Use of water extract of amomi fructus and / or vanillic acid in preparation of drug for preventing and / or treating preeclampsia

Abstract

Disclosed in the present invention is the use of a water extract of Amomi fructus and / or a vanillic acid in the preparation of a drug for preventing and / or treating preeclampsia. Further disclosed in the present invention is the use of the water extract of Amomi fructus and / or the vanillic acid in the preparation of a drug for preventing and / or treating hypertension caused by preeclampsia, or a drug for preventing and / or treating proteinuria caused by preeclampsia, or a drug for reversing a reduction in placental weight caused by preeclampsia, or a drug for restoring fetal intrauterine growth restrictions caused by preeclampsia, or a drug for restoring pathological damage to placental tissue or placental dysfunction caused by preeclampsia, or a drug for reversing the reduction in placental efficiency caused by preeclampsia, or a drug for repairing colonic barrier damage or increased intestinal permeability caused by preeclampsia, or a drug for enriching the intestinal tract of a patient with preeclampsia with Bifidobacterium bifidum. The present invention overcomes the limitations of current drugs for treating preeclampsia which exhibit limited single therapeutic effects and gastrointestinal adverse reactions.

Description

Use of water extract of litseae fructus and / or vanillic acid in preparation of medicine for preventing and / or treating pre-eclampsia TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to use of water extract of litseae fructus and / or vanillic acid in preparation of medicine for preventing and / or treating pre-eclampsia. BACKGROUND

[0002] Pre-eclampsia (PE) is a complex pregnancy-specific disease, and the main clinical features are new-onset hypertension (systolic blood pressure ≥ 140 mmHg and / or diastolic blood pressure ≥ 90 mmHg) and proteinuria occurring at 20 weeks of gestation, often accompanied by nervous system symptoms such as headache, convulsions, visual impairment, etc. When the condition is severe, it can lead to organ damage such as liver and kidney, and further cause multiple organ dysfunction, and even cause maternal and infant death. Compared with normal pregnant women, the life expectancy of patients with pre-eclampsia is shortened, and the risk of wind, cardiovascular disease and diabetes is significantly increased, and the risk of pre-eclampsia pregnancy infants developing premature birth, neurodevelopmental disorders, and cardiovascular and metabolic diseases is also significantly increased. Therefore, the prevention and treatment of pre-eclampsia is of great significance to promote reproductive health.

[0003] The pathogenesis of PE is complex and involves multiple pathological factors, including abnormal uteroplacental vascular structure, vascular endothelial dysfunction, excessive activation of inflammation and immune, and genetic factors. At present, the 'two-stage model' theory is recognized by the clinic as the main pathogenesis of PE. In the normal state of pregnancy, the uterine spiral arteries of pregnant women will expand to 5-10 times the normal size, developing into wider, broader, and higher flow rate arteries, which transport a large amount of blood, oxygen and nutrients to the fetus, ensuring the normal development of the fetus in the uterus. When preeclampsia occurs, the uterine spiral arteries of the mother are stenosis due to abnormal remodeling, which leads to insufficient blood and oxygen supply to the placenta, and further causes placental dysfunction, which is not conducive to the transport of blood oxygen and nutrients, resulting in fetal growth restriction. In order to make up for the insufficient blood oxygen supply of the placenta, the mother increases blood pressure by constricting blood vessels to drive more blood to the placenta, which leads to the occurrence of hypertension, which is one of the main clinical symptoms of PE. In addition, due to maternal placental dysfunction, a large amount of biologically active substances of placental origin are released into the maternal blood circulation, including anti-angiogenic factors, inflammatory cytokines, etc., which will induce endothelial dysfunction and inflammation in the body, further impairing the normal contraction and diastolic function of blood vessels, reducing blood flow to various organs, leading to multiple organ dysfunction, especially involving the kidney, damaging the glomerular filtration function, and causing the production of urinary protein, which is also one of the main clinical symptoms of PE. In addition, recent studies have shown that intestinal flora imbalance can lead to immune imbalance and damage to intestinal barrier function, promote the migration of pathogenic bacteria (such as Fusobacterium nucleatum) to the uterine cavity, ultimately cause placental inflammation, leading to placental dysplasia, and thus exacerbate the pathological progression of PE.

