阅读说明：本技术 甘草次酸在制备治疗肾小管间质炎症药物中的应用 (Application of glycyrrhetinic acid in preparation of medicine for treating tubulointerstitial inflammation ) 是由 许艳芳 陈志敏 赖坤美 叶铿 于 2021-02-04 设计创作，主要内容包括：本申请属于医药技术领域,具体涉及甘草次酸在制备治疗肾小管间质炎症药物中的应用,所述肾小管间质炎症为高尿酸引起的肾小管间质炎症。甘草次酸通过抑制高尿酸引起的肾小管上皮细胞坏死、抑制高尿酸晶体引起的单核巨噬细胞Nlrp3炎症小体的激活、GSDMD介导的细胞焦亡和炎症因子的分泌,抑制高尿酸引起肾小管间质炎症。(The application belongs to the technical field of medicines, and particularly relates to application of glycyrrhetinic acid in preparation of a medicine for treating tubulointerstitial inflammation, wherein the tubulointerstitial inflammation is caused by high uric acid. Glycyrrhetinic acid can inhibit renal tubular interstitial inflammation caused by hyperuricemia by inhibiting renal tubular epithelial cell necrosis caused by hyperuricemia, inhibiting activation of mononuclear macrophage Nlrp3 inflammasome caused by hyperuricemia crystal, and cell apoptosis and inflammatory factor secretion mediated by GSDMD.)

1. Use of glycyrrhetinic acid in the preparation of a medicament for treating tubulointerstitial inflammation, wherein the tubulointerstitial inflammation is caused by hyperuricemia.

2. The use of claim 1, wherein glycyrrhetinic acid reduces renal function impairment of hyperuricemic nephropathy and lowers blood urea nitrogen and creatinine.

3. The use of claim 1, wherein glycyrrhetinic acid reduces uric acid-induced renal tubular injury and interstitial inflammatory changes.

4. The use of claim 1, wherein glycyrrhetinic acid reduces neutrophil infiltration in hyperuricemic kidney tissue.

5. The use of claim 1, wherein glycyrrhetinic acid reduces hyperuricemic renal tissue macrophage infiltration.

6. The use of claim 1, wherein glycyrrhetinic acid reduces sodium urate crystal-induced renal tubular epithelial cell death.

7. Use according to claim 1, characterized in that glycyrrhetinic acid reduces the sodium urate crystal-induced activation of the macrophage NLRP3 inflammasome, the shearing of the cell pyroptosis marker protein GSDMD and the secretion of IL-1 β.

8. Use of a composition comprising glycyrrhetinic acid for the manufacture of a medicament for the treatment of tubulointerstitial inflammation, wherein the tubulointerstitial inflammation is caused by hyperuricemia.

9. The use of claim 8, wherein the composition is a medicament comprising glycyrrhetinic acid as an active ingredient, in combination with pharmaceutically acceptable excipients.

Technical Field

The application belongs to the technical field of medicines, and particularly relates to application of glycyrrhetinic acid in preparation of a medicine for treating tubulointerstitial inflammation, wherein the tubulointerstitial inflammation is caused by high uric acid.

Background

In recent years, the prevalence rate of hyperuricemia is on the rise, the incidence rate of hyperuricemic nephropathy is gradually obvious, and when the concentration of blood uric acid exceeds 6.8mg/dL, urate can be deposited in kidney tissues to form mono-sodiumcurate crystals and cause interstitial inflammatory reaction of renal tubules, and the pathology is characterized by the death of renal tubular epithelial cells and the infiltration of interstitial inflammatory cells. The adenine diet-induced kidney injury of mice simulates hyperuricemic nephropathy, and a large amount of inflammatory cell infiltration can be obviously seen in renal interstitium, which is mainly neutrophilic granulocyte and macrophage, while the pathological changes of glomeruli are not obvious, and the special characteristics are different from diabetic nephropathy which is mainly characterized by glomerular disease.

Glycyrrhetinic acid is a pentacyclic triterpenoid compound, has the effects of corticoid hormone, strong anti-inflammation, immunity regulation and cell death inhibition by inhibiting DNA damage, has a protective effect in researches such as diabetic nephropathy and radiation lung injury, but has no report on renal tubulointerstitial inflammation caused by high uric acid.

Patent CN106539803A discloses the use of 18 β -glycyrrhetinic acid (or salt), one of the main components in glycyrrhiza, as an adjuvant treatment drug for alleviating acute kidney injury caused by cisplatin, a chemotherapeutic drug. Cisplatin, as a common chemotherapeutic drug, has nephrotoxicity that limits its clinical application. The 18 beta-glycyrrhetinic acid can inhibit the levels of creatinine and urea nitrogen in acute kidney injury induced by cisplatin, reduce the expression of KIM-1 injury protein and inhibit the apoptosis of renal tubular epithelial cells caused by the cisplatin. However, the effect of glycyrrhetinic acid on the inflammation of the tubulointerstitial inflammation caused by hyperuricemia is not disclosed.

