Use of Xiaojin pills in the preparation of a medicament for treating and / or preventing abdominal aortic aneurysm

Abstract

The application relates to the technical field of biological medicines, in particular to the use of Xiaojin pills in the preparation of medicines for treating and / or preventing abdominal aortic aneurysm. The Chinese patent medicine Xiaojin pills are marketed medicines, which are administered in an oral manner. The application proves through animal experiments that Xiaojin pills can be used in the medicines for treating and / or preventing abdominal aortic aneurysm, and provides an effective new potential alternative method for the prevention and treatment of abdominal aortic aneurysm.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, and particularly relates to the use of Xiaojin Pills in the preparation of a medicine for treating and / or preventing abdominal aortic aneurysm. BACKGROUND

[0002] Abdominal aortic aneurysm (AAA) is a fatal cardiovascular disease, which is characterized by permanent expansion of the abdominal aorta, and the mortality rate after rupture is about 80%. If not treated effectively, AAA will continue to grow until the aneurysm ruptures. Age, male, AAA family history, smoking and hypertension are all risk factors associated with AAA.

[0003] However, so far, there is no drug in clinic that can effectively cure this disease. For patients with aortic diameter greater than 5.5 cm, surgical repair is recommended, and for patients with aortic diameter of 3.0-5.4 cm, it is defined as small AAA, which can only be closely monitored. Although there has been progress in surgical selection and several drugs, several drugs such as doxycycline, ACEI, statins and others have been proposed to prevent AAA, but no proven drug can be used for drug intervention of AAA.

[0004] Therefore, it is necessary to find a drug health care to prevent the occurrence of AAA and effectively inhibit the expansion of AAA. SUMMARY

[0005] The present application provides the use of Xiaojin Pills in the preparation of a medicine for treating and / or preventing abdominal aortic aneurysm.

[0006] The present application uses elastase and angiotensin II to construct two mouse abdominal aortic aneurysm models, and gives different concentrations of Xiaojin Pills drug treatment, and detects the growth inhibition effect of Xiaojin Pills on mouse abdominal aortic aneurysm. The detection results show that Xiaojin Pills can inhibit the growth and incidence of abdominal aortic aneurysm. Therefore, Xiaojin Pills can be used in the medicine for treating and / or preventing abdominal aortic aneurysm.

[0007] The medicine can inhibit the diameter of the abdominal aorta.

[0008] The medicine can inhibit the incidence of abdominal aorta.

[0009] The Xiaojin Pills have the function of protecting vascular smooth muscle cells.

[0010] The Xiaojin Pills have the function of protecting elastic fibers and inhibiting the rupture and degradation of elastic fibers.

[0011] The Xiaojin Pills have the function of inhibiting aneurysm formation.

[0012] Optionally, the abdominal aortic aneurysm is elastase-induced abdominal aortic aneurysm.

[0013] Optionally, the abdominal aortic aneurysm is angiotensin II-induced abdominal aortic aneurysm.

[0014] In summary, the beneficial technical effects of the present application are as follows:

[0015] The present application proves by animal experiments that Xiaojin Pill can be used for treating and / or preventing abdominal aortic aneurysm in drugs, and provides an effective new potential alternative method for the prevention and treatment of abdominal aortic aneurysm. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 Experimental results of each group of elastase model mice (control - blank treatment group; AAA - model group; NGN - positive drug control group, 50 mg / kg / day; XJW(H) - Xiaojin Pill high-dose group, 900 mg / kg / day; XJW(L) - Xiaojin Pill low-dose group, 300 mg / kg / day; A is the picture of the material at the end of the experiment; B is the comparative analysis of the abdominal aortic diameter, Mean ± SEM, *P<0.05, **P<0.01).

[0017] Figure 2 HE staining and EVG staining results of each group of elastase model mice.

[0018] Figure 3 Body weight changes of each group of elastase model mice on days 1, 5, 9, and 13.

[0019] Figure 4 Experimental results of each group of angiotensin II pump model mice (control - blank treatment group; Ang II - model group; Ang II + NGN - positive drug control group, 50 mg / kg / day; Ang II + XJW - Xiaojin Pill low-dose group, 300 mg / kg / day; A is the picture of the material at the end of the experiment; B is the comparative analysis of the abdominal aortic diameter, Mean ± SEM, *P<0.05, **P<0.01).

[0020] Figure 5 Statistical results of abdominal aortic aneurysm incidence between each group of angiotensin II pump model mice (Chi-square, *P<0.05).

[0021] Figure 6 Blood pressure of each group of angiotensin II pump model mice.