[0004] The current treatment strategies for preeclampsia mainly include preventive treatment and symptomatic treatment. According to the Clinical Trial Recommendations for Screening and Prevention of Preeclampsia in Early Pregnancy of Single Fetus published by the International Federation of Gynecology and Obstetrics (FIGO), pregnant women at high risk of preeclampsia can take low-dose (<300 mg / day) aspirin orally every day to reduce the occurrence of preeclampsia since 12-16 weeks of pregnancy. In addition, it is recommended in the Guidelines for Diagnosis and Treatment of Hypertensive Disorders in Pregnancy (2020) to control the increase of blood pressure by taking oral labetalol, nifedipine and other drugs, and to reduce the occurrence of convulsions and spasms by intravenous injection of magnesium sulfate. However, the existing treatment methods still have many adverse reactions. The preventive drug aspirin can cause gastrointestinal symptoms such as nausea, vomiting, and upper abdominal pain. Labetalol, a negative cardiac muscle strength drug, can promote pulmonary edema and heart failure and may cause bronchial spasm. In addition, symptoms of headache and nausea may occur within 24 hours after taking the drug. Nifedipine, as a calcium channel antagonist, can rapidly dilate blood vessels to lower blood pressure, but is accompanied by serious adverse reactions such as headache (especially within 24 hours after taking the drug), dizziness, flushing, palpitations, and ankle swelling. Sublingual administration of nifedipine is supported for rapid correction of severe hypertension in pregnant women, but can cause decreased placental perfusion, leading to acute fetal distress. In addition, the use of magnesium sulfate in patients with preeclampsia can reduce the risk of preeclampsia by 58%. Since intramuscular injection can cause bruises or abscesses, the best way to administer magnesium sulfate is intravenously. However, because intravenous administration is difficult to operate and requires professional medical skills, and the occurrence of preeclampsia is more common in low-income countries than in high-income countries, it is important to find a simple, safe and effective drug for the treatment of preeclampsia. SUMMARY

[0005] In order to solve the problems in the prior art, the application aims to provide the application of the aqueous extract of Amomum villosum Lour. and / or vanillic acid in the preparation of a drug for preventing and / or treating preeclampsia.

[0006] Amomum villosum Lour. is a dry mature fruit of the herbaceous plant Aomomum villosum Lour. of Zingiberaceae, and is one of the four major southern medicines in China, which has the effects of resolving dampness, warming the spleen to stop diarrhea, regulating qi and preventing miscarriage. Amomum villosum Lour. is composed of various chemical substances, including volatile components (borneol acetate, borneol, etc.) and non-volatile components (polysaccharides, flavonoids, etc.). The application finds that the aqueous extract of Amomum villosum Lour. and its main component vanillic acid play an important role in preventing and treating preeclampsia.

[0007] The specific technical solutions of the application are as follows:

[0008] The application provides the application of the aqueous extract of Amomum villosum Lour. and / or vanillic acid in the preparation of a drug for preventing and / or treating preeclampsia.

[0009] The application also provides the use of the aqueous extract of Litsea cubeba and / or vanillic acid in the preparation of a medicine for preventing and / or treating hypertension caused by preeclampsia.

[0010] The application also provides the use of the aqueous extract of Litsea cubeba and / or vanillic acid in the preparation of a medicine for preventing and / or treating proteinuria caused by preeclampsia.

[0011] The application also provides the use of the aqueous extract of Litsea cubeba and / or vanillic acid in the preparation of a medicine for reversing the decrease in placental weight caused by preeclampsia.

[0012] The application also provides the use of the aqueous extract of Litsea cubeba and / or vanillic acid in the preparation of a medicine for restoring fetal intrauterine growth restriction caused by preeclampsia.

[0013] The application also provides the use of the aqueous extract of Litsea cubeba and / or vanillic acid in the preparation of a medicine for restoring pathological damage of placental tissue or abnormal placental function caused by preeclampsia.

