Application of heracleum in preparation of medicine for treating and / or relieving neonatal infectious diseases and organ injury

Abstract

The invention discloses application of pubescin in preparation of a medicine for treating and / or relieving neonatal infectious diseases and visceral organ injury, after treatment with the pubescin, neonatal necrotizing enterocolitis and neonatal sepsis can be obviously relieved, the weight and survival rate of neonates can be increased, meanwhile, bacterial proliferation and infection in tissues can be obviously inhibited, and the neonatal infectious diseases and visceral organ injury can be effectively relieved. And the injury of multiple organs caused by infection is relieved. The invention can provide a new medicine and treatment choice for clinically treating neonatal necrotizing enterocolitis and neonatal sepsis, and has a wide application prospect in the aspect of treatment of neonatal infectious diseases.

Description

Technical Field

[0001] The present invention relates to the technical field of treating neonatal infectious diseases. Specifically, it relates to the application of heraclenol in the preparation of a drug for treating and / or alleviating neonatal infectious diseases and organ injuries. Background Art

[0002] Neonatal infection remains one of the important causes of morbidity and mortality among infants worldwide, especially among low-birth-weight infants, very low-birth-weight infants, and premature infants. Neonates face changes and challenges in antigen exposure after birth, including maternal contact, breastfeeding, hospital-specific microorganisms, artificial devices, drugs, nutritional antigens, etc. In addition, the immature development of the neonatal immune system, the imperfect immune function, and the immature organ development greatly increase the risk and probability of neonatal infectious diseases, posing a huge challenge to clinicians in neonatal care. As the immune antibodies obtained from the mother gradually decrease, and due to the weak function of their own immune system and the increased chance of contacting the external environment, the first week after birth is a high-incidence period for infection, and the digestive tract, respiratory tract, or oral-fecal route is the main transmission route.

[0003] Neonatal necrotizing enterocolitis (NEC) is a severe gastrointestinal disease that occurs in the neonatal period and is a relatively common gastrointestinal emergency in the neonatal intensive care unit. It is also one of the main causes of neonatal death. Clinically, it is mainly manifested by gastrointestinal symptoms such as abdominal distension, vomiting, and bloody stools. In severe cases, infectious shock, peritonitis, intestinal perforation, multiple organ failure, and even death may occur. Intestinal infection is a major factor leading to NEC. Since the neonatal digestive system is not yet fully developed, its digestive juice secretion ability, such as pancreatic juice and gastric juice, is weak, which allows a large number of bacteria to multiply in the gastrointestinal tract. In addition, the peristalsis of neonates is slow, which prolongs the colonization time of bacteria and aggravates bacterial infection, thus leading to an inflammatory response. The clinical prognosis of NEC is poor. In addition to having short-term and long-term effects on the health of children, it also brings a serious economic burden to families and society. Currently, the main clinical treatments for NEC are fasting, gastrointestinal decompression, and combined antibiotic treatment. However, the use of antibiotics may disrupt the balance of the neonatal intestinal microbiota and trigger allergic reactions. More importantly, long-term or abusive use of antibiotics can lead to the emergence of drug resistance in pathogenic bacteria, rendering antibiotics ineffective against certain infections and exacerbating the condition.

[0004] Neonatal sepsis (NS) is a severe neonatal infectious disease that can cause adverse consequences such as systemic septicemia, severe infection, purulent meningitis, and septic shock, threatening the life safety of neonates. After bacteria infect neonates, they continuously circulate in the blood and multiply in the neonates' bodies, thereby invading various organs throughout the body and producing toxins. In severe cases, it can be combined with multiple organ function damage and failure. Neonatal sepsis is mainly caused by bacterial infection, so the current clinical treatment methods are antibacterial treatment and supportive treatment.