[0022] Figure 7 HE staining and EVG staining results of each group of angiotensin II pump model mice.

[0023] Figure 8 The weight change of each group of mice with implanted angiotensin II pump model on the 1st, 8th, 15th and 22nd day. DETAILED DESCRIPTION

[0024] Before describing the embodiments of the present application in detail, it should be understood that the terms used herein are for the purpose of describing specific embodiments only. 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 the term belongs.

[0025] Xiaojin Pill is a pill formula of Wang Hongxu, a famous doctor in Qing Dynasty, which can treat yin ulcer such as thyroid nodule, uterine fibroids, ovarian cysts, etc. The main components of Xiaojin Pill are artificial musk, Gleditsia sinensis L. (without shell and oil), prepared Aconitum, tolu balsam, frankincense (prepared), myrrh (prepared), five spirit fat (vinegar fried), angelica (wine fried), earthworm, and ink. Xiaojin Pill can disperse and remove swelling, remove blood stasis and relieve pain, and is used for treating scrofula, goiter, breast mass, breast mass caused by phlegm and qi stagnation, with one or several lumps under the skin or skin, which can move when pushed, or bone and joint swelling, skin color unchanged, and hard and painful lumps.

[0026] The present application finds that Xiaojin Pill is significantly negatively correlated with the gene expression disorder of abdominal aortic aneurysm through bioinformatics analysis, so Xiaojin Pill can possibly reverse the abdominal aortic aneurysm disorder gene, further indicating that Xiaojin Pill can possibly be used in drugs for treating and / or preventing abdominal aortic aneurysm.

[0027] Based on this, the present application further carries out a feasibility study.

[0028] The present application first studies the correlation between Xiaojin Pill and abdominal aortic aneurysm. Through the study, it is found that Xiaojin Pill can be used in drugs for treating and / or preventing abdominal aortic aneurysm.

[0029] In the present application, the drug Xiaojin Pill is prior art and can be purchased on the market.

[0030] The present application provides the use of Xiaojin Pill in preparing a drug for treating and / or preventing abdominal aortic aneurysm.

[0031] The drug can inhibit the diameter of the abdominal aorta.

[0032] The drug can inhibit the incidence of abdominal aortic aneurysm.

[0033] The abdominal aortic aneurysm is an abdominal aortic aneurysm caused by elastase.

[0034] The abdominal aortic aneurysm is an abdominal aortic aneurysm caused by angiotensin II.

[0035] The Xiaojin Pill has the function of protecting vascular smooth muscle cells.

[0036] The Xiaojin Pill has the effect of protecting elastic fibers and inhibiting the breaking and degradation of elastic fibers.

[0037] The Xiaojin Pill has the effect of inhibiting the formation of tumors.

[0038] In order to make the purpose, technical solutions and advantages of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below in combination with the drawings. Based on the embodiments of the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0039] Unless otherwise specified, the technology or conditions in the embodiments are carried out according to the technology or conditions described in the literature in the art or according to the product manual. The reagents or instruments used are not specified by the manufacturer, but are conventional products that can be obtained commercially.

[0040] The present application will be further described in detail below in combination with the embodiments, drawings and test results.

[0041] In the following embodiments, the test animals include C57BL / 6J mice, ApoE − / − mice.

[0042] C57BL / 6J mice, 20-25g, SPF level, male, purchased from the Experimental Animal Center of Peking University Health Science Center.

[0043] ApoE − / − mice, 30-35g, SPF level, male, purchased from the Experimental Animal Center of Peking University Health Science Center.

[0044] In the following embodiments, the manufacturer of Xiaojin Pill is Chengdu Yongkang Pharmaceutical Co., Ltd.; the manufacturer of the positive control drug naringenin is sigma (USA); the manufacturer of elastase is sigma (USA); and the manufacturer of angiotensin II is MCE (USA). Embodiment Embodiment 1

[0045] This embodiment explores the inhibitory effect of Xiaojin Pill on the elastase model group.

[0046] Specifically includes the following processes:

[0047] (1) Experimental grouping

[0048] The experimental animals were randomly divided into a blank treatment group (control), a model group (AAA), a positive drug control group (NGN, 50 mg / kg / day), a Xiaojinwan high-dose group (XJW(H), 900 mg / kg / day), and a Xiaojinwan low-dose group (XJW(L), 300 mg / kg / day).

[0049] (2) Medication and administration method

[0050] Naringenin: Take an appropriate amount and dissolve in physiological saline to configure 16 mg / mL. Prepare fresh and store in the dark.

