阅读说明：本技术 猫爪草提取物制备治疗顺铂致急慢性肾损伤药物中的应用 (Application of radix ranunculi ternati extract in preparation of medicine for treating acute and chronic kidney injury caused by cisplatin ) 是由 刘丽荣 郭兵 徐卫卫 于 2021-01-25 设计创作，主要内容包括：本发明公开了猫爪草提取物制备治疗顺铂致急慢性肾损伤药物中的应用,取猫爪草干燥块根,研磨至粗颗粒,用提取溶媒进行提取,所得提取液除去溶剂后得到猫爪草提取物。该猫爪草提取物在制备治疗顺铂导致的急慢性肾损伤药物中的应用。本发明的猫爪草提取物的制备过程简单易行,作为制备治疗顺铂导致的急慢性肾损伤药物的原料,药理作用强、安全无毒,且其单剂提取物较复合剂提取物更能有效地改善肾脏损伤、维持和修复肾功能。本发明的突出进步和显著效果还在于猫爪草是贵州、河南和云南等省常见易得的中草药,成本低廉,为社会提供了一种具有显著治疗顺铂导致的急慢性肾损伤的天然有效的药物。(The invention discloses an application of radix ranunculi ternati extract in preparation of a medicine for treating acute and chronic kidney injury caused by cisplatin. The application of the radix ranunculi ternati extract in preparing medicines for treating acute and chronic kidney injury caused by cisplatin is provided. The preparation process of the radix ranunculi ternati extract is simple and easy to implement, the radix ranunculi ternati extract is used as a raw material for preparing the medicine for treating acute and chronic kidney injury caused by cisplatin, the pharmacological effect is strong, the radix ranunculi ternati extract is safe and non-toxic, and the single-dose extract can effectively improve the kidney injury and maintain and repair the kidney function compared with a compound-dose extract. The invention has the outstanding advantages and obvious effects that the catclaw buttercup root is a common and easily-obtained Chinese herbal medicine in Guizhou, Henan, Yunnan and the like, has low cost and provides a natural and effective medicine for remarkably treating acute and chronic kidney injury caused by cis-platinum for the society.)

1. The radix ranunculi ternati extract is characterized in that: radix Ranunculi Ternati is dry root tuber of small hair food of family Oesophagaceae.

2. The method for preparing the catclaw buttercup root extract according to claim 1, wherein: drying radix Ranunculi Ternati, grinding to coarse particles, extracting with extraction solvent, and removing solvent from the obtained extractive solution to obtain radix Ranunculi Ternati extract.

3. The method for preparing the catclaw buttercup root extract according to claim 2, wherein: the extraction solvent is 70% absolute ethyl alcohol.

4. The method for preparing the catclaw buttercup root extract according to claim 2, wherein: the extraction method comprises heating, refluxing and cold soaking at 80 deg.C.

5. The method for preparing the catclaw buttercup root extract according to claim 2, wherein: the method for removing solvent from extractive solution adopts reduced pressure steam concentration.

6. Application of radix Ranunculi Ternati extract in preparing medicine for treating acute and chronic kidney injury caused by cisplatin is provided.

7. The use of the radix ranunculi ternati extract in the preparation of a medicament for treating acute and chronic kidney injury caused by cisplatin according to claim 6, wherein the application comprises the following steps: the radix ranunculi ternati extract can effectively improve renal function and relieve acute and chronic renal injuries, and the action mechanism of the radix ranunculi ternati extract is probably closely related to the inhibition of 36 th lysine trimethylation (H3K36me3) protein expression of histone H3, signal transduction and transcription regulatory factor (STAT3) and nuclear transcription factor kappa B (NF-kappa B) signal transduction pathway activation, and further the inhibition of renal tubular cell apoptosis and inflammatory reaction.

Technical Field

The invention relates to a natural Chinese herbal medicine extract, in particular to a radix ranunculi ternati extract, a preparation method of the radix ranunculi ternati extract and application of the radix ranunculi ternati in preparation of a medicine for treating acute and chronic kidney injury caused by cisplatin. The Ranunculus ternatus is particularly a dry root tuber of Ranunculus ternatus Thunb belonging to Ranunculaceae.