[0005] Angelica pubescens is a plant of the genus Angelica. It has a pungent and bitter taste and is slightly warm in nature. It can warm and dredge the meridians, open the pores, and dispel wind-cold-damp pathogens in the body. Angelica pubescens has good medicinal properties and is mainly clinically applied in adjuvant treatments such as promoting blood circulation to remove blood stasis, dredging the meridians and activating collaterals, and relieving pain. Angelica pubescens has a good effect of dispelling wind and cold, and can effectively relieve symptoms such as wind-cold-damp arthralgia and joint pain. The rich nutritional components contained in Angelica pubescens can improve the nutritional status of the human body and enhance immunity. The main active ingredient of Angelica pubescens is Heraclenin. In addition to the above effects, studies have shown that the Heraclenin component in Angelica pubescens has an anti-inflammatory and swelling-reducing effect and can reduce the inflammatory response. In addition, there is no research report indicating that Heraclenin can be used for anti-bacterial infection, so the role of Heraclenin in improving neonatal infectious diseases remains unclear.

[0006] Currently, the main clinical treatment methods for neonatal infectious diseases are antibiotic treatments, and the application of traditional Chinese medicine components in the preparation of drugs for improving neonatal infectious diseases is still in its infancy. Therefore, it is of great significance to develop a traditional Chinese medicine preparation for improving neonatal infectious diseases and organ damage. Summary of the Invention

[0007] To overcome the above-mentioned defects and deficiencies in the prior art, the present invention provides the use of Heraclenin in the preparation of a drug for treating and / or relieving neonatal infectious diseases and organ damage.

[0008] The first object of the present invention is to provide the use of Heraclenin in the preparation of a drug for treating and / or relieving neonatal infectious diseases.

[0009] The second object of the present invention is to provide the use of Heraclenin in the preparation of a drug for treating and / or relieving neonatal organ damage.

[0010] The present invention claims the following:

[0011] The use of Heraclenin in the preparation of a drug for treating and / or relieving neonatal infectious diseases.

[0012] Preferably, the neonatal infectious disease is neonatal necrotizing enterocolitis.

[0013] Preferably, the neonatal infectious disease is neonatal sepsis.

[0014] Application of heraclenol in the preparation of a drug for treating and / or alleviating neonatal organ injury.

[0015] Preferably, the neonatal organ injury is caused by a neonatal infectious disease.

[0016] More preferably, the neonatal infectious disease is neonatal necrotizing enterocolitis.

[0017] More preferably, the neonatal infectious disease is neonatal sepsis.

[0018] Preferably, the organs are the intestine, liver, kidney, and / or lung.

[0019] Preferably, the drug is used to inhibit microbial proliferation and / or infection.

[0020] More preferably, the microorganism is a bacterium.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] The present invention discloses the application of heraclenol in the preparation of a drug for treating and / or alleviating neonatal infectious diseases and organ injury. After treatment with heraclenol, neonatal necrotizing enterocolitis and neonatal sepsis can be significantly alleviated, the neonatal body weight and survival rate can be increased, and at the same time, bacterial proliferation and infection in tissues can be significantly inhibited, and multi-organ injury caused by infection can be alleviated. The present invention can provide new drugs and treatment options for the clinical treatment of neonatal necrotizing enterocolitis and neonatal sepsis, and has broad application prospects in the treatment of neonatal infectious diseases. Description of the Drawings

[0023] Figure 1 Schematic diagram of the experimental design for a neonatal necrotizing enterocolitis mouse model; A: control group, B: experimental group.

[0024] Figure 2 Neonatal body weight changes, survival rate, and bacterial load in the control group and the experimental group; A: body weight change, B: survival rate, C: bacterial load in intestinal tissue.

[0025] Figure 3 Schematic diagram of the experimental design for a neonatal sepsis mouse model; A: control group, B: experimental group.

[0026] Figure 4 Neonatal body weight changes, spleen weight, and bacterial load in the control group and the experimental group; A: body weight change, B: spleen weight, C: bacterial load in spleen tissue.

[0027] Figure 5Pathological staining and histological scoring of the intestinal tissues of the control group and the experimental group of mice in Example 1.

[0028] Figure 6 Pathological staining and histological scoring of the intestinal, liver, kidney, and lung tissues of the control group and the experimental group of mice in Example 2. Detailed implementation manners

[0029] The present invention will be further described below in conjunction with specific embodiments, but the embodiments do not limit the present invention in any form. Unless otherwise specified, the reagents, methods, and equipment used in the present invention are conventional reagents, methods, and equipment in the technical field.