[0051] Xiaojinwan: Take an appropriate amount of pills and crush them, put them in a glass bottle containing a rotor, add an appropriate amount of physiological saline, and heat and stir for 30 min to configure 0.3 g / mL. Prepare fresh and use immediately.

[0052] The administration method is as follows:

[0053] The administration method is as follows:

[0054] Model group: Each animal was given an equal amount of physiological saline by gavage every day.

[0055] Positive drug control group: Each animal was given a dose of 50 mg / kg / day by gavage.

[0056] Xiaojinwan high-dose group: Each animal was given a dose of 900 mg / kg / day by gavage.

[0057] Xiaojinwan low-dose group: Each animal was given a dose of 300 mg / kg / day by gavage.

[0058] (3) Abdominal aortic aneurysm modeling - elastase model

[0059] 1) High-pressure surgical instruments, cosmetic cotton, glove strips, sterile physiological saline were prepared before the experiment;

[0060] 2) 8-10 week old mice were anesthetized with 1% sodium pentobarbital (about 1.5 ml per mouse) by intraperitoneal injection and fixed on the mouse plate with tape;

[0061] 3) The abdomen was shaved with depilatory cream, and after the depilatory cream was applied, it was left for 3-5 min, and the mouse hair was removed with toilet paper;

[0062] 4) Open the abdomen, carefully cut open the abdominal skin, gently stir the space with a cotton swab, find the infrarenal abdominal aorta, gently tear the external connective tissue of the blood vessel, and separate the infrarenal abdominal aorta;

[0063] 5) With tweezers, pass the cut 0.5 cm wide glove strip under the blood vessel, pass the previously cut 0.3-0.5 cm wide cosmetic cotton strip under the blood vessel, place the glove strip between the blood vessel and the cosmetic cotton strip, remove the excess cosmetic cotton stained with blood, and try to control the amount of blood on the cosmetic cotton wrapped around the blood vessel;

[0064] 6) Add 30 ul of elastase (4.5 U) on top of the mouse renal inferior abdominal aorta through the glove strip and cosmetic cotton, and wrap with cosmetic cotton and glove strip for 40 min;

[0065] 7) Remove the cosmetic cotton and glove strip, flush the abdominal cavity with a small amount of clean normal saline with a syringe, and add normal saline containing penicillin on the wound to prevent infection, and suture the wound. After 14 days, take the sample, anesthetize, take blood, perfuse and fix, separate the mouse aorta, and place it in paraformaldehyde; take a general photograph.

[0066] (4) Observation index

[0067] Abdominal aortic diameter expansion to 1.5 times or more of the normal diameter is defined as abdominal aortic aneurysm.

[0068] Elastase model group: During the experiment, pay attention to the activity, diet, etc. of each mouse, weigh each mouse every 4 days, and measure the abdominal aorta diameter of each mouse after taking the sample.

[0069] (5) Statistical method

[0070] The abdominal aorta diameter of each blood vessel was measured using ImageJ software, and data statistical analysis was performed using Graphpad, One-way ANOVA, *p<0.05, **p<0.01; Chi-square was used to calculate the difference between the incidence rates of each group, *p<0.05, **p<0.01. Example 2

[0071] This example explores the inhibitory effect of Xiaojin Pill on the angiotensin II pump model.

[0072] Specifically includes the following processes:

[0073] (1) Experimental grouping

[0074] The experimental animals were randomly divided into a blank treatment group (control), a model group (Ang II), a positive drug control group (Ang II+NGN, 50 mg / kg / day), and a low-dose Xiaojin Pill group (Ang II+XJW, 300 mg / kg / day).

[0075] (2) Drug and administration method

[0076] Naringenin: Take an appropriate amount and dissolve in physiological saline to configure 16 mg / mL. Prepare fresh and store in the dark.

[0077] Xiaojin Pill: Take an appropriate amount of pill body and crush it, put it into a glass bottle with a rotor, add an appropriate amount of physiological saline, heat and stir for 30 min, and configure it into 0.3 g / mL. Prepare fresh and use.

[0078] The administration method is as follows:

[0079] The model mice were administered seven days before modeling.

[0080] Model group: Each animal was given an equal amount of physiological saline by gavage every day.

[0081] Positive drug control group: Each animal was given a dose of 50 mg / kg / day by gavage.

[0082] Xiaojin Pill low-dose group: Each animal was given a dose of 300 mg / kg / day by gavage.