Background

Cisplatin is one of the most common clinical antitumor drugs, and is widely used for multiple solid tumors such as ovarian cancer, non-small cell lung cancer, bladder cancer, head and neck squamous cell carcinoma and the like. Cisplatin chemotherapy, however, has a number of serious adverse effects, such as nephrotoxicity, ototoxicity, neurotoxicity, nausea and vomiting, among which nephrotoxicity is the most common and most serious toxic effect, with an incidence of up to 30% (nutriments, 2018). Researches show that single large-dose cisplatin stimulation can cause renal tubular epithelial cell apoptosis to cause acute renal injury, while large-dose or continuous multiple small-dose cisplatin stimulation can cause renal tubular necrosis, interstitial injury and the like to cause TGF-beta 1 and other proteins to be expressed in the kidney and increase extracellular matrix to mediate fibroblast to be converted into myofibroblast, and thus chronic renal disease is caused (Archives of toxicity, 2009, 381-387). Therefore, how to reduce the renal toxicity caused by cisplatin, better exert the anti-tumor effect of cisplatin and improve the chemotherapy curative effect and the life quality of patients is a problem which needs to be solved urgently at present.

The pathological mechanisms of cisplatin-induced kidney injury are complex, with tubular injury, oxidative stress and inflammatory responses all being key factors leading to acute kidney injury (Journal of biological science, 2019, 25). A variety of different pathological responses can induce tubular injury, such as: apoptosis, necrosis, inflammation, and activation of signal transduction and transcriptional activator (STAT3) or nuclear transcription factor kappa B (NF-. kappa.B) signaling pathways (Journal of aggregative and food chemistry, 2019, 5754-one 5763). There have been numerous reports on the role of tubular apoptosis in cisplatin-acute kidney injury. Compared with cisplatin-acute kidney injury, the research on cisplatin-chronic kidney injury at home and abroad is relatively less at present, and the specific mechanism is not clear. The research shows that repeated small-dose or large-dose intraperitoneal injection of cisplatin can cause renal dysfunction, renal interstitial fibrosis and renal injury. Renal fibrosis is mainly manifested by excessive cell deposition and epithelial-mesenchymal transdifferentiation, etc., which ultimately leads to renal parenchyma destruction and renal function loss (Ebiomedicine,2019, 491-504). In conclusion, drug-induced kidney injury, especially acute and chronic kidney injury caused by cisplatin, has become the most common clinical problem, and seriously affects the use of clinical chemotherapeutic drugs, the treatment effect of tumors, and the structure and function of kidneys. However, the pathological mechanism is still unclear at present, and the search of effective therapeutic drugs is crucial.

The Ranunculus ternatus (L.) nakai is dry root tuber of Ranunculus ternatus (Ranunculus ternatus Thunb.) of Ranunculus of Ranunculaceae, and is spindle-shaped, has 5-6 clusters, is similar to Ranunculus ternatus L.var, and has length of 3-10mm, diameter of 2-3mm, and yellowish brown residue or stem mark at top. The surface is yellow brown or gray yellow, the color becomes dark after long-term storage, and the hair has slight longitudinal wrinkles, a bit of fibrous root marks and residual fibrous roots. Firm, white or yellowish white cross section, solid, powdery. Slightly sweet in taste. The Sandaliancao, the Uncaria tomentosa and the Uncaria tomentosa are mainly produced in Xinyang in Henan, and are widely distributed in Guizhou and Yunnan provinces. The medicine is warm in nature, sweet and pungent in taste, enters liver and lung channels, has the effects of clearing heat and removing toxicity, reducing swelling and resolving masses, relieving cough and eliminating phlegm and the like, and is clinically used for treating diseases such as pulmonary tuberculosis, tuberculous lymphadenitis, sphagitis, malaria and the like (the university of Guangdong pharmacy, 2019, 1-5). The medicine is used for treating scrofula in folk, and has good curative effect no matter whether tuberculosis is large or suppurative.

