Application of Bamboo Leaf Orchid and its extract in preparing medicine for treating kidney injury

Abstract

The present invention provides the use of Bamboo Leaf Orchid and its extract in the preparation of a drug for treating kidney injury, and discloses a method for extracting the Bamboo Leaf Orchid extract. The present invention first demonstrates, through experiments in biological kidney injury models, that Bamboo Leaf Orchid and its extract have significant protective effects against acute kidney injury in mice and chronic kidney injury in rats. This provides a new application for the traditional Chinese medicine Bamboo Leaf Orchid and offers new insights into the development of novel anti-kidney injury drugs.

Description

Technical Field

[0001] The invention relates to application of bamboo leaf orchid and an extract thereof, in particular to application in preparing medicine for treating kidney damage. Background Art

[0002] The kidney is a complex biological system with diverse regulatory mechanisms and physiological functions. The "Kidney Disease: Improving Global Outpatient Clinical Practice Guidelines" states that when renal function is impaired or gradually declines, the kidney's excretion and regulatory functions are also reduced. Severe renal impairment can lead to life-threatening uremia. Epidemiological studies have shown that approximately 15% of adults and 25% of hospitalized children experience acute kidney injury, and the incidence of acute kidney injury is even higher among patients receiving intensive care (>57% in one large international study). Multiple studies have shown that acute kidney injury increases the risk of chronic kidney disease, leading to fibrosis, vascular rarefaction, tubular loss, glomerular sclerosis, and chronic inflammation after the acute phase of injury. If left untreated, it can gradually develop into chronic renal failure, severely impacting quality of life. Despite decades of intensive research, the complex pathogenesis of kidney injury remains unclear.

[0003] Currently, there is no effective treatment for kidney damage in clinical practice, and targeted drugs have difficulty achieving synergistic effects. Mainstream clinical drugs such as hormones, antibiotics, and antihypertensive drugs require long-term use and high concentrations. However, high doses and high concentrations often lead to serious adverse reactions. Nonsteroidal anti-inflammatory drugs (NSAIDs), hormones, penicillins, ACE inhibitors, immunosuppressants, etc. often cause serious side effects. Therefore, improving the safety and effectiveness of drugs for treating kidney disease is of great significance.

[0004] Bamboo orchid (scientific name: Arundina graminifolia (D.Don) Hochr.), also known as Baiyangjie, belongs to the genus Bamboo orchid of the Orchidaceae family. It is a commonly used medicine of the Dai people and is used to treat food poisoning. As a traditional Chinese medicine for detoxification, the application of Bamboo orchid and its extracts in other new fields has also been discovered one after another. For example, the Chinese patent with announcement number CN107050301B applies it to inhibit the growth of Salmonella; the Chinese patent with publication number CN105326890A applies it to the preparation of drugs for treating gastric ulcers. The inventors of the present invention found in the process of drug activity screening that Bamboo orchid and its extracts have a good effect on resisting kidney damage, while no records of Bamboo orchid being used for resisting kidney damage were found in the prior art. The application of Bamboo orchid and its extracts will help provide ideas for new anti-kidney damage drugs. Summary of the Invention

[0005] Based on the above background technology, the present invention provides the use of Bamboo Leaf Orchid and its extract in the preparation of anti-kidney injury drugs.

[0006] In the above application, the Bamboo Herb extract is a Bamboo Herb water extract or an alcohol extract.

[0007] In the above application, the extraction method of the Bamboo Leaf Orchid extract is:

[0008] Step 1: Weigh 1 part of Bamboo Leaf Orchid, add 5-20 parts of extraction solvent, and filter to obtain the extract;

[0009] Step 2: Filter the extract obtained in step 1, concentrate and dry to obtain the bamboo orchid extract.

[0010] In the above application, the extraction solvent is methanol, the extraction times are 2-3 times, and each extraction takes 0.5-2 hours.

[0011] In the above application, the extraction method of the extract in step 1 is ultrasonic extraction, and the extraction temperature is 40-60°C.

[0012] In the above application, the concentration in step 2 is rotary evaporation concentration, and the rotary evaporation temperature is 50°C.

[0013] In the above application, the drying in step 2 is drying, decompression or freeze-drying.

[0014] In the above application, the crude drug amount of the bamboo orchid or bamboo extract is 0.1-1g per kilogram of body weight (human).

[0015] In the above application, the renal injury is acute renal injury caused by cisplatin or chronic renal injury caused by adenine.

