the epidermis is normal in thickness, no obvious thinning is found, the epidermal cell level is not obviously reduced, no obvious cell swelling and blurring and cell space increase are found, and the cells are not degenerated, necrotic, eroded or ulcerated; normal connective tissue structure is found in the dermis layer, normal fat layer is found in the subcutaneous tissue, fat is not obviously reduced, and capillary vessels are not proliferated; no congestion, edema or inflammatory cell infiltration is found.

[0148] From Figure 5 -B can be known that the plantar skin of rats in the model group shows obvious pathological changes, which are characterized by obvious thinning of the epidermis, obvious reduction of the epidermal cell level, disappearance of the epithelial foot, and obvious cell swelling and blurring, and cell space increase. The epidermal squamous epithelial cells are moderately degenerated, and part of the epidermis lacks layered arrangement; the collagen fibers in the dermis layer are obviously atrophic and degenerated, the subcutaneous fat is obviously reduced or even disappears, the capillary vessels of part of the samples are obviously proliferated with partial expansion, forming a thick blood vessel wall; a small part of the samples shows edema and inflammatory cell infiltration.

[0149] From Figure 5 -C can be known that the plantar skin samples of rats in the high-dose Yangxue Rongjin pill group are normal in structure, the epidermis and epidermal cell level are slightly reduced, and the cells are not obviously swollen. The subcutaneous fat is slightly reduced, the capillary vessels are not obviously proliferated, and no obvious inflammatory cell infiltration is found.

[0150] From Figure 5 As can be seen from Table 4, the plantar skin sample structure of the rats in the Yangxue Rongjin Pill middle dose group was basically normal, the epidermis tissue was partially thinned, the epidermis cell hierarchy was slightly reduced, and the cell swelling degree was reduced. Compared with the model group, the dermal collagen fiber atrophy, degeneration, and subcutaneous fat reduction were improved, and no obvious inflammatory cell infiltration was observed.

[0151] From Figure 5 As can be seen from Table 4, the plantar skin sample structure of the rats in the Yangxue Rongjin Pill middle dose group was basically normal, the epidermis tissue was partially thinned, the epidermis cell hierarchy was slightly reduced, and the cell swelling degree was reduced. Compared with the model group, the dermal collagen fiber atrophy, degeneration, and subcutaneous fat reduction were improved, and no obvious inflammatory cell infiltration was observed.

[0152] (7) Microvascular lesion pathogenesis related factor detection

[0153] After modeling, the ET-1 level in the serum of the diabetic microvascular lesion model rats was significantly increased (P<0.01), and compared with the model group, the ET-1 level in the serum of the rats in the Yangxue Rongjin Pill high, middle, and low dose groups was significantly reduced (P<0.05, P<0.05, and P<0.01, respectively). After modeling, the Cys-C level in the serum of the model rats was significantly increased, and after drug intervention, the Cys-C level in the serum of the rats in the Yangxue Rongjin Pill high dose group was significantly reduced (P<0.01), and the specific results are shown in Table 6.

[0154] Table 6 Effect of Yangxue Rongjin Pill on the ET-1 and Cys-c levels in the serum of model rats

[0155]

[0156] Note: Compared with the model group, *P<0.05; **P<0.01

[0157] Obviously, the above examples are only examples for clearly illustrating, but not limiting the embodiments. For those skilled in the art, other different forms of changes or variations can be made on the basis of the above description. Here, all the embodiments need not and cannot be exhausted. The obvious changes or variations derived therefrom are still within the protection scope of the present application.

Claims

1. The application of a traditional Chinese medicine composition in the preparation of drugs for the prevention or treatment of diabetic microvascular complications, characterized in that, The traditional Chinese medicine composition consists of the following raw materials in parts by weight: Angelica sinensis 42-48 parts, Spatholobus suberectus 71-79 parts, Polygonum multiflorum (processed with black beans and wine) 145-155 parts, Paeonia lactiflora 71-79 parts, Dipsacus asper 71-79 parts, Taxillus chinensis 71-79 parts, Clematis chinensis (processed with wine and wine) 42-48 parts, Lycopodium clavatum 71-79 parts, Clematis armandii 42-48 parts, Pinus tabuliformis 42-48 parts, Psoralea corylifolia (processed with salt) 57-62 parts, Codonopsis pilosula 71-79 parts, Atractylodes macrocephala (processed with wheat bran) 57-62 parts, Citrus reticulata 42-48 parts, Aucklandia lappa 42-48 parts, Phaseolus angularis 71-79 parts, with honey as the excipient.

2. The application according to claim 1, characterized in that, The diabetic microvascular complications include at least one of diabetic nephropathy, diabetic retinopathy, and diabetic foot plantar skin histopathological changes.

3. The application according to claim 1 or 2, characterized in that, The diabetic microvascular complications are vascular complications caused by diabetes, and the diabetes is type 2 diabetes.

4. The application according to claim 3, characterized in that, The symptoms of diabetic retinopathy include at least one of the following: disordered retinal capillary distribution, thickening of the vascular basement membrane, pericyte loss, and pericyte swelling. The symptoms of diabetic nephropathy include: increased glomerular basement membrane-like material, thickening of the basement membrane, edema of proximal tubular epithelial cells, and infiltration of inflammatory cells; The symptoms of the diabetic foot plantar skin histopathology include at least one of the following: obvious cell swelling and blurring, increased intercellular spaces, and moderate degeneration of epidermal squamous epithelial cells.

5. The application according to claim 3, characterized in that, The prevention or treatment of diabetic microvascular complications is at least one of the following (1)-(7): (1) Improve the pathological state of retinal vessels in organs with diabetic microvascular disease; (2) Improve the pathological state of the renal basement membrane in diabetic microvascular disease organs; (3) Improves the pathological state of foot skin cell degeneration caused by diabetic microvascular complications; (4) Reduce blood viscosity; (5) Improve blood oxygen saturation; (6) Increase the reperfusion rate of blood flow to the skin of the extremities; (7) Reduce serum endothelin (ET-1) and cystatin (Cys-c) levels.

6. The application according to claim 1, 2, 4 or 5, characterized in that, The traditional Chinese medicine composition includes Yangxue Rongjin Pill or a drug composition obtained by extracting the raw materials of Yangxue Rongjin Pill; The prescription composition of the Nourishing Blood and Strengthening Tendon Pill, by weight, is as follows: Angelica sinensis 42-48 parts, Spatholobus suberectus 71-79 parts, Polygonum multiflorum (processed with black beans and wine) 145-155 parts, Paeonia lactiflora 71-79 parts, Dipsacus asper 71-79 parts, Taxillus chinensis 71-79 parts, Clematis chinensis (processed with wine and iron wire) 42-48 parts, Lycopodium clavatum 71-79 parts, Clematis armandii 42-48 parts, Pinus tabuliformis 42-48 parts, Psoralea corylifolia (processed with salt) 57-62 parts, Codonopsis pilosula 71-79 parts, Atractylodes macrocephala (processed with wheat bran) 57-62 parts, Citrus reticulata 42-48 parts, Aucklandia lappa 42-48 parts, Phaseolus angularis 71-79 parts, and honey as the excipient.

7. The application according to claim 1, 2, 4 or 5, characterized in that, The medications for the prevention or treatment of diabetic microvascular complications also include pharmaceutically acceptable excipients.

8. The application according to claim 1, 2, 4 or 5, characterized in that, The dosage form of the drug for the prevention or treatment of diabetic microvascular complications includes any pharmaceutically acceptable formulation.

9. The application according to claim 8, characterized in that, The dosage form is tablet, capsule, oral liquid, granule, pill, or injection.

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