[0083] (3) Abdominal aortic aneurysm modeling - angiotensin II pump implantation model

[0084] Ang II pump; the amount used for each mouse is 1000 ng / kg / min, the Ang II mother liquor is prepared according to the weight of the largest mouse, and the rest is diluted with an appropriate amount of physiological saline (dissolved in physiological saline on a clean bench, and the solution is placed on ice). The Ang II solution is injected into the slow-release pump with a syringe, and the pump cover is inserted. The pump is soaked in physiological saline for 12 hours before surgery and placed in a 37°C incubator.

[0085] Pump implantation surgery:

[0086] 1) Sterile surgical instruments, needles, sterile physiological saline, 75% alcohol, beaker, dish, 1% pentobarbital sodium solution, cotton swab, 1 mL syringe, etc. are prepared before surgery;

[0087] 2) The mouse is anesthetized with 1% pentobarbital sodium;

[0088] 3) Shave the neck and back, and disinfect the skin with alcohol;

[0089] 4) Use scissors to open the mouth, use a needle holder to expand subcutaneously, and place the slow-release pump cover inside;

[0090] 5) Suture the wound, observe the mice daily after surgery (whether there is death), and gently rub the location of the pump every day to prevent the pump from adhering to the skin;

[0091] 6) After 28 days, take the material after weighing the body weight and measuring the blood pressure, anesthetize and take blood, perfuse and fix, separate the mouse aorta and place it in paraformaldehyde; take a general photograph.

[0092] (4) Observation index

[0093] The abdominal aortic diameter expansion to 1.5 times and above the normal diameter is defined as abdominal aortic aneurysm.

[0094] Angiotensin II pump group: During the experiment, pay attention to the death of each mouse, record whether each dead mouse died of aortic aneurysm rupture, and measure the blood pressure of each mouse before sampling. Every 7 days, weigh each mouse, measure the abdominal aortic diameter of each mouse after sampling, and count the incidence and other indicators.

[0095] (5) Statistical method

[0096] The abdominal aortic diameter of each blood vessel was measured by ImageJ software, and the data was statistically analyzed by Graphpad, using One-way ANOVA, *p<0.05, **p<0.01; the difference between the incidence of each group was calculated by Chi-square, *p<0.05, **p<0.01.

[0097] Detection results

[0098] The detection results of Example 1 and Example 2 are shown in Table 1 and Figures 1-8 .

[0099] Figure 1 The experimental results of the elastase model mice in each group. Among them, Figure A is the sampling photo of the elastase model mice at the end of the experiment, and Figure B is the comparative analysis of the abdominal aortic diameter of the elastase model mice. From Figure 1 it can be seen that the abdominal aortic diameter of the positive drug control group naringin is significantly lower than that of the model group (p<0.01), and the high-dose Xiaojin Pill (p<0.01) and low-dose Xiaojin Pill group (p<0.05) can also significantly reduce the abdominal aortic expansion of the elastase-induced mice.

[0100] Figure 2 The results of HE staining and EVG staining of the elastase model mice in each group. From Figure 2It can be seen from the results of HE staining that the control group is normal aorta, and the smooth muscle cell layer is tightly and firmly connected by elastic fibers and collagen fibers, while the AAA group is irregular in shape, and the vascular media almost disappears due to the degradation of matrix proteins in the wall, resulting in decreased elasticity. The vascular media is well preserved in the XJW(H) and XJW(L) groups, and the smooth muscle cell layer does not show obvious loosening and disappearance, indicating that Xiaojin Pill has a good protective effect on the function of vascular smooth muscle cells. At the same time, from the results of EVG staining, it can be seen that the elastic fibers in the control group are deeply colored, and the aortic elastic fiber content is normal, while the elastic fibers in the AAA group are the least colored, and the aortic elastic fiber content is significantly reduced, and the reduction of elastic fibers is significantly inhibited in the XJW(H) and XJW(L) groups, indicating that Xiaojin Pill can protect elastic fibers and inhibit their degradation.

[0101] Figure 3 is the weight change of each group of elastase model mice on the 1st, 5th, 9th and 13th day. From Figure 3 It can be seen that there is no significant difference in weight change among the 5 groups of mice.

[0102] Figure 4 is the experimental results of each group of angiotensin II pump implantation model mice. Among them, A is the photograph of the material at the end of the experiment of angiotensin II pump implantation model mice, and B is the comparative analysis of the diameter of the abdominal aorta of angiotensin II pump implantation model mice. From Figure 4 It can be seen that the abdominal aorta diameter of the positive drug control group naringin is significantly lower than that of the model group (p<0.01), and the low dose group of Xiaojin Pill (p<0.05) can also significantly reduce the abdominal aorta dilation of mice caused by angiotensin II.