There are many descriptions about the use of a single-dose extract or a combined-dose extract of Ranunculi Ternati as a Chinese herbal medicine. The single-dose extract of the radix ranunculi ternati has good effect on treating the scrofula; the single injection has the inhibiting effect on mouse S180 and S37 cancer strains; has obvious inhibiting effect on dysentery bacillus, staphylococcus aureus, staphylococcus albus, tetragenococcus and the like (Chinese hospital pharmacy journal, 2011, 1673-; also has significant anti-tumor effect (a preparation method of the anti-tumor component of the radix ranunculi ternati, CN105412265A, 2018). However, the function and possible mechanism of the single-dose extract of the radix ranunculi ternati in acute and chronic kidney injury are not reported in related researches at present. Researches show that a compound preparation prepared by mixing and decocting various traditional Chinese medicine components such as radix ranunculi ternati, fructus cannabis, semen decanomiae, magnolia sieboldii and the like with water can effectively prevent and treat diabetic nephropathy, but a specific mechanism is not clarified (a preparation method of a diabetic nephropathy medicine, CN104815194A, 2015). Similarly, the mixture obtained by mixing and decocting the traditional Chinese medicine components such as the radix ranunculi ternati, the gecko, the saffron, the centipede, the roasted radix stemonae and the like can effectively treat the renal tuberculosis, the action mechanism of the mixture is related to the regulation of the immunity of the organism, the anti-inflammation and the anti-tuberculosis, the curative effect is obvious, and no toxic or side effect exists (a traditional Chinese medicine preparation for treating the renal tuberculosis, CN1911411 and 2007). The mixture prepared by steaming and boiling the radix ranunculi ternati, the rhizoma paridis, the semen plantaginis and the radix astragali into paste can effectively treat acute and chronic nephritis and urinary tract infection radically, is related to rapid inflammation diminishing, swelling diminishing and toxin expelling, but the specific mechanism is not clear (traditional Chinese medicine preparation for treating nephritis, CN101904970A and 2010). The research and the related application of the single-dose or compound-dose extract of the radix ranunculi ternati for treating the acute and chronic kidney injury of the mice caused by the cisplatin are not reported. Therefore, the invention provides a natural effective medicament capable of effectively treating acute and chronic kidney injury induced by cisplatin, and elucidates the possible action mechanism of the medicament, and the medicament has important effect on the future prevention and treatment of the acute and chronic kidney injury.

Research has found that epigenetics plays an important role in regulating the development of kidney diseases (Nature reviews neuropathology, 2019, 327-. Epigenetics refers to heritable phenotypic changes in genes that occur under various controls such as DNA modification, protein modification, and non-coding RNA without alterations in DNA sequences (worldwide latest medical information abstracts, 2016, 58-59). Histone methylation is one of the studies in epigenetics, and is a reversible enzymatic reaction catalyzed by lysine methyltransferases, and some lysine methyltransferases can catalyze methylation of both histone (e.g., lysine 36 (H3K36) of histone H3 or lysine 4 (H3K4) of histone H3) and non-histone (e.g., signal transducer and activator of transcription (STAT3) or nuclear transcription factor kappa B (NF-kappa B)) to perform their biological functions. Many sites on histones (especially lysine) can be modified by methylation, and methylation at different sites can be catalyzed by different lysine methyltransferases, resulting in inhibition or activation of gene transcription. The expression of lysine trimethylation at position 36 of histone H3 (H3K36me3) is up-regulated in renal epithelial cells of autosomal dominant polycystic kidney patients and mice, and The up-regulation of The expression is closely related to The occurrence of renal cyst formation and renal tubular apoptosis (The Journal of clinical invasion, 2017, 2751-2764). However, the expression change and possible action mechanism of the 36 th lysine trimethylation (H3K36me3) of the histone H3 in the acute and chronic kidney injury induced by the cisplatin are not reported in the research.