Patent CN102813661B discloses a glycyrrhetinic acid derivative which has a good kidney injury protection effect and can be used for preparing a medicament for preventing and treating kidney injury diseases; the glycyrrhetinic acid derivative can reduce blood urea nitrogen and creatinine, and improve renal function. Nor does the document mention that glycyrrhetinic acid is effective in causing tubulointerstitial inflammation by hyperuricemia.

Disclosure of Invention

The inventor has surprisingly found that glycyrrhetinic acid has the effect of obviously relieving the renal tubular interstitial inflammation caused by hyperuricemia.

The invention aims to provide application of glycyrrhetinic acid in preparing a medicine for treating tubulointerstitial inflammation, wherein the tubulointerstitial inflammation is caused by high uric acid.

The glycyrrhetinic acid can relieve renal function damage of hyperuricemia nephropathy, and reduce blood urea nitrogen and creatinine.

The glycyrrhetinic acid reduces renal tubular injury and interstitial inflammatory changes caused by hyperuricemia.

The glycyrrhetinic acid reduces neutrophil infiltration of hyperuricemic renal tissue.

The glycyrrhetinic acid reduces hyperuricemic renal tissue macrophage infiltration.

The glycyrrhetinic acid reduces renal tubular epithelial cell death induced by sodium urate crystals.

The glycyrrhetinic acid reduces the activation of macrophage NLRP3 inflammasome induced by sodium urate crystal, the shearing of cell apoptosis marker protein GSDMD and the secretion of IL-1 beta.

The inventor finds that the glycyrrhetinic acid obviously reduces the renal tubular interstitial inflammation caused by the hyperuricemia by inhibiting the renal tubular epithelial cell necrosis caused by the hyperuricemia, inhibiting the activation of mononuclear macrophage Nlrp3 inflammasome caused by the hyperuricemia crystal, and GSDMD-mediated cell apoptosis and the secretion of inflammatory factors.

Use of a composition comprising glycyrrhetinic acid for the manufacture of a medicament for the treatment of tubulointerstitial inflammation, which may be acute calcium oxalate nephropathy or hyperuricemic nephropathy.

The composition is a medicament prepared from glycyrrhetinic acid as an active ingredient and pharmaceutically acceptable auxiliary materials. .

The glycyrrhetinic acid can be used alone, can be matched with other medicines for use simultaneously, or can be prepared into a compound preparation together with other medicines for use when being used for treating the tubulointerstitial inflammation, and the aim of treating the tubulointerstitial inflammation can be fulfilled.

The pharmaceutically acceptable auxiliary materials refer to various conventional auxiliary materials required when different dosage forms are prepared, such as diluents, adhesives, disintegrants, glidants, lubricants, flavoring agents, inclusion materials, adsorbing materials and the like, and the pharmaceutically acceptable auxiliary materials are prepared into any one of common oral preparations by a conventional preparation method, such as granules, powder, tablets, capsules, pills, oral liquid, decoction, dropping pills and the like.

The pharmacological experiment result shows that:

(1) glycyrrhetinic acid can reduce renal function damage of hyperuricemia nephropathy, and Blood Urea Nitrogen (BUN) and creatinine (Cr) of Glycyrrhetinic acid intervention group are both reduced obviously.

(2) The glycyrrhetinic acid can relieve renal tubular injury and interstitial inflammation change caused by hyperuricemia, and the glycyrrhetinic acid intervention group has the advantages of relieving renal tubular death degree, reducing renal tubular dilation and obviously reducing interstitial inflammatory cell infiltration.

(3) Glycyrrhetinic acid can reduce neutrophil infiltration of hyperuricemic renal tissue, and the number of neutrophils in the glycyrrhetinic acid stem-graft group is obviously reduced. Namely, the number of Ly6G positive cells was significantly reduced.

(4) The glycyrrhetinic acid can reduce macrophage infiltration of hyperuricemia renal tissue, and the number of macrophages in a glycyrrhetinic acid intervention group is obviously reduced. That is, the number of F4/80 positive cells was significantly reduced.

(5) Glycyrrhetinic acid reduces renal tubular epithelial cell death induced by sodium urate crystal, i.e. PI positive (red fluorescence) cell number is obviously increased.

(6) Glycyrrhetinic acid reduces sodium urate crystal-induced activation of macrophage NLRP3 inflammasome, shearing of cell apoptosis marker protein GSDMD and secretion of IL-1 beta.

Drawings

FIG. 1 is a graph showing the effect of glycyrrhetinic acid on renal function of hyperuricemic nephropathy; the left graph in the figure is a BUN graph; the right panel is the Scr map.