[0016] In the above application, the dosage form of the drug is tablets, granules or oral dosage forms.

[0017] During the drug activity screening process, the inventors of the present invention confirmed through pharmacological experiments that Bamboo Leaf Orchid and its extract can inhibit kidney inflammatory factors and effectively protect against acute kidney injury caused by cisplatin and chronic kidney injury caused by adenine. This provides a new application for the traditional Chinese medicine Bamboo Leaf Orchid and also provides new ideas for new anti-kidney injury drugs. In addition, the Bamboo Leaf Orchid extraction method provided in the examples is simple in process and has low extraction difficulty, making it easy to promote its application in anti-kidney injury drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 These are HE-stained light microscopic images of kidney tissues of mice in the blank group, model group, high / medium / low dose groups in Example 2 of the present invention; ##p<0.01 compared with the blank group, ***p<0.001, **p<0.01, *p<0.05 compared with the model group. DETAILED DESCRIPTION

[0019] The present invention is further described below with reference to the accompanying drawings and specific embodiments.

[0020] Example 1: Preparation of Bamboo Herb Extract.

[0021] The extraction method of Bamboo Leaf Orchid extract is:

[0022] Step 1: Weigh 1 part of Bamboo Leaf Orchid, add 5-20 parts of methanol, reflux, ultrasonicate, or extract multiple times, filter, and combine to obtain an extract. In this embodiment, water bath ultrasonic extraction is performed 2-3 times at an extraction temperature of 40-60°C. The effective amount of Bamboo Leaf Orchid or its extract that can exert an anti-nephritis effect is 0.1-1g of the raw drug per kilogram of body weight (human).

[0023] Step 2: Filter the extract obtained in step 1, evaporating and concentrating at 50°C until the extract stops clinging to the wall. Dry, decompress, or freeze-dry the filtrate to obtain the Bamboo Herb Extract. Weigh the extract and calculate the extraction yield.

[0024] Example 2: Experimental study on the pharmacodynamic activity of Bamboo Herb Extract against acute kidney injury induced by cisplatin.

[0025] a. Modeling and drug administration methods:

[0026] 50 KM mice were selected and divided into a blank group, a model group, and a high / medium / low dose group. The blank group was not subjected to modeling. The groups other than the blank group were prepared with pure water to prepare a 1 mg / mL cisplatin concentration, and the model was established by a single intraperitoneal injection of 10 mg / kg. The high, medium, and low dose groups were administered with the Bamboo Leaf Extract prepared in Example 1, and the dosage was equivalent to 6 / 3 / 1.5 g of Bamboo Leaf Extract per kilogram of body weight (mouse) per day, respectively. Modeling was established 14 days after pre-oral administration, and the drug was continued for 3 days. On the 3rd day, blood was collected by removing the eyeball 2 hours after administration, and the kidney was taken for preservation and sample retention. The slicing process was as follows:

[0027] Kidney tissue was fixed with 4% neutral formaldehyde solution and stored at 4°C for 12 hours. After fixation, the kidney tissue was removed, rinsed in running water for 1 hour, and dehydrated according to the following procedure: 75% ethanol for 1 hour; 85% ethanol for 1 hour; 95% ethanol for 1 hour; 95% ethanol for 3 hours; anhydrous ethanol for 0.5 hours; anhydrous ethanol for 2 hours; xylene I for 1 hour; xylene II for permeabilization for 1 hour; paraffin I for 1 hour; and paraffin II for 3 hours. Then embed in paraffin, slice into 3-4 microns and bake at 60℃ for 5 hours, dewax in xylene five times, each time for 5 minutes, then place in anhydrous ethanol I for 5 minutes; place in anhydrous ethanol II for 5 minutes; place in 95% ethanol I for 5 minutes; place in 95% ethanol II for 5 minutes; place in 80% ethanol for 5 minutes; place in 70% ethanol for 5 minutes; rinse with running water for 5 minutes; place in hematoxylin fuel for 5 minutes; color separation with 0.5% hydrochloric acid alcohol for 5 seconds; wash with water for blueing for 20 minutes; place in 70% ethanol for 5 minutes; place in 80% ethanol for 5 minutes; place in eosin dye solution for 20 seconds; place in 95% ethanol I for 5 minutes; place in 95% ethanol II for 5 minutes; place in xylene + ethanol 1:1 solution for 5 minutes; place in xylene I for 5 minutes; place in xylene II for 5 minutes; place in xylene III for 5 minutes; after staining, dry and seal with neutral resin, and observe the pathological changes of renal tissue under a microscope.