In conclusion, cisplatin is an effective anti-tumor chemotherapeutic drug, but has serious nephrotoxicity, so that the clinical application of the cisplatin is limited. At present, the pathogenesis of the cisplatin-induced acute and chronic kidney injury is not completely clarified, and researches show that the cisplatin-induced acute and chronic kidney injury has a significant correlation with epigenetics. In recent years, the therapeutic effect of the radix ranunculi ternati extract in various diseases is increasingly remarkable and receives wide attention, but the effect of the radix ranunculi ternati extract in the induction of acute and chronic kidney injury by cisplatin injection is not reported. Therefore, the invention provides a natural effective medicament for remarkably treating acute and chronic kidney injury caused by cisplatin for the society, and clarifies a possible action mechanism of the medicament.

Disclosure of Invention

The invention aims to solve the technical problems of providing the ternate buttercup root extract, the preparation method thereof and the application thereof in preparing the medicine for treating the acute and chronic kidney injury caused by the cisplatin, providing a new medicine for treating the acute and chronic kidney injury caused by the cisplatin, and having the advantages of easily available raw materials, simple preparation process, high medicinal value and remarkable medicinal effect.

The technical scheme of the invention is as follows: in order to solve the technical problems, the invention provides a radix ranunculi ternati extract and a preparation method of the radix ranunculi ternati extract.

The radix ranunculi ternati extract is prepared by the following process: drying radix Ranunculi Ternati, grinding to coarse particles, extracting with extraction solvent, and removing solvent from the extractive solution to obtain radix Ranunculi Ternati extract.

The extraction solvent is 70% absolute ethyl alcohol.

The extraction method comprises heating, refluxing and cold soaking at 80 deg.C.

The solvent of the extractive solution is removed by steam concentration under reduced pressure.

The invention also provides application of the ternate buttercup root extract in preparing a medicine for treating acute and chronic kidney injury caused by cisplatin.

The radix ranunculi ternati extract is simple and easy to prepare, is used as a raw material for preparing a medicine for treating acute and chronic kidney injury caused by cisplatin, has strong pharmacological action, is safe and non-toxic, and can effectively relieve kidney injury, maintain and repair kidney functions compared with a composite preparation. The mouse model experiment result of acute kidney injury caused by cisplatin shows that the radix ranunculi ternati extract can obviously improve the kidney function of a mouse, down-regulates the protein expression of acute kidney injury marker protein (NGAL), apoptosis protein (Bax and clear caspase-3) and up-regulates the protein expression of anti-apoptosis protein (BCL-2), has obvious anti-apoptosis function, has the action mechanism possibly closely related to the inhibition of the signal transduction and the activation of a signal transduction pathway of a transcription regulatory factor (STAT3), and has the molecular mechanism possibly playing a role in relieving acute kidney injury by inhibiting the expression of 36 th lysine trimethylation (H3K36me3) protein of histone H3. The mouse model experiment result of renal fibrosis caused by cisplatin shows that the catclaw buttercup root extract can obviously reduce the expression of fibrosis protein (Fibronectin and Vimentin), has obvious anti-fibrosis effect, and the action mechanism of the catclaw buttercup root extract is probably closely related to the inhibition of the activation of a nuclear transcription factor kappa B (NF-kappa B) signal transduction path so as to relieve the inflammatory reaction of renal tubular epithelial cells. The remarkable progress and remarkable effect of the invention are that the catclaw buttercup root is a common and easily-obtained Chinese herbal medicine in Henan, Guizhou, Yunnan and the like, has low cost, and provides a natural and effective medicine for remarkably treating acute and chronic kidney injury caused by cisplatin for the society.

Drawings

FIG. 1: BUN and Scr water profiles of groups of C57BL/6 mice;

FIG. 2: each group of C57BL/6 mice acute kidney injury marker protein (NGAL) expression level map;

FIG. 3: graphs of the expression levels of apoptotic protein (Bax and cleaned caspase-3) and anti-apoptotic protein (BCL-2) in groups of C57BL/6 mice;

FIG. 4: graphs of signal transducer and activator of transcription (STAT3) expression levels in kidney tissue of groups C57BL/6 mice;

FIG. 5: the expression level of 36 th lysine trimethylation (H3K36me3) of histone H3 in kidney tissues of each group of C57BL/6 mice is shown;

FIG. 6: graph of renal fibrosis protein (fibrinectin and Vimentin) expression levels in each group of C57BL/6 mice;

