阅读说明：本技术 柠檬苦素在制备预防和治疗急性肾损伤药物中的用途 (Application of limonin in preparation of medicine for preventing and treating acute kidney injury ) 是由 傅海燕 周栋 刘史佳 周先可 卢嘉微 于 2019-10-09 设计创作，主要内容包括：本发明属于医药领域,涉及一种中草药单体柠檬苦素在制备预防和治疗急性肾损伤药物中的用途。本发明分别通过体内、体外研究发现,柠檬苦素具有明显的抑制肾小管上皮细胞凋亡,促进增殖的生物学作用,其对缺血-再灌注或化疗药物顺铂诱导的急性肾损伤具有明显的肾保护作用。因此,柠檬苦素可以应用于制备预防和治疗急性肾损伤的药物,具有明显的肾保护作用,且毒副作用小,成本低廉。(The invention belongs to the field of medicines, and relates to application of a Chinese herbal medicine monomer limonin in preparation of a medicine for preventing and treating acute kidney injury. The limonin is found to have obvious biological effects of inhibiting the apoptosis of renal tubular epithelial cells and promoting proliferation through in vivo and in vitro researches respectively, and has obvious renal protection effect on acute renal injury induced by ischemia-reperfusion or chemotherapeutic drug cisplatin. Therefore, the limonin can be applied to preparing the medicine for preventing and treating acute kidney injury, has obvious kidney protection effect, and has small toxic and side effect and low cost.)

1. Application of limonin in preparing medicine for preventing and treating acute kidney injury is provided.

2. Use according to claim 1, characterized in that: the acute kidney injury is caused by ischemia-reperfusion or chemotherapy drug cisplatin.

3. Use according to claim 1, characterized in that: the medicine is used for inhibiting the renal tubular epithelial cell apoptosis caused by ischemia-reperfusion or chemotherapy medicine cisplatin.

4. Use according to claim 1, characterized in that: the medicine is used for promoting the proliferation of the renal tubular epithelial cells caused by the ischemia-reperfusion or chemotherapy medicine cisplatin.

Technical Field

The invention belongs to the field of medicines, relates to application of a Chinese herbal medicine monomer Limonin (Limonin), and particularly relates to application of the Limonin in preparation of a medicine for preventing and treating acute kidney injury.

Background

Acute Kidney Injury (AKI) is a common critical clinical condition, has the disadvantages of Acute onset, rapid progression and high mortality, and seriously threatens human health. The patients mainly show rapid decline of kidney function and accumulation of metabolic waste products, and the clinical diagnosis is based on increase of Serum Creatinine (Scr) and decrease of urine volume. The incidence of AKI is high and tends to increase year by year. Epidemiological data show: even mild, reversible AKI causes permanent damage to kidney tissue; severe AKI leads to an irreversible decrease in kidney function, increasing the risk of death. Due to the lack of targeted prevention and protection drugs at present, AKI and chronic kidney disease caused by further development of AKI are still a great problem in China and the global medical health industry. Therefore, it is very urgent and important to deeply explore the pathogenesis of AKI, search for specific therapeutic targets, and develop effective preventive and therapeutic drugs.

Although mechanisms of AKI caused by various causes such as renal ischemia, nephrotoxic drugs, sepsis, urinary obstruction and the like are different, the AKI basically relates to several pathophysiological links such as change of hemodynamics, oxidative stress injury and inflammatory reaction. The above pathophysiological changes all cause damage of renal tubular epithelial cells, glomerular filtration and renal tubular reabsorption disorders, rapid decline of renal function, further aggravation of renal damage, and finally irreversible decline of renal function. After AKI occurs, renal tubular epithelial cells are firstly affected, and a large number of cells enter a G1 phase from a G0 phase to repair injury; if the repair is successful, entering the next cell cycle; if the repair fails or the damage is too severe, apoptosis and necrosis occur, inflammatory factors are released, inflammatory reaction of tissues is mediated, reaction is amplified, and the damage is aggravated. Therefore, how to inhibit tubular cell apoptosis and promote tissue repair is the key point for effectively preventing and treating the occurrence and development of AKI.

Limonin (Limonin) is a triterpenoid present in rutaceae and meliaceae plants, an important secondary metabolite with high biological activity in plants. In recent years, limonin is receiving more and more attention from scientists, and has been found to have good anti-inflammatory, antibacterial, analgesic, antitumor and other effects. In a liver injury model caused by rat ischemia-reperfusion, limonin can reduce oxidative stress level and alleviate hepatocyte degeneration necrosis by inhibiting lactate dehydrogenase activity; the limonin can also inhibit activation of a TLR-4-NF kappa B signal channel, inhibit release of inflammatory factors and play a role in protecting the liver. In addition, in vivo and in vitro researches on mice such as colon cancer, breast cancer, liver cancer and the like also find that the limonin can inhibit the proliferation of tumor cells, promote the apoptosis of the tumor cells and play an effective anticancer role. However, no studies on limonin in the field of acute kidney injury treatment have been reported at home and abroad.