FIG. 2 is a graph showing the effect of glycyrrhetinic acid on tubulointerstitial injury in hyperuricemic nephropathy; the PAS staining patterns of the control group are respectively from left to right in the figure; model group PAS staining pattern; a dyeing pattern of PAS in glycyrrhetinic acid group; scoring graph of tubulointerstitial injury.

FIG. 3 is a graph showing the effect of glycyrrhetinic acid on neutrophil infiltration in hyperuricemic nephropathy; from left to right in the figure are immunohistochemical staining patterns of a control group respectively; a model group immunohistochemical staining pattern; immunohistochemical staining pattern of glycyrrhetinic acid group; ly6G positive cell number map.

FIG. 4 is a graph showing the effect of glycyrrhetinic acid on macrophage infiltration of renal tissue of hyperuricemic nephropathy; from left to right in the figure are immunohistochemical staining patterns of a control group respectively; a model group immunohistochemical staining pattern; immunohistochemical staining pattern of glycyrrhetinic acid group; f4/80 positive cell number plot.

FIG. 5 is a graph of the effect of glycyrrhetinic acid on sodium urate-induced renal tubular epithelial cell death; the control group PI/DAPI double staining images are respectively observed from left to right through a fluorescence microscope; model group PI/DAPI double staining pattern; a glycyrrhetinic acid group PI/DAPI double staining pattern; number of PI positive cells. (PI (red): indicating cell death, DAPI (blue): indicating nucleus; Propidium Iodide (PI), DAPI, i.e., 4', 6-diamidino-2-phenylindole, indicating living cells)

FIG. 6 is a graph of glycyrrhetinic acid inhibiting sodium urate-induced activation of macrophage inflammasome and IL-1 β secretion; in the figure, 1 is a control group, 2 is a sodium urate stimulation group, and 3 is a glycyrrhetinic acid + sodium urate stimulation group. The left picture is Western blot protein blot; the right panel shows the secretion level of IL-1. beta. by ELISA.

Detailed Description

The present application will now be described more fully hereinafter with reference to the accompanying drawings, in which some embodiments of the invention are shown and described, but which are not intended to limit the invention thereto.

Pharmacological experimental data of glycyrrhetinic acid for treating tubulointerstitial inflammation

First, test method

(1) Establishment of adenine diet-induced hyperuricemic renal injury: mice were fed a diet containing 0.2% adenine (TD 150071). Mice blood and kidney tissue were taken 21 days later. (this model leads to hyperuricemia, production of uric acid crystals in renal tissue, and tubular interstitial injury)

(2) Glycyrrhetinic acid pretreatment: the administration is carried out 24 hours before the adenine diet, the administration is carried out once per day by intragastric administration of glycyrrhetinic acid 40mg/kg, and the administration is changed to once every other day after 1 week until the adenine diet observation time is over.

(3) The control group was gavaged with physiological saline only and was fed with a conventional diet.

Secondly, the result is:

(1)

BUN (mg/dl): control group (7.896 ± 0.836), model group (66.238 ± 7.186); glycyrrhetinic acid group (30.41 + -6.398)

Scr (mg/dl), control group (0.196 + -0.02), model group (0.94 + -0.142); glycyrrhetinic acid group (0.41 + -0.074)

Glycyrrhetinic acid reduced renal dysfunction of hyperuricemic nephropathy, and both Blood Urea Nitrogen (BUN) and creatinine (Cr) were significantly reduced in Glycyrrhetinic acid intervention group (see FIG. 1).

(2) Glycyrrhetinic acid reduces renal tubular injury and interstitial inflammation change caused by hyperuricemia, and glycyrrhetinic acid intervention group reduces renal tubular death degree, renal tubular dilation and interstitial inflammatory cell infiltration obviously (see figure 2).

(3) Glycyrrhetinic acid can reduce neutrophil infiltration of hyperuricemic renal tissue, and the number of neutrophils in the glycyrrhetinic acid stem-graft group is obviously reduced. I.e., the number of Ly6G positive cells was significantly reduced (see FIG. 3).

(4) The glycyrrhetinic acid can reduce macrophage infiltration of hyperuricemia renal tissue, and the number of macrophages in a glycyrrhetinic acid intervention group is obviously reduced. That is, the number of F4/80 positive cells was significantly reduced (see FIG. 4).

(5) Glycyrrhetinic acid reduced sodium urate crystal-induced renal tubular epithelial cell death, i.e., a significant increase in the number of PI-positive (red fluorescence) cells (see fig. 5).

(6) Glycyrrhetinic acid reduced sodium urate crystal-induced activation of macrophage NLRP3 inflammasome, cleavage of the cell apoptosis marker protein GSDMD and secretion of IL-1 β (see FIG. 6).