[0014] The application also provides the use of the aqueous extract of Litsea cubeba and / or vanillic acid in the preparation of a medicine for reversing the decrease in placental efficiency caused by preeclampsia.

[0015] The application also provides the use of the aqueous extract of Litsea cubeba and / or vanillic acid in the preparation of a medicine for repairing colon barrier damage or increased intestinal permeability caused by preeclampsia, or a medicine for enriching Bifidobacterium bifidum in the intestines of preeclampsia patients.

[0016] Further, the aqueous extract of Litsea cubeba is prepared by the following method:

[0017] 1) Grind and crush Litsea cubeba, soak it in distilled water to obtain soaked Litsea cubeba;

[0018] 2) Measure distilled water, boil it with strong fire, then add the soaked Litsea cubeba, simmer with gentle fire, filter to obtain filtrate A and dregs;

[0019] 3) Measure distilled water, boil it with strong fire, then add the dregs, simmer with gentle fire, filter to obtain filtrate B and dregs;

[0020] 4) Repeat step 3) to obtain filtrate C;

[0021] 5) Combine the filtrates, concentrate and freeze-dry to obtain Litsea cubeba freeze-dried powder, which is the aqueous extract of Litsea cubeba.

[0022] Further, the soaking time of the distilled water in step 1) is 1-6h;

[0023] And / or, the weight ratio of the distilled water to Litsea cubeba in steps 2) and 3) is 10:1-50:1;

[0024] And / or, the time of the slow-cooking in step 2) and step 3) is 5-30 min.

[0025] The beneficial effects of the present application are:

[0026] The present application simulates the pathological state of preeclampsia by subcutaneously injecting a solution of nitroso-L-arginine methyl ester (125 mg / kg), and from the 9th day of pregnancy, the mice are respectively given the aqueous extract of Amomum villosum (520 mg / kg) or vanillic acid (70 mg / kg) once a day by gavage until the end of pregnancy, and it is found that oral administration of the aqueous extract of Amomum villosum or the main component vanillic acid has a good therapeutic effect on preeclampsia.

[0027] Compared with prophylactic drugs such as aspirin and antihypertensive drugs such as labetalol, which have gastrointestinal adverse reactions, oral administration of the aqueous extract of Amomum villosum can significantly reduce the expression of inflammatory factors in the colon, increase the content of tight junction proteins to repair the intestinal barrier, and reduce intestinal permeability. At the same time, oral administration of the aqueous extract of Amomum villosum and vanillic acid can significantly enrich Bifidobacterium bifidum, a beneficial intestinal bacterium, and reshape the intestinal microecology, thus exhibiting a more comprehensive therapeutic effect and providing a new therapeutic drug for the prevention or treatment of preeclampsia.

[0028] Compared with antihypertensive drugs such as labetalol and antispasmodic drugs such as magnesium sulfate, which have a single therapeutic effect, oral administration of the aqueous extract of Amomum villosum and vanillic acid can effectively alleviate hypertension, proteinuria, placental weight reduction, placental function changes, fetal intrauterine growth restriction, and reduced placental efficiency caused by preeclampsia. In view of the limitation that existing drugs have gastrointestinal adverse reactions, oral administration of the aqueous extract of Amomum villosum can significantly reduce colon inflammation caused by preeclampsia, increase the content of tight junction proteins to repair the intestinal barrier, and reduce intestinal permeability. At the same time, oral administration of the aqueous extract of Amomum villosum and vanillic acid can significantly enrich Bifidobacterium bifidum, a beneficial intestinal bacterium, thus exhibiting a more comprehensive and safe therapeutic effect. BRIEF DESCRIPTION OF DRAWINGS

[0029] FIG. 1 is a comparison of the blood pressure of mice in each group of Example 1 (n = 6). Among them, a represents the comparison of systolic blood pressure of the NP group and the PE group on the corresponding date; b represents the comparison of systolic blood pressure of the NP and WOA groups on the corresponding date; c represents the comparison of systolic blood pressure of the NP group and the VA group on the corresponding date; d represents the comparison of systolic blood pressure of the PE group and the WOA group on the corresponding date; e represents the comparison of systolic blood pressure of the PE group and the VA group on the corresponding date; f represents the comparison of systolic blood pressure of the WOA group and the VA group on the corresponding date.