FIG. 7: clear caspase-3, Bax and BCL-2 expression level profiles for each group of C57BL/6 mice;

FIG. 8: a graph of tumor necrosis factor (TNF-. alpha.), interleukin-6 (IL-6) and nuclear transcription factor kappa B (NF-. kappa.B) levels in kidney tissue of each group of C57BL/6 mice.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. The preparation of the related pharmacological activity and pharmaceutical preparation of the ternate buttercup root extract of the present invention is performed according to the method shown in the following examples. The methods involved are technical means that can be grasped and utilized by those skilled in the art. However, the following specific examples should not be construed as limiting the claims in any way.

Example 1: preparation of Ranunculus ternatus thumb extract sample

Drying radix Ranunculi Ternati, grinding to coarse particles, extracting with 70% anhydrous ethanol as extraction solvent at 80 deg.C under reflux, cold soaking, and concentrating the extractive solution under reduced pressure to remove solvent to obtain radix Ranunculi Ternati extract. The specific raw materials and their weights, the extraction solvent used, the extraction method used, the solvent removal method, etc. are shown in table 1 below.

TABLE 1 methods of making extracts of Ranunculus ternatus

Example 2: the effect of the ternate buttercup root extract on the kidney function of mice with acute kidney injury caused by cisplatin.

6-8 weeks old C57BL/6 mice were fed adaptively for 1 week, and were randomly divided into a control group (normal + double distilled water), a cisplatin-induced acute kidney injury model group (cisplatin 20mg/kg + double distilled water), and a Ranunculus ternatus thumb treatment group (cisplatin 20mg/kg +2.5 g/kg) with 7 mice per group. A mouse model of acute kidney injury is constructed by a single intraperitoneal injection of cisplatin (20mg/kg) (dissolved in 0.9% physiological saline); mice in the control group were injected with an equal volume of cisplatin vehicle (0.9% physiological saline) intraperitoneally. The ternate buttercup root treatment group is subjected to intragastric gavage by a ternate buttercup root extract (2.5g/kg) 2 hours before cisplatin injection, 1 time/day and 3 continuous days, and the control group and the mice in the cisplatin-induced acute kidney injury model group are subjected to intraperitoneal injection of equal volume double distilled water. Collecting a mouse serum sample in a 3-gastrodia elata drunk state after cisplatin injection, and detecting the content of urea nitrogen (BUN) and creatinine (Scr) in the serum by using a full-automatic biochemical analyzer.

Referring to the BUN and Scr levels in each group of C57BL/6 mice in FIG. 1 (note: vs control group,. times.P)<0.05; a vs cisplatin model group is used as a model group,^#P<0.05; BUN: urea nitrogen; and (2) Scr: (ii) blood creatinine; RTT: radix Ranunculi Ternati extract)

The biochemical results are shown in FIG. 1 (A): the cisplatin-induced acute kidney injury model group mice have obviously increased urea nitrogen (BUN) content and obviously worsened renal function, while the ternate buttercup root extract can obviously reduce urea nitrogen (BUN) level, and the results in figure 1(B) show that: the radix ranunculi ternati extract can obviously reduce the creatinine (Scr) level, which indicates that the radix ranunculi ternati extract has obvious improvement effect on renal function, so that the radix ranunculi ternati extract can obviously reduce the urea nitrogen (BUN) and creatinine (Scr) levels of mice with acute renal injury caused by cisplatin.

Example 3: the ternate buttercup root extract has a protective effect on acute renal injury mouse renal tubular injury caused by cisplatin.

6-8 weeks old C57BL/6 mice were fed adaptively for 1 week, and were randomly divided into a control group (normal + double distilled water), a cisplatin-induced acute kidney injury model group (cisplatin 20mg/kg + double distilled water), and a Ranunculus ternatus thumb treatment group (cisplatin 20mg/kg +2.5 g/kg) with 7 mice per group. A mouse model of acute kidney injury is constructed by a single intraperitoneal injection of cisplatin (20mg/kg) (dissolved in 0.9% physiological saline); mice in the control group were injected with an equal volume of cisplatin vehicle (0.9% physiological saline) intraperitoneally. The ternate buttercup root treatment group is subjected to intragastric gavage by a ternate buttercup root extract (2.5g/kg) 2 hours before cisplatin injection, 1 time/day and 3 continuous days, and the control group and the mice in the cisplatin-induced acute kidney injury model group are subjected to intraperitoneal injection of equal volume double distilled water. Collecting mouse kidney tissue samples under the state of 3 Gastrodia elata drunkenness after cisplatin injection, and extracting total tissue protein.

