Application of crocin II in preparation of medicine for preventing or treating sepsis renal injury

Abstract

The invention belongs to the technical field of medicines, and particularly relates to application of crocin II in preparation of a medicine for preventing or treating sepsis renal injury. According to the application disclosed by the invention, experimental research is carried out on the crocin II in the aspect of preventing or treating renal injury caused by sepsis by constructing a sepsis model, and a result shows that the crocin II has a good protection effect on a sepsis multi-organ injury model induced by a cecum ligation puncture (CLP) method and is small in side effect; the compound can be used for preparing protective drugs for kidney injury caused by sepsis. The invention provides an application of crocin II in preparation of a therapeutic drug for kidney injury caused by sepsis, and the crocin II has obvious effects of improving acute kidney injury and playing an anti-inflammatory effect and is small in side effect.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and particularly relates to use of crocin II in preparing a medicine for preventing or treating sepsis-induced kidney injury. Background Art

[0002] Sepsis is a serious systemic infection caused by a variety of pathogens, including bacteria, invading the human body through various pathways, which in turn leads to a systemic inflammatory response syndrome. The mechanisms of sepsis are complex and diverse, including systemic inflammatory response, immune dysfunction, abnormal coagulation function, tissue damage, and abnormal responses of the body to pathogenic microorganisms and endotoxins, involving pathological and physiological changes in multiple systems and organs. The most common organ damage includes brain damage, lung damage, kidney damage, and liver damage.

[0003] Sepsis-associated acute kidney injury (SA-AKI) is a common complication in critically ill patients with high morbidity and mortality. SA-AKI is generally defined as sepsis or disruptive shock involving the kidneys, which leads to a progressive decline in renal function. The pathophysiology of SA-AKI involves damage and dysfunction of multiple cell types, including macrophages, vascular endothelial cells (ECs), and tubular epithelial cells (TECs), as well as the interactions and associations between them. Increasing evidence suggests that the pathogenesis of SA-AKI is multifactorial and complex, involving interactions between inflammation, microcirculatory dysfunction, and metabolic reprogramming. Moreover, sepsis and renal injury make patients mutually susceptible, and it is often difficult to determine the exact onset of these two syndromes clinically. There is currently a great need for drugs that can effectively prevent and treat septic renal injury.

[0004] Crocus is the dried stigma of the perennial herb Crocus sativus L. of the Iridaceae family, which has the effects of promoting blood circulation and removing blood stasis, cooling blood and detoxifying, relieving depression and calming the nerves. It is commonly used for amenorrhea, postpartum blood stasis, fever, rashes, depression, palpitations and madness. Crocus can be used in traditional Chinese medicine decoctions for hair growth, but there are few research reports on its effective active ingredients for promoting hair growth. Crocin, also known as crocin, crocin or safranin, is the main component of crocin. Crocin is a series of ester compounds formed by its aglycone crocetin and gentiobiose or glucose, mainly composed of crocin I, crocin II, crocin III, crocin IV and crocin V. Modern pharmacological studies have shown that crocin has good therapeutic effects on a variety of central nervous system diseases, such as neurodegenerative diseases, mental disorders, epilepsy, convulsions and insomnia, as well as a variety of cardiovascular system diseases, such as hypertension, hyperlipidemia and atherosclerosis. It also has anti-cancer, anti-inflammatory, antioxidant, liver-protective and choleretic, and anti-diabetic effects. Therefore, the medicinal value of crocin has attracted much attention. Crocin II is the indicative active ingredient in crocin. It has been proven that crocin II has calcium antagonism, hypoglycemic and lipid-lowering effects, anti-atherosclerosis, liver protection, anti-tumor, and treatment of coronary heart disease.

[0005] However, with respect to septic renal injury, there are no relevant patents and non-patent literatures reporting that crocin II has an improving effect on septic renal injury. Summary of the invention

[0006] The purpose of the present invention is to solve the deficiencies of the prior art, to realize the development and utilization of natural products using modern drug research methods, and to provide the use of crocin II in the preparation of drugs for preventing or treating septic renal injury in combination with a large number of pharmacodynamic experimental screenings.