[0030] FIG. 2 is a comparison of the urine protein content of mice in each group of Example 2 (n = 6).

[0031] FIG. 3 is a comparison of the placental weight of mice in each group of Example 3 (n = 6).

[0032] Figure 4 is a comparison of the labyrinth area to junction area ratio of the placenta of each group of mice in Example 4 (n = 6), in which the scale bar is 1000 μm.

[0033] Figure 5 is a comparison of the fetal weight of each group of mice in Example 5 (n = 6).

[0034] Figure 6 is a comparison of the placental efficiency of each group of mice in Example 6 (n = 6).

[0035] Figure 7 is a comparison of the relative mRNA expression level of inflammatory factors in the colon of each group of mice in Example 7, the results of immunohistochemical staining and quantification of tight junction proteins, and the endotoxin content in serum (n = 6).

[0036] Figure 8 is a comparison of the relative mRNA expression level of Bifidobacterium bifidum in the feces of each group of mice in Example 8 (n = 6). DETAILED DESCRIPTION

[0037] In order to more clearly understand the present application, the present application will be further described with reference to the following examples and accompanying drawings. The examples are only for explanation and do not limit the present application in any way. In the examples, each of the raw reagent materials is commercially available, and the experimental methods not specifically mentioned are conventional methods and conventional conditions well known in the art, or are according to the conditions recommended by the instrument manufacturer.

[0038] 1. Pharmaceutical ingredients

[0039] (1) The water extract of Aomomum villosum Lour. (WOA) in the examples of the present application is prepared by the following preparation method:

[0040] 1) 50.0 g of Aomomum villosum Lour. is accurately weighed, crushed before use, and soaked in an appropriate amount of distilled water for 1 h;

[0041] 2) 500 mL of distilled water, which is 10 times the weight of Aomomum villosum Lour., is measured, and the soaked Aomomum villosum Lour. is added after boiling with a strong fire, and then decocted for 5 min with a weak fire, and filtered to obtain filtrate A;

[0042] 3) The residue is added again to 10 times the weight of Aomomum villosum Lour. of boiling distilled water, and then decocted for 5 min with a weak fire, and filtered to obtain filtrate B;

[0043] 4) Step 3) is repeated to obtain filtrate C;

[0044] 5) The three filtrates are combined, concentrated, and freeze-dried to obtain Aomomum villosum Lour. freeze-dried powder, which is dried and stored in a sealed container at room temperature.

[0045] (2) Vanillic acid (VA), chemical formula C8H8O4, molecular weight 168.147, is the highest content component in the water extract of Aomomum villosum Lour.

[0046] 2. Experimental animals

[0047] Male and female C57BL / 6 mice (8-10 weeks, 18-20 g) were purchased from SPF (Beijing, China) Biotechnology Co., Ltd. (Beijing, China). The mice were raised under standard light and dark cycles (12 h: 12 h) with free access to food and water. After one week of acclimatization, healthy male mice were mated with female mice at a ratio of 1:2 overnight. Female mice with sperm plugs were considered pregnant, and the day was defined as the gestational day (GD0). All animal studies were conducted in accordance with the approved protocols and guidelines of the Animal Ethics Committee of the Experimental Animal Center of Southern Medical University.

[0048] 3. Experimental reagents

[0049] Amomum villosum Lour. (Xiangchun sand) Shenzhen Institute for Drug Control Research

[0050] Vanillic acid Araldin Company, USA

[0051] Nitroso-L-arginine methyl ester Shanghai Haoyuan Biomedicine Technology Co., Ltd.

[0052] Mouse urine protein ELISA kit Jiangsu Enzyme Free Industry Co., Ltd.

[0053] Mouse serum soluble fms-like tyrosine kinase-1 (sFlt-1) ELISA kit Jiangsu Enzyme Free Industry Co., Ltd.