Referring to the NGAL expression level of each group of C57BL/6 mice in FIG. 2, (note: vs control group,. times.P)<0.05; a vs cisplatin model group is used as a model group,^#P<0.05; RTT: a catclaw buttercup root extract; NGAL: acute kidney injury marker protein

The results are shown in FIG. 2: the cisplatin-induced acute kidney injury model group has the advantages that the acute kidney injury marker protein (NGAL) level is obviously increased, and the ternate buttercup root extract can obviously reduce the acute kidney injury marker protein (NGAL) level, so that the ternate buttercup root is prompted to have the effect of reducing the kidney injury.

Example 4: the ternate buttercup root extract has an inhibiting effect on acute renal injury mouse renal tubular cell apoptosis caused by cisplatin.

6-8 weeks old C57BL/6 mice were fed adaptively for 1 week, and were randomly divided into a control group (normal + double distilled water), a cisplatin-induced acute kidney injury model group (cisplatin 20mg/kg + double distilled water), and a Ranunculus ternatus thumb treatment group (cisplatin 20mg/kg +2.5 g/kg) with 7 mice per group. A mouse model of acute kidney injury is constructed by a single intraperitoneal injection of cisplatin (20mg/kg) (dissolved in 0.9% physiological saline); mice in the control group were injected with an equal volume of cisplatin vehicle (0.9% physiological saline) intraperitoneally. The ternate buttercup root treatment group is subjected to intragastric gavage by a ternate buttercup root extract (2.5g/kg) 2 hours before cisplatin injection, 1 time/day and 3 continuous days, and the control group and the mice in the cisplatin-induced acute kidney injury model group are subjected to intraperitoneal injection of equal volume double distilled water. Collecting mouse kidney tissue samples under the state of 3 Gastrodia elata drunkenness after cisplatin injection, and extracting total tissue protein.

Referring to FIG. 3, the expression levels of apoptotic protein (Bax and cleaned caspase-3) and anti-apoptotic protein (BCL-2) in each group of C57BL/6 mice (note: vs control group, P<0.05; a vs cisplatin model group is used as a model group,^#P<0.05; RTT: a catclaw buttercup root extract; bax, clear caspase-3: an apoptotic protein; BCL-2: anti-apoptotic proteins

The results are shown in FIG. 3: the cisplatin-induced acute kidney injury model group cell apoptosis protein (Bax and cleared caspase-3) expression level is remarkably increased, the anti-apoptosis protein (BCL-2) expression level is remarkably reduced, the ternate buttercup root extract can remarkably reduce the cell apoptosis protein (Bax and cleared caspase-3) expression level, increase the anti-apoptosis protein (BCL-2) expression level and reduce cell apoptosis, and the phenomenon is one of the action mechanisms of the ternate buttercup root extract for protecting acute kidney injury.

Example 5: effect of Uncaria tomentosa extract on Signal transducer and activator of transcription (STAT3) in renal tissue of mice with acute renal injury caused by cisplatin.

6-8 weeks old C57BL/6 mice were fed adaptively for 1 week, and were randomly divided into a control group (normal + double distilled water), a cisplatin-induced acute kidney injury model group (cisplatin 20mg/kg + double distilled water), and a Ranunculus ternatus thumb treatment group (cisplatin 20mg/kg +2.5 g/kg) with 7 mice per group. A mouse model of acute kidney injury is constructed by a single intraperitoneal injection of cisplatin (20mg/kg) (dissolved in 0.9% physiological saline); mice in the control group were injected with an equal volume of cisplatin vehicle (0.9% physiological saline) intraperitoneally. The ternate buttercup root treatment group is subjected to intragastric gavage by a ternate buttercup root extract (2.5g/kg) 2 hours before cisplatin injection, 1 time/day and 3 continuous days, and the control group and the mice in the cisplatin-induced acute kidney injury model group are subjected to intraperitoneal injection of equal volume double distilled water. Collecting mouse kidney tissue samples under the state of 3 Gastrodia elata drunkenness after cisplatin injection, and extracting total tissue protein.