[0007] To achieve the purpose of the present invention, the present invention adopts the following technical solutions:

[0008] The present invention provides use of crocin II in preparing a medicine for preventing or treating sepsis-induced renal injury.

[0009] The present invention also provides the use of crocin II as the sole active ingredient in preparing a medicine for preventing or treating sepsis-induced kidney injury.

[0010] As an alternative, in the above use, the septic renal injury is septic acute renal injury.

[0011] As an alternative, in the above use, the renal injury includes renal tissue oxidative stress, renal tissue injury, renal edema, renal tissue inflammation and glomerular injury.

[0012] As an optional mode, in the above-mentioned use, the medicine further comprises a pharmaceutically acceptable carrier.

[0013] As an optional mode, in the above-mentioned use, the dosage form of the drug is an oral dosage form.

[0014] As an optional mode, in the above-mentioned use, the oral dosage form is a tablet, a capsule, a granule or an oral liquid.

[0015] The above-mentioned various dosage forms can be prepared according to conventional processes in the field of pharmaceutical preparations.

[0016] Crocin II can be extracted and separated from plants containing these active ingredients such as saffron, gardenia, burdock, mimosa, etc. by conventional extraction and purification methods in the field of natural medicinal chemistry, or it can be purchased from commercial products.

[0017] In the medical uses described above, the administration method, administration time, administration times and administration frequency of Crocin II of the present invention need to be determined according to the specific diagnosis results of the disease, which is within the technical scope mastered by those skilled in the art.

[0018] The treatment regimen for mice is applied to humans, and the effective dose of all drugs for humans can be converted by the effective dose of the drug for mice, which is also easy to achieve for those of ordinary skill in the art. It is understood by those skilled in the art that the actual dosage may be lower than the converted dosage range. The effective amount and specific treatment regimen for a subject may be affected by many factors, including the age, weight, sex, diet, administration time, disease susceptibility, disease progression, doctor's judgment, etc. of the subject.

[0019] In order to better understand the essence of the present invention, the pharmacodynamics experiments and their results are used in the following specific implementation section to further illustrate the new use of crocin II of the present invention in the pharmaceutical field.

[0020] Through the above technical solution, the present invention has at least the following advantages:

[0021] (1) The present invention constructs a sepsis model to conduct experimental research on crocin II in preventing or treating renal damage caused by sepsis. The results show that crocin II has a good protective effect on the sepsis multi-organ injury model induced by cecal ligation and puncture (CLP) with small side effects, and can be used to prepare protective drugs for renal damage caused by sepsis.

[0022] (2) The present invention provides the use of crocin II in the preparation of a therapeutic drug for kidney damage caused by sepsis, which has significant effects in improving acute kidney damage and exerting anti-inflammatory effects with few side effects, and has considerable clinical application prospects and commercial value.

[0023] (3) The present invention provides more possibilities for targeted drug treatment of sepsis-induced renal injury. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 : H&E staining results of renal tissue damage in mice in each group. A: sham operation group; B: model group; C: crocin II group; D: crocetin group; D: dexamethasone group, magnification 40x. DETAILED DESCRIPTION

[0025] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0026] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0027] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0028] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present application description and examples are exemplary only.

[0029] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0030] The present invention will be further described with reference to specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the scope of the present invention.

[0031] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0032] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.

[0033] Example:

[0034] The present invention studies the use of crocin II in the preparation of a drug for preventing or treating sepsis-induced renal injury through animal experiments. The present invention uses C57BL / 6 mice as experimental subjects, simulates a sepsis-induced renal injury model through cecal ligation and puncture (CLP), and administers crocin II orally or intragastrically, thereby exploring the application of crocin II in protecting renal injury caused by sepsis. 1. Experimental method:

[0035] 1.1 Animal husbandry:

[0036] Six-week-old SPF male C57BL / 6 mice (purchased from Liaoning Changsheng Biotechnology Co., Ltd.) weighing 20-22 g were selected. The experiment was carried out after a one-week adaptation period. Six mice were placed in a cage, and standard mouse feed and normal temperature drinking water that had been strictly sterilized were regularly replaced.