[0054] Mouse placental growth factor (PLGF) ELISA kit Jiangsu Enzyme Free Industry Co., Ltd.

[0055] Mouse vascular endothelial growth factor (VEGF) ELISA kit Jiangsu Enzyme Free Industry Co., Ltd.

[0056] Total RNA extraction kit for animal tissues Chengdu Fuxin Biotechnology Co., Ltd.

[0057] HiScript II Q RT SuperMix for qPCR Nanjing Novogene Bioinformatics Technology Co., Ltd.

[0058] GoTaq qPCR Master Mix Nanjing Novogene Bioinformatics Technology Co., Ltd.

[0059] 96-well fluorescent quantitative PCR plate Monoclonals Biotech Co., Ltd.

[0060] 4. Drug preparation

[0061] 1) N-nitro-L-arginine methyl ester (L-NAME) solution: 125 mg of L-NAME powder was weighed and dissolved in 10 mL of normal saline, and stored at -20℃ in the dark.

[0062] 2) Amomum villosum water extract solution: 260 mg of Amomum villosum water extract lyophilized powder was accurately weighed and dissolved in 5 mL of phosphate buffered saline solution, mixed thoroughly, and prepared for immediate use.

[0063] 3) Vanillic acid solution: 35 mg of vanillic acid powder was accurately weighed and dissolved in 5 mL of phosphate buffered saline solution, mixed thoroughly, and prepared for immediate use.

[0064] 5. Establishment and evaluation of preeclampsia model

[0065] 1) Establishment of preeclampsia model

[0066] From GD9, pregnant mice were subcutaneously injected with 125 mg / kg of L-NAME solution every day for 9 days.

[0067] 2) Evaluation of preeclampsia model

[0068] Blood pressure: the systolic blood pressure of model group mice > 110 mmHg was considered as successful modeling.

[0069] Proteinuria: the urine protein content of model group mice was significantly higher than 15.0 mg / dL, which was considered as successful modeling.

[0070] 6. Real-time fluorescent quantitative PCR

[0071] 1) Reverse transcription reaction of RNA

[0072] According to the method described in the reverse transcription kit, a total reaction system of 20 μL was configured. In a 200 μL PCR tube, the required RNA, 4 μL of HiScript II Q RT SuperMix for qPCR reagent, and ddH2O were added to make up to 20 μL. The PCR tube was placed in a PCR amplifier, and the following operation conditions were executed: 50℃, 15 min; 85℃, 5 s; 4℃, ∞. After the reaction was completed, centrifugation was performed, the time was marked, and the obtained cDNA was stored at -80℃ for standby use.

[0073] 2) Real-time fluorescent quantitative PCR (RT-qPCR)

[0074] All reverse transcription products were subjected to real-time fluorescent quantitative PCR, and the reaction system is shown in Table 1. The primer sequences of related genes are shown in Table 2, and the two-step PCR amplification system was used, and the method setting is shown in Table 3.

[0075] Table 1 PCR reaction system

[0076] Table 2 Primer sequence

[0077] Table 3 RT-qPCR reaction program

[0078] Data processing: Gapdh was used as the internal reference gene, and the relative expression of the target gene was calculated by 2-ΔΔCt method.

[0079] Example 1: Water extract of Litsea cubeba and vanillic acid can significantly restore hypertension caused by preeclampsia

[0080] Hypertension is the most critical clinical indicator of preeclampsia. This example investigates the effect of water extract of Litsea cubeba and vanillic acid on blood pressure in preeclampsia mice. In this example, mice were randomly divided into 4 groups: normal pregnancy group (NP, n = 6), preeclampsia model group (PE, n = 6), water extract of Litsea cubeba group (WOA, n = 6), and vanillic acid group (VA, n = 6). The WOA and VA groups of mice were given water extract of Litsea cubeba (520 mg / kg) and vanillic acid (70 mg / kg) by gavage from the 9th day of pregnancy (GD 9), and the normal pregnancy group and the preeclampsia model group of mice were given the same volume of 0.9% saline by gavage. Once a day, for 9 days.