Disclosure of Invention

The invention aims to provide a new application of a traditional Chinese medicine monomer Limonin (Limonin), in particular to an application of the Limonin in preparing a medicine for preventing and treating acute kidney injury.

According to a further feature of the use of the present invention, the acute kidney injury is acute kidney injury caused by ischemia-reperfusion or a chemotherapeutic drug cisplatin.

according to a further feature of the use of the invention, the medicament is a medicament for inhibiting apoptosis of renal tubular epithelial cells caused by cisplatin, an ischemia-reperfusion or chemotherapeutic drug.

According to a further characteristic of the use according to the invention, the said drug is a drug for promoting the proliferation of tubular epithelial cells induced by the ischemia-reperfusion or chemotherapeutic drug cisplatin.

The invention discovers that limonin can promote the proliferation of renal tubular epithelial cells by inhibiting the apoptosis of the renal tubular epithelial cells and plays a remarkable role in protecting renal function through in vivo zoology experiments and in vitro cytology experiments respectively. Therefore, the limonin can be applied to preparing the medicine for preventing and treating acute kidney injury, has obvious kidney protection effect, and has small toxic and side effect and low cost.

Specifically, the inventors found in the mouse bilateral ischemia-reperfusion (BIRI) and cisplatin-induced AKI models, respectively: after the mouse is treated by limonin by gastric lavage, the serum creatinine and the serum urea nitrogen level can be obviously restored, the pathological injury of the kidney tissue is reduced, and the apoptosis of the renal tubular epithelial cells is reduced. In vitro cytology experiments further prove that the limonin has obvious inhibition effects on renal tubular epithelial cell apoptosis induced by hypoxia-reoxygenation and cisplatin respectively; meanwhile, limonin can promote proliferation of renal tubular epithelial cells. In conclusion, limonin has a significant kidney protection effect and no significant toxic or side effects, and therefore can be used for preparing effective AKI prevention and treatment medicines.

the invention discloses for the first time: limonin has obvious protective effect on acute kidney injury of mice caused by ischemia-reperfusion and cisplatin injection. The research is not reported at home and abroad, and the achievement of the research has great practical significance for developing AKI prevention and treatment medicines.

Drawings

Figures 1A-1C show that limonin can restore acute renal function decline and renal tissue injury caused by bilateral ischemia-reperfusion (BIRI) injury of the mouse kidney. Wherein, FIG. 1A shows the biochemical method for detecting the serum creatinine level change of mice; FIG. 1B shows the biochemical detection of changes in serum urea nitrogen levels in mice; FIG. 1C and FIG. 1D show PAS (Periodic Acid-Schiff stain) staining showing histopathological changes in mouse kidney.

FIGS. 2A to 2E show that limonin inhibits apoptosis of kidney tissue caused by bilateral ischemia-reperfusion (BIRI) injury of mouse kidney, and expression change of apoptosis molecules is detected by using Western immunoblotting (WB) method.

FIGS. 3A to 3F show that limonin inhibits renal tubular epithelial cell apoptosis caused by hypoxia-reoxygenation in vitro, expression change of apoptosis molecules is detected by a Western Blot method, and apoptotic cell proportion is detected by a flow cytometry.

FIGS. 4A to 4D show that limonin restores acute renal function decline and renal tissue injury in mice induced by the chemotherapeutic cisplatin; wherein, FIG. 4A shows the biochemical method for detecting the serum creatinine level change of the mouse; FIG. 4B shows the biochemical detection of changes in serum urea nitrogen levels in mice; figure 4D shows PAS staining showing histopathological changes in mouse kidney.

FIGS. 5A to 5E show that limonin inhibits cisplatin-induced apoptosis of mouse kidney tissue cells, and changes in expression of apoptotic molecules are detected by Western Blot.

FIGS. 6A-6E show that limonin inhibits cisplatin-induced tubular epithelial cell apoptosis in vitro; detecting the expression change of apoptosis molecules by using a Western Blot method; the proportion of apoptotic cells was detected by flow cytometry.

FIGS. 7A to 7D show that limonin promotes proliferation of renal tubular epithelial cells in vitro, and expression changes of cells proliferation related molecules are detected by a Western Blot method.

Detailed Description

The invention will now be further described, by way of example only, with reference to the accompanying drawings.