[0037] 1.2 Animal modeling:

[0038] The surgical area of ​​the mouse abdomen was prepared, and 75% ethanol was used to disinfect it repeatedly for 3 times. A longitudinal incision of about 1 cm was made in the middle and left part of the mouse abdomen, and the skin, fascia, and muscle were cut in layers to expose the abdominal cavity. With blunt-tipped ophthalmic forceps in both hands, the abdominal cavity was gently explored to find and free the mouse's cecum, which was ligated about 1 cm away from the end of the cecum. A 25-gauge needle was used to puncture the end of the ligated cecum. A blunt-tipped ophthalmic forceps was used to gently squeeze out a small amount of intestinal contents from the puncture site and then return the cecum. The surgical incision was sutured and closed in layers in the opposite order of the laparotomy. 37°C water bath with normal saline and 1 mL was injected subcutaneously into the back neck of the postoperative mouse for resuscitation. The mouse was placed in a supine position in a cage and kept warm. Tissues were obtained after 5 days. By observing the survival status of the mice and detecting the pathological damage of the organs and the levels of pro-inflammatory cytokines in the mice in subsequent experiments, it was proved that the CLP-induced sepsis mouse model was successfully constructed.

[0039] 1.3 Animal administration:

[0040] Crocin II and crocetin (purchased from Shenyang Sandu Pharmaceutical Technology Co., Ltd.) were dissolved in a solvent containing 4% DMSO, 40% PEG400 and 56% normal saline.

[0041] The animal experiment was divided into 5 groups: sham operation group, model group, crocin II group, crocetin group, and dexamethasone group, with 12 mice in each group.

[0042] Dosing began 6 hours after surgery.

[0043] The mice in the sham operation group and the model group were intragastrically administered with a solvent containing 4% DMSO, 40% PEG400 and 56% saline every day for 5 consecutive days.

[0044] For the mice in the crocin II group and the crocetin group, 40 mg / kg of crocin II and crocetin were administered orally to the mice every day for 5 consecutive days.

[0045] The mice in the dexamethasone group were intraperitoneally injected with 2 mg / kg diluted with normal saline every day for 5 consecutive days.

[0046] 1.4 Testing indicators:

[0047] (1) Survival rate of mice in each group at the end of the experiment on day 5.

[0048] (2) At the end of the experiment on the 5th day, blood was collected from the retrobulbar vein of each group of mice and centrifuged at 3000 rpm for 10 min. The supernatant was collected and the blood urea nitrogen (BUN), blood creatinine (CRE), TNF-α, and IL-6 levels were measured using a kit.

[0049] (3) Histopathological examination: At the end of the experiment on the 5th day, the kidney tissues of the mice were removed, fixed with 4% paraformaldehyde, embedded in paraffin and sectioned. The pathological changes of the kidney tissues were observed under an optical microscope after HE staining.

[0050] 1.5 Data representation and statistical analysis methods:

[0051] The results were analyzed using SPSS 27.0 statistical software, and the data were expressed as mean ± standard deviation. One-way analysis of variance was used for comparison among multiple groups, and t-test was used for comparison between two groups. P < 0.05 indicated statistically significant differences.

[0052] 2. Experimental results:

[0053] 2.1 Survival rate of mice in each group:

[0054] There were 12 mice in each group. At the end of the experiment on the 5th day, the survival rate of mice in each group was shown in Table 1 below.

[0055] Table 1: Survival rate of mice in each group (%)

[0056]

[0057]

[0058] The experimental results showed that all mice in the sham operation group survived at the end of the experiment. The survival rate of mice in the model group was only 25%, and the survival rate of mice in the crocin II group was significantly improved to 75%, close to the dexamethasone group (83.33%). However, the survival rate of the crocetin group was lower (33.33%), only slightly higher than that of the model group.

[0059] 2.2 Renal function test results of mice in each group:

[0060] At the end of the experiment on the 5th day, the renal function test results of mice in each group are shown in Table 2 below.