[0081] The results are shown in Figure 1, and the results show that the blood pressure of the preeclampsia model group of mice is significantly higher than that of the normal pregnancy group, the water extract of Litsea cubeba group and the vanillic acid group after GD9, indicating that WOA and VA can significantly reduce the blood pressure of preeclampsia mice.

[0082] Example 2: Water extract of Litsea cubeba and vanillic acid can significantly restore proteinuria caused by preeclampsia

[0083] Proteinuria is one of the key clinical indicators of preeclampsia. In this example, subcutaneous injection of L-NAME solution was used to simulate the disease state of preeclampsia in natural occurrence, and the effects of WOA and VA on proteinuria in preeclampsia mice were investigated. In this example, the mice were randomly divided into 4 groups: normal pregnancy group (NP, n = 6), preeclampsia model group (PE, n = 6), WOA group (n = 6), and VA group (n = 6). The WOA and VA groups were administered WOA (520 mg / kg) and VA (70 mg / kg) by gavage from the 9th day of pregnancy (GD9), and the normal pregnancy group and the preeclampsia model group were administered the same volume of 0.9% saline by gavage. This was done once a day for 9 days.

[0084] The results are shown in Figure 2. The results show that the proteinuria level of the preeclampsia model mice was significantly higher than that of the normal pregnancy group, and that the proteinuria level of the model mice was significantly reduced after intervention with WOA or VA, indicating that WOA and VA can significantly reduce the proteinuria symptoms of preeclampsia model mice.

[0085] Example 3: WOA and VA can increase the placental weight of preeclampsia mice

[0086] The placenta is rich in blood vessels and nutrients, providing the necessary substances for fetal growth and development. In this example, the effects of WOA and VA on the placental weight of preeclampsia mice were investigated. In this example, the mice were randomly divided into 4 groups: normal pregnancy group (NP, n = 6), preeclampsia model group (PE, n = 6), WOA group (n = 6), and VA group (n = 6). The WOA and VA groups were administered WOA (520 mg / kg) and VA (70 mg / kg) by gavage from the 9th day of pregnancy (GD9), and the normal pregnancy group and the preeclampsia model group were administered the same volume of 0.9% saline by gavage. This was done once a day for 9 days.

[0087] The results are shown in Figure 3. The results show that the placental weight of the preeclampsia group was significantly lower than that of the normal pregnancy group, and that the decrease in placental weight was significantly reversed after intervention with WOA or VA, indicating that the intervention of WOA and VA is beneficial to the growth and development of the placenta in preeclampsia.

[0088] Example 4: WOA and VA can restore abnormal placental function

[0089] The placenta is mainly divided into three parts: the decidua region, the junction region and the labyrinth region. The labyrinth region of the placenta is mainly involved in the synthesis of hormones and cytokines (such as angiogenic factors, anti-angiogenic factors), maintaining the normal function of the placenta and fetal growth. The junction region of the placenta is mainly responsible for the exchange of oxygen, nutrients and metabolic waste. This example investigates the effects of water extract of Amomum villosum and vanillic acid on the function of the placenta of preeclampsia mice. In this example, mice were randomly divided into 4 groups: normal pregnancy group (NP, n = 6), preeclampsia model group (PE, n = 6), water extract of Amomum villosum group (WOA, n = 6), vanillic acid group (VA, n = 6). The WOA group and the VA group of mice were given water extract of Amomum villosum (520 mg / kg) and vanillic acid (70 mg / kg) by gavage from the 9th day of pregnancy (GD9), respectively, and the normal pregnancy group and the preeclampsia model group of mice were given the same volume of 0.9% normal saline by gavage. Once a day, for 9 days.

[0090] The results of HE staining are shown in Figure 4. The area ratio of the labyrinth region to the junction region of the placenta of the PE group of mice was significantly higher than that of the NP group of mice, suggesting that the placental structure of the PE mice was pathologically damaged. However, this phenomenon was reversed after intervention with WOA or VA, indicating that WOA and VA can repair the pathological damage of the placental tissue of PE mice and restore the abnormal function of the placenta.