[0028] b. Implementation results:

[0029] The experimental results are as follows Figure 1 As shown in AD, Figure 1 A is a comparison of kidney samples from 5 groups of mice. The renal tissue morphology of mice in the blank group was normal; the renal tubular epithelial cells of mice in the model group were edematous and necrotic, with inflammatory cell infiltration in the renal interstitium, and the glomerular morphology was generally normal; the degree of renal tubular necrosis in the high / medium / low dose groups of Bamboo Leaf Orchid was significantly reduced compared with the model group, and the glomeruli were generally normal.

[0030] Figure 1 BD displayed the organ index, urea nitrogen, and blood creatinine of 5 groups of mice respectively. The results showed that the high / medium / low dose groups of Bamboo Leaf Orchid could effectively reduce the kidney organ index of mice caused by cisplatin, and reduce the content of urea nitrogen and blood creatinine in the model group, indicating that Bamboo Leaf Orchid extract has a certain protective effect on cisplatin-induced acute kidney injury in mice.

[0031] Example 3: Pharmacological activity test of Bamboo Herb Extract on chronic kidney damage caused by adenine.

[0032] a. Modeling and drug administration methods:

[0033] Sixty SD rats were selected and divided into blank group, model group, high / medium / low dose group and positive drug group. The blank group was not subjected to modeling treatment. Except for the 10 rats in the blank group, the other groups were gavaged with adenine at a dose of (300 mg / kg / d) and a suspension prepared with distilled water at a fixed time every day for 4 consecutive weeks. The rats were allowed to drink water and eat freely to form a chronic renal injury model in rats.

[0034] After modeling, rats were assigned to high, medium, and low doses of Bamboo Leaf Herb at a daily dose of 3, 2, and 1 g per kilogram of body weight, respectively. Dosing began one day after modeling and continued for 28 days. The blank and model groups were gavaged with the solvent, the Bamboo Leaf Herb group was gavaged with the Bamboo Leaf Herb extract described in Example 1, and the positive drug group was gavaged with losartan.

[0035] b. Experimental results:

[0036] The comparison of organ indexes of each group is as follows:

[0037]

[0038]

[0039] Compared with the blank group##p<0.01; compared with the model group**p<0.01, *p<0.05.

[0040] The comparison of creatinine and urea nitrogen content in each group is as follows:

[0041]

[0042] Compared with the blank group##p<0.01; compared with the model group**p<0.01, *p<0.05.

[0043] The comparison of TNF-α, IL-1β, and IL-6 levels in each group is as follows:

[0044]

[0045] Compared with the blank group##p<0.01; compared with the model group**p<0.01, *p<0.05.

[0046] The above results show that bamboo orchid extract can effectively inhibit kidney inflammatory factors and protect chronic kidney damage caused by adenine or acute kidney damage caused by cisplatin.

[0047] The preferred embodiments of the present invention have been specifically described above, but the present invention is not limited to the described embodiments. Those skilled in the art may make various equivalent modifications or substitutions without departing from the spirit of the present invention. These equivalent modifications or substitutions are all included in the scope defined by the claims of this application.

Claims

1. The use of bamboo orchid extract in the preparation of a drug for treating chronic kidney damage caused by adenine, wherein the bamboo orchid extract is bamboo orchid methanol extract, the extraction method is ultrasonic extraction, and the extraction temperature is 40~60℃.

2. The use according to claim 1, characterized in that: The extraction method of the Bamboo Herb extract is: Step 1: Weigh 1 part of Bamboo Leaf Orchid, add 5-20 parts of methanol, perform ultrasonic extraction, and filter to obtain the extract. Step 2: Filter the extract obtained in step 1, concentrate and dry to obtain the bamboo orchid extract.

3. The use according to claim 2, characterized in that: The extraction times are 2-3 times, each extraction time is 0.5-2h.

4. The use according to claim 2, wherein: The concentration in step 2 is carried out by rotary evaporation, and the rotary evaporation temperature is 50°C.

5. The use according to claim 2, characterized in that: The drying in step 2 is oven drying, reduced pressure drying or freeze drying.

6. The use according to claim 1, characterized in that: The dosage form of the medicine is tablets or granules.

Images