[0030] Unless otherwise specified, the reagents and materials used in the following examples are all commercially available.

[0031] Effect of imperatorin on neonatal necrotizing enterocolitis (NEC) in Example 1

[0032] I. Experimental method

[0033] Formula milk powder: Similac Advance infant formula milk powder (brand: Abbott Nutrition) and Esbilac puppy milk replacer (brand: PetAg) were mixed in a mass ratio of 2:1 to obtain formula milk powder.

[0034] Newborn mice were randomly divided into a control group and an experimental group, and the following treatments were carried out respectively:

[0035] Control group (NEC): Newborn mice at 7 days after birth were gavaged with formula milk powder, and at the same time, hypoxic stress was carried out 2 times a day (breathing 5% oxygen + 95% nitrogen, treatment time: 10 min) and cold stimulation was carried out 2 times a day (temperature 5°C, treatment time: 5 min). The modeling time was 4 days in total. The number of newborn mice was counted before each day of modeling, and the survival rate was calculated. The remaining newborn mice were weighed before each day of modeling, and the body weight was recorded, and the body weight change was calculated. On the 5th day of modeling, the mice were euthanized and grossly dissected, intestinal tissues were collected, and intestinal bacterial load detection was carried out ( Figure 1 A) in

[0036] Experimental group (NEC + Heraclenin): Newborn mice at 7 days after birth were gavaged with formula milk containing heraclenin (dissolved in 0.9% mass fraction physiological saline), and the administration dose of heraclenin was 30 mg / kg / 4 days. At the same time, hypoxic stress was performed twice a day (breathing 5% oxygen + 95% nitrogen, treatment time: 10 min) and cold stimulation was performed twice a day (temperature 5°C, treatment time: 5 min). The modeling time was 4 days in total. The number of newborn mice was counted before modeling every day, and the survival rate was calculated. The remaining newborn mice were weighed before modeling every day, the body weight was recorded, and the body weight change was calculated. On the 5th day of modeling, the mice were euthanized and grossly dissected, intestinal tissues were collected, and the intestinal bacterial load was detected ( Figure 1 B) in

[0037] II. Experimental results

[0038] As Figure 2 shown in A of

[0039] , compared with the body weight at the beginning of modeling, the body weight of the control group mice decreased by 19.14 ± 1.064% on the 5th day, and the body weight of the experimental group mice decreased by 12.85 ± 1.272%. Compared with the control group, the body weight change of the experimental group mice was smaller, and there was a significant difference between the two (p < 0.01). Figure 2 In terms of the survival rate, as shown in B of

[0040] , the survival rate of the control group mice was 40% on the 5th day, and the survival rate of the experimental group mice was 80%. Compared with the control group, the survival rate of the experimental group mice was significantly increased (p < 0.01). Figure 2 Since bacterial infection is one of the main causes of NEC onset, the bacterial load in the intestinal tissues of NEC mice was measured. The results are shown in C of

[0041] . Compared with the control group, the bacterial load in the intestinal tissues of the experimental group mice was significantly reduced (p < 0.05).

[0042] Example 2 Effect of heraclenin on neonatal sepsis (NS)

[0043] I. Experimental method

[0044] Newborn mice were randomly divided into a control group and an experimental group, and the following treatments were carried out respectively:

[0045] Control group (NS): Newborn mice 7 days after birth were gavaged with lipopolysaccharide solution, with a lipopolysaccharide dosage of 50 μg / kg, and the modeling time was 4 days. The remaining newborn mice were weighed before modeling every day, and their weights were recorded and the weight changes were calculated. On the 5th day of modeling, the mice were euthanized and grossly dissected, and the spleen weights were weighed and recorded, and the spleen bacterial load was tested ( Figure 3 A).