[0091] Example 5: Water extract of Amomum villosum and vanillic acid can restore intrauterine growth restriction of fetuses of preeclampsia pregnant mice

[0092] Fetal growth restriction is one of the main complications of preeclampsia. This example investigates the effects of water extract of Amomum villosum and vanillic acid on fetal growth in preeclampsia mice. In this example, mice were randomly divided into 4 groups: normal pregnancy group (NP, n = 6), preeclampsia model group (PE, n = 6), water extract of Amomum villosum group (WOA, n = 6), vanillic acid group (VA, n = 6). The WOA group and the VA group of mice were given water extract of Amomum villosum (520 mg / kg) and vanillic acid (70 mg / kg) by gavage from the 9th day of pregnancy (GD9), respectively, and the normal pregnancy group and the preeclampsia model group of mice were given the same volume of 0.9% normal saline by gavage. Once a day, for 9 days.

[0093] The results are shown in Figure 5, which shows that the weight of the fetuses of the preeclampsia group of mice was significantly lower than that of the normal pregnancy group of mice, while the intervention of WOA or VA significantly reversed the decrease in fetal weight, indicating that WOA and VA can promote the growth and development of the fetuses of PE pregnant mice.

[0094] Example 6: Water extract of Amomum villosum and vanillic acid can significantly restore placental efficiency

[0095] Placental efficiency refers to the ratio of fetal weight to total placental weight. In preeclampsia, abnormal placental growth and development and intrauterine fetal growth restriction can cause a decrease in placental efficiency. This example investigates the effect of water extract of Amomum villosum and vanillic acid on placental efficiency in preeclampsia. In this example, mice were randomly divided into 4 groups: normal pregnancy group (NP, n = 6), preeclampsia model group (PE, n = 6), water extract of Amomum villosum group (WOA, n = 6), and vanillic acid group (VA, n = 6). The WOA and VA groups of mice were given water extract of Amomum villosum (520 mg / kg) and vanillic acid (70 mg / kg) by gavage from gestational day 9 (GD9), respectively. The normal pregnancy group and the preeclampsia model group of mice were given the same volume of 0.9% saline by gavage. This was done once a day for 9 days.

[0096] As shown in Figure 6, compared with the model group, the placental efficiency of PE pregnant mice after WOA or VA intervention was significantly increased, indicating that both WOA and VA can restore the decrease in placental efficiency induced by PE.

[0097] Example 7: Water extract of Amomum villosum can repair colon barrier damage caused by preeclampsia

[0098] In recent years, intestinal microecological imbalance has been considered as one of the pathogenesis of PE. Due to intestinal flora disorder, intestinal inflammation aggravation, and intestinal barrier damage, pathogenic bacteria are transferred from the intestine to the placenta through blood circulation, causing abnormal immune response and thus inducing PE. This example investigates the effect of water extract of Amomum villosum and vanillic acid on colon barrier damage in preeclampsia. In this example, mice were randomly divided into 3 groups: normal pregnancy group (NP, n = 6), preeclampsia model group (PE, n = 6), and water extract of Amomum villosum group (WOA, n = 6). The WOA group of mice was given water extract of Amomum villosum (520 mg / kg) by gavage from gestational day 9 (GD9). The normal pregnancy group and the preeclampsia model group of mice were given the same volume of 0.9% saline by gavage. This was done once a day for 9 days.

[0099] As shown in Figures 7A-C, compared with the model group, the expression of inflammatory factors (Il-6, Il-1β, Tnf-α) in the colon of preeclampsia mice was significantly reduced after WOA intervention, and the level of tight junction proteins (ZO-1, Occludin, and Claudin-1) was significantly increased. In addition, the intestine contains a large number of gram-negative bacteria, and endotoxin (LPS) is a heat-resistant lipopolysaccharide molecule on the cell wall of gram-negative pathogenic bacteria. When the intestinal barrier function is damaged, LPS enters the blood and is thus transferred to the surrounding tissues and organs, causing damage to the surrounding tissues and organs. As shown in Figure 7D, compared with the model group, the LPS in the serum of PE pregnant mice was significantly reduced after WOA intervention. In summary, WOA repaired the colon barrier damage induced by PE.