[0103] Figure 5 is the statistical results of the incidence of abdominal aortic aneurysm among each group of angiotensin II pump implantation model mice. From Figure 5 It can be seen that the incidence of the model group is 70%, and the incidence of the positive drug control group naringin is 26%, which can significantly reduce the incidence of abdominal aortic aneurysm caused by angiotensin II (p<0.01), and the incidence of the low dose group of Xiaojin Pill is 36%, which can also significantly reduce the incidence of abdominal aortic aneurysm caused by angiotensin II (p<0.05).

[0104] Figure 6 is the blood pressure of each group of angiotensin II pump implantation model mice. From Figure 6 It can be seen that there is no significant difference in blood pressure among the 4 groups of mice.

[0105] Figure 7 is the results of HE staining and EVG staining of each group of angiotensin II pump implantation model mice. From Figure 7It can be seen from the results of HE staining and EVG staining that the elastic fibers in the aorta of the control group are arranged closely, while the elastic fiber plates in the model group are broken and even disappear, and obvious tumor formation is observed. The low-dose Xiaojinwan group can significantly inhibit the rupture of elastic fibers and the formation of tumors.

[0106] Figure 8 The weight changes of the angiotensin II pump model mice in each group on days 1, 8, 15, and 22 are shown in the following table. Figure 8 It can be seen that there is no significant difference in the weight changes of the five groups of mice.

[0107] Table 1 Anti-abdominal aortic aneurysm effect of Xiaojinwan

[0108]

[0109] From Table 1, compared with the model group, the positive drug control group naringin can significantly reduce the diameter and incidence of abdominal aortic aneurysm, p<0.01, and has no significant effect on the blood pressure and body weight of mice. Combined with Table 1 and Figures 1-3 It can be seen that the high-dose Xiaojinwan group can significantly inhibit the diameter of abdominal aortic aneurysm in elastase model mice, p<0.01, and has no significant effect on the body weight of mice. Combined with Table 1 and Figures 5-8 It can be seen that the low-dose Xiaojinwan group can significantly inhibit the diameter and incidence of abdominal aortic aneurysm in angiotensin II pump model mice, p<0.05, and has no significant effect on the blood pressure and body weight of mice.

[0110] In summary, the Chinese medicine Xiaojinwan has an inhibitory effect on the occurrence of abdominal aortic aneurysm caused by elastase and angiotensin II.

[0111] Finally, it should be noted that the above examples are only used to illustrate the technical solutions of the present application, and are not limited thereto; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that they can still modify the technical solutions described in the foregoing examples, or make equivalent replacements to some technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. Use of Xiaojin pills in the preparation of a medicament for treating and / or preventing abdominal aortic aneurysm, characterized in that, The main components of the small gold pill are artificial musk, shelled and deoiled semen astragali, prepared eupatorium fortunei, toon resin, prepared frankincense, prepared myrrh, vinegar-fried five musk, wine-fried angelica, earthworm, and fragrant ink.

2. The use of Xiaojin pills according to claim 1 in the preparation of a medicament for treating and / or preventing abdominal aortic aneurysm, characterized in that, The medicine can inhibit the diameter of abdominal aorta.

3. The use of Xiaojin pills according to claim 1 in the preparation of a medicament for treating and / or preventing abdominal aortic aneurysm, characterized in that, The medicine can inhibit the incidence of abdominal aorta.

4. The use of Xiaojin pills according to claim 1 in the preparation of a medicament for treating and / or preventing abdominal aortic aneurysm, characterized in that, The abdominal aortic aneurysm is caused by elastase.

5. The use of Xiaojin pills according to claim 1 in the preparation of a medicament for treating and / or preventing abdominal aortic aneurysm, characterized in that, The abdominal aortic aneurysm is caused by angiotensin II.

6. The use of Xiaojin pills according to claim 1 in the preparation of a medicament for treating and / or preventing abdominal aortic aneurysm, characterized in that, The small gold pill has the function of protecting vascular smooth muscle cells.

7. The use of Xiaojin pills according to claim 1 in the preparation of a medicament for treating and / or preventing abdominal aortic aneurysm, characterized in that, The small gold pill has the function of protecting elastic fibers and inhibiting the rupture and degradation of elastic fibers.

8. The use of Xiaojin pills according to claim 1 in the preparation of a medicament for treating and / or preventing abdominal aortic aneurysm, characterized in that, The small gold pill has the function of inhibiting the formation of aneurysms.

Images