[0061] Table 2: Renal function test results of mice in each group

[0062] Group BUN (mmol / L) CRE (μmol / L) Sham operation group 4.82±0.95 15.78±4.35 Model Group <![CDATA[16.37±8.58 *** ]]> <![CDATA[58.72±19.84 *** ]]> Crocin II group <![CDATA[8.13±2.14 ## ]]> <![CDATA[25.65±5.92 *,## ]]> Crocetin group <![CDATA[14.18±6.71 *** ]]> <![CDATA[51.94±15.66 *** ]]> Dexamethasone group <![CDATA[6.55±1.23 ## ]]> <![CDATA[21.56±5.17 ## ]]>

[0063] Note: Compared with the sham operation group, * p<0.05, *** p<0.001; compared with the model group, ## p<0.01.

[0064] The experimental results showed that at the end of the experiment, the BUN and CRE levels of the model group mice were significantly higher than those of the sham operation group, indicating that the model was successfully established. Crocin II can significantly reduce the BUN and CRE levels of CLP-induced septic mice ( ## p<0.05), and the degree of improvement of the above two indicators was comparable to that of dexamethasone, indicating that crocin can significantly improve the renal function of septic mice. However, crocin did not basically reduce the BUN and CRE of CLP-induced septic mice, and these two indicators of this group were only slightly better than those of the model group.

[0065] 2.3 Levels of TNF-α and IL-6 in serum of mice in each group:

[0066] At the end of the experiment on the 5th day, the levels of serum TNF-α and IL-6 in each group of mice are shown in Table 3 below.

[0067] Table 3: Serum TNF-α and IL-6 levels in each group of mice

[0068]

[0069]

[0070] Note: Compared with the sham operation group, * p<0.05, ** p<0.01, *** p<0.001; compared with the model group,## p<0.01, ### p<0.001.

[0071] The experimental results showed that at the end of the experiment, the levels of TNF-α and IL-6 in the model group mice were significantly higher than those in the sham operation group, indicating that the model was successfully established. Crocin II can significantly reduce the levels of TNF-α and IL-6 in CLP-induced sepsis mice ( ## p<0.05), but its improvement on the above two inflammatory indicators is not as good as that of dexamethasone, which indicates that crocin can significantly reduce the inflammatory indicators of mice with septic renal injury, and the above results may also reflect from the side that the improvement of renal function may not only be due to the anti-inflammatory effect of crocin II. From the above results, it can also be seen that crocetin also has a certain anti-inflammatory effect, but it will not improve the renal function indicators of mice with septic renal injury, which also reflects that the improvement of renal function is not necessarily clearly related to inflammatory indicators.

[0072] 2.4 Results of renal tissue damage in each group of mice:

[0073] The results of H&E staining of renal tissue damage in each group of mice are shown in Figure 1 As shown in the figure, compared with the sham operation group, the model group showed renal edema, renal tissue inflammation and glomerular damage, and tubular epithelial cell necrosis, increased vacuoles, and increased cell fragments were observed, accompanied by glomerular congestion and necrosis. Dexamethasone and crocin II can significantly improve the above-mentioned renal tissue pathological damage in CLP-induced septic mice. However, the results of renal tissue pathological damage in the crocetin group were similar to those in the model group, indicating that crocetin cannot improve renal tissue damage in CLP-induced septic mice.

[0074] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. Use of crocin II in the preparation of a drug for preventing or treating septic renal injury.

2. Use of crocin II as the sole active ingredient in the preparation of a drug for preventing or treating septic renal injury.

3. The use according to claim 1 or claim 2, characterized in that: The septic renal injury is septic acute renal injury.

4. The use according to claim 1 or claim 2, characterized in that: The renal injury includes renal tissue oxidative stress, renal tissue injury, renal edema, renal tissue inflammation and glomerular injury.

5. The use according to claim 1 or claim 2, characterized in that: The medicament further comprises a pharmaceutically acceptable carrier.

6. The use according to claim 5, characterized in that: The dosage form of the drug is an oral dosage form.

7. The use according to claim 6, characterized in that: The oral dosage form is tablet, capsule, granule or oral liquid.

Images