[0100] Example 8: Water extract of Amomum villosum and vanillic acid can significantly enrich Bifidobacterium bifidum

[0101] Bifidobacterium bifidum (B. bifidum) is an important intestinal beneficial bacteria. Maternal supplementation of B. bifidum during pregnancy can promote placental growth, nutrient transport and fetal growth and development, and participate in shaping the construction of offspring intestinal microbiota and the development of immune system. This example investigates the effect of water extract of Amomum villosum and vanillic acid on B. bifidum in the feces of preeclampsia mice. In this example, mice were randomly divided into 3 groups: preeclampsia model group (PE, n = 6), water extract of Amomum villosum group (WOA, n = 6), and vanillic acid group (VA, n = 6). The mice in WOA and VA groups were gavaged with water extract of Amomum villosum (520 mg / kg) and vanillic acid (70 mg / kg) from gestational day 9 (GD9), respectively, and the mice in the preeclampsia model group were gavaged with the same volume of 0.9% saline. This was done once a day for 9 days.

[0102] As shown in Figure 8, compared with the model group, the mRNA level of B. bifidum in the feces of PE pregnant mice was significantly increased after WOA or VA intervention, indicating that both WOA and VA significantly enriched B. bifidum.

[0103] Obviously, the above examples are merely examples for the sake of clarity, and are not limitations on the embodiments. For those of ordinary skill in the art, other different forms of changes or variations can also be made on the basis of the above description. Here, it is not necessary and impossible to exhaust all the embodiments. The obvious changes or variations derived therefrom are still within the protection scope of the present application.

Claims

1. Use of a water extract of Litsea cubeba and / or vanillic acid in the preparation of a drug for preventing and / or treating preeclampsia.

2. Use of a water extract of Litsea cubeba and / or vanillic acid in the preparation of a drug for preventing and / or treating hypertension caused by preeclampsia.

3. Use of a water extract of Litsea cubeba and / or vanillic acid in the preparation of a drug for preventing and / or treating proteinuria caused by preeclampsia.

4. Use of a water extract of Litsea cubeba and / or vanillic acid in the preparation of a drug for reversing the decrease in placental weight caused by preeclampsia.

5. Use of a water extract of Litsea cubeba and / or vanillic acid in the preparation of a drug for restoring fetal intrauterine growth restriction caused by preeclampsia.

6. Use of a water extract of Litsea cubeba and / or vanillic acid in the preparation of a drug for restoring pathological damage to placental tissue or abnormal placental function caused by preeclampsia.

7. Use of a water extract of Litsea cubeba and / or vanillic acid in the preparation of a drug for reversing the decrease in placental efficiency caused by preeclampsia.

8. Use of a water extract of Litsea cubeba and / or vanillic acid in the preparation of a drug for repairing colon barrier damage or increased intestinal permeability caused by preeclampsia, or a drug for enriching Bifidobacterium bifidum in the intestines of preeclampsia patients.

9. Use according to any one of claims 1 to 8, characterized in that, The water extract of Litsea cubeba is prepared by the following method: 1) Grind and crush Litsea cubeba, soak it in distilled water to obtain soaked Litsea cubeba; 2) Measure distilled water, boil it with strong fire, then add the soaked Litsea cubeba, simmer with gentle fire, filter to obtain filtrate A and dregs; 3) Measure distilled water, boil it with strong fire, then add the dregs, simmer with gentle fire, filter to obtain filtrate B and dregs; 4) Repeat step 3) to obtain filtrate C; 5) Combine the filtrates, concentrate and freeze-dry to obtain Litsea cubeba freeze-dried powder, which is the water extract of Litsea cubeba.

10. Use according to claim 9, characterized in that, The soaking time of the distilled water in step 1) is 1-6 hours; And / or, the weight ratio of the distilled water to Litsea cubeba in steps 2) and 3) is 10:1-50:1; And / or, the simmering time with gentle fire in steps 2) and 3) is 5-30 minutes.

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