[0046] Experimental group (NS+Heraclenin): Newborn mice 7 days after birth were gavaged with lipopolysaccharide solution containing heraclenin (dissolved in 0.5% DMSO), the heraclenin dosage was 30 mg / kg / 4 days, the heraclenin dosage was 50 μg / kg, and the modeling time was 4 days. The remaining newborn mice were weighed and their body weights were recorded before modeling every day, and the weight changes were calculated. On the 5th day of modeling, the mice were euthanized and dissected, the spleen weights were weighed and recorded, and the spleen bacterial load was tested ( Figure 3 B).

[0047] 2. Experimental Results

[0048] like Figure 4 As shown in A, compared with the weight at the beginning of modeling, the weight of mice in the control group decreased by 16.67±1.667% on day 5, and the weight of mice in the experimental group decreased by 9.333±0.6667%. Compared with the control group, the weight change of mice in the experimental group was smaller, and there was a significant difference between the two (p<0.01).

[0049] The neonatal immune system is not yet fully mature. The spleen, as an important peripheral immune organ, plays a role in immune defense, immune regulation and blood filtration in the development of neonatal sepsis. Figure 4 As shown in B, after weighing the spleens of NS mice, it was found that the average weight of the spleens of the control group mice was 0.0201 g, and the average weight of the spleens of the experimental group mice was 0.0170 g. Compared with the control group, the spleen weight of the experimental group mice was significantly reduced (p<0.05).

[0050] Neonatal sepsis is mainly caused by bacterial infection, so the bacterial load in the spleen tissue of NS mice was measured. Figure 4 As shown in Figure C, the bacterial load in the spleen of mice in the experimental group was significantly reduced compared with that in the control group (p<0.05).

[0051] The above results indicate that Duhuosu can significantly improve the weight loss of mice caused by NS, and significantly reduce the weight and bacterial load of the spleen, thereby inhibiting bacterial proliferation and infection, thereby achieving the treatment of neonatal sepsis.

[0052] Example 3 Effect of Duhuosu on Organ Damage in Neonates

[0053] I. Experimental methods

[0054] Collect the visceral tissues of the control group and experimental group mice in Example 1 and Example 2, including intestinal, liver, kidney and lung tissues. Take a small section and fix it in 4% paraformaldehyde, and then perform H&E staining.

[0055] The intestinal histological score is 0-4 points, and the scoring criteria are as follows:

[0056] 0 points: The tissue villi and epithelium are intact, and the tissue structure is normal;

[0057] 1 point: Slight separation of epithelial cells from the lamina propria, but the villus structure is basically intact;

[0058] 2 points: Moderate separation of epithelial cells from the lamina propria, local villus shedding, and partial vacuolization of epithelial cells;

[0059] 3 points: Severe separation of epithelial cells from the lamina propria, most villi shedding, lamina propria edema, and a large number of vacuolizations of epithelial cells;

[0060] 4 points: The mucosa is completely damaged, the villus structure disappears, and the lamina propria is damaged.

[0061] Liver histological grading:

[0062] G0: There is no inflammation in the portal area and around the portal area;

[0063] G1: Inflammation is confined to the portal area and does not spread into the liver parenchyma. There is slight inflammation in the lobule but no hepatocyte necrosis;

[0064] G2: Mild piecemeal necrosis, and there are hepatocyte necrosis or acidophilic bodies in the lobule;

[0065] G3: Moderate piecemeal necrosis, and severe focal hepatocyte necrosis in the lobule;

[0066] G4: Severe piecemeal necrosis, and confluent necrosis can be seen in the lobule.

[0067] Kidney histology is divided into 6 grades according to the degree of brush border loss, renal tubular dilation, cast formation, renal tubular necrosis and neutrophil infiltration:

[0068] 0, normal;

[0069] 1, mild injury, involving 0%-10%;

[0070] 2, moderate injury, involving 11%-25%;

[0071] 3, severe injury, involving 26%-49%;

[0072] 4. Severe injury, involving 50% - 75%;

[0073] 5. Extensive injury, involving > 75%.

[0074] Histological grading of lung tissue:

[0075] 0. The alveolar wall is intact without thickening, no inflammatory infiltration, no congestion;

[0076] 1. Slight diffuse inflammatory cell infiltration in the alveolar wall, no obvious thickening of the alveolar wall;

[0077] 2. Obvious and extensive inflammatory cell infiltration, mild thickening of the alveolar wall (1 - 2 times);

[0078] 3. Severe inflammatory cell infiltration, thickening of the alveolar wall in individual areas up to 3 - 5 times;

[0079] 4. Severe inflammatory cell infiltration, obvious thickening of the alveolar wall, 25% - 50% of the lung tissue is parenchymal;

[0080] 5. Severe inflammatory cell infiltration, obvious thickening of the alveolar wall, > 50% of the lung tissue is parenchymal.

[0081] II. Experimental results

[0082] The results of pathological staining and histological scoring of the intestinal tissues of the control group and the experimental group of mice in Example 1 are as Figure 5 shown. The results show that in the intestinal tissues of NEC mice, moderate to severe separation of epithelial cells from the lamina propria occurs, most of the villi fall off, and most of the epithelial cells show vacuolization; after treatment with heraclenin, the villi and epithelium of the intestinal tissues are relatively intact, mostly restored to the normal tissue structure, the villus structure is basically intact, and the histological score is significantly reduced (p < 0.05).

[0083] The pathological staining of the intestines, livers, kidneys and lungs of the control group and the experimental group of mice in Example 2 is as Figure 6 shown. The results show that:

[0084] (1) In the intestinal tissues of NS mice, moderate separation of epithelial cells from the lamina propria occurs, local villi fall off, and some of the epithelial cells show vacuolization; after treatment with heraclenin, the villi and epithelium of the intestinal tissues are basically restored to the normal structure, the epithelial cells have no vacuolization, and the histological score is significantly reduced (p < 0.05).

[0085] (2) In the liver tissues of NS mice, mild piecemeal necrosis occurs, there are hepatocyte necrosis in the lobules, and some inflammatory cells appear; after treatment with heraclenin, there is no inflammation in the portal area and around the portal area of the liver tissue, the hepatocytes are normal in shape, and the histological score is significantly reduced (p < 0.05).

[0086] (3) The kidney tissues of NS mice showed mild to moderate damage, with inflammatory cell infiltration around the glomeruli, and the volume of some glomeruli was abnormally enlarged. After treatment with heraclenol, the kidney tissues basically returned to normal, without inflammatory cell infiltration, the volume and structure of the glomeruli were normal, and the histological score was significantly reduced (p<0.05).

[0087] (4) The lung tissues of NS mice showed obvious and extensive inflammatory cell infiltration, obvious red blood cell infiltration, and moderate to severe thickening of the alveolar wall. After treatment with heraclenol, there was basically no inflammatory cell and red blood cell infiltration in the lung tissues, the alveolar wall was not thickened, and the histological score was significantly reduced (p<0.05).

[0088] The above results indicate that heraclenol can significantly alleviate neonatal organ damage caused by neonatal necrotizing enterocolitis and neonatal sepsis.

[0089] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited by the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. The use of Duhuosu in the preparation of drugs for treating and / or alleviating neonatal infectious diseases.

2. The use according to claim 1, characterized in that: The neonatal infectious disease is neonatal necrotizing enterocolitis.

3. The use according to claim 1, characterized in that: The neonatal infectious disease is neonatal sepsis.

4. The use of Duhuosu in the preparation of drugs for treating and / or alleviating neonatal organ damage.

5. The use according to claim 4, characterized in that: The neonatal organ damage is caused by neonatal infectious diseases.

6. The use according to claim 5, characterized in that: The neonatal infectious disease is neonatal necrotizing enterocolitis.

7. The use according to claim 5, characterized in that: The neonatal infectious disease is neonatal sepsis.

8. The use according to claim 4, characterized in that: The internal organs are intestines, liver, kidneys and / or lungs.

9. The use according to any one of claims 1 to 8, characterized in that: The drug is used to inhibit microbial proliferation and / or infection.

10. The use according to claim 9, characterized in that: The microorganisms are bacteria.

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