Referring to FIG. 4, the kidney tissue expression levels of signal transducer and activator of transcription (STAT3) in each group of C57BL/6 mice (note: vs control group,. about.P)<0.05; a vs cisplatin model group is used as a model group,^#P<0.05; RTT: a catclaw buttercup root extract; STAT 3: signal transduction and transcriptional activators; p-STAT 3: phosphorylated signal transduction and transcriptional activators

The results are shown in FIG. 4: the levels of a signal transduction and transcriptional activation factor (STAT3) and a phosphorylated signal transduction and transcriptional activation factor (p-STAT3) in a cisplatin model group are remarkably increased, and the ternate buttercup root extract can obviously reduce the levels of the signal transduction and transcriptional activation factor (STAT3) and the phosphorylated signal transduction and transcriptional activation factor (p-STAT3), so that the ternate buttercup root extract is suggested to inhibit the activation of a signal transduction and transcriptional activation factor (p-STAT3) signal transduction pathway, further inhibit the renal tubular cell apoptosis and relieve cisplatin-induced acute kidney injury.

Example 6: effect of Uncaria tomentosa extract on expression of lysine trimethylation at position 36 (H3K36me3) of histone H3 in C57BL/6 mouse cisplatin model tubular epithelial cells.

6-8 weeks old C57BL/6 mice were fed adaptively for 1 week, and were randomly divided into a control group (normal + double distilled water), a cisplatin-induced acute kidney injury model group (cisplatin 20mg/kg + double distilled water), and a Ranunculus ternatus thumb treatment group (cisplatin 20mg/kg +2.5 g/kg) with 7 mice per group. A mouse model of acute kidney injury is constructed by a single intraperitoneal injection of cisplatin (20mg/kg) (dissolved in 0.9% physiological saline); mice in the control group were injected with an equal volume of cisplatin vehicle (0.9% physiological saline) intraperitoneally. The ternate buttercup root treatment group is subjected to intragastric gavage by a ternate buttercup root extract (2.5g/kg) 2 hours before cisplatin injection, 1 time/day and 3 continuous days, and the control group and the mice in the cisplatin-induced acute kidney injury model group are subjected to intraperitoneal injection of equal volume double distilled water. Collecting mouse kidney tissue samples under the state of 3 Gastrodia elata drunkenness after cisplatin injection, and extracting total tissue protein.

Referring to FIG. 5, the expression level of lysine trimethylation at position 36 (H3K36me3) of histone H3 in kidney tissue of each group C57BL/6 mice (note: vs control group,. times.P)<0.05; the group of vs is down-platinum,^#P<0.05; RTT: a catclaw buttercup root extract; H3K36me3 histone H3 lysine trimethylation at position 36 (H3K36me3)

The results are shown in FIG. 5: the level of lysine trimethylation at 36 th position of histone H3 (H3K36me3) in a cisplatin model group is obviously increased, and the ternate buttercup root extract can obviously reduce the level of lysine trimethylation at 36 th position of histone H3 (H3K36me3), so that the ternate buttercup root extract is suggested to relieve acute kidney injury induced by cisplatin from a molecular level by inhibiting the expression of lysine trimethylation at 36 th position of histone H3 (H3K36me 3).

Example 7: the effect of ternate buttercup root extract on the expression of fibrosis-associated proteins in kidney tissue of mice with renal fibrosis caused by cisplatin.

6-8 weeks old C57BL/6 mice were fed adaptively for 1 week, and were randomly divided into a control group (normal + double distilled water), a cisplatin-induced renal fibrosis model group (cisplatin 10mg/kg + double distilled water), and a Ranunculus ternatus thumb treatment group (cisplatin 10mg/kg +1.25 g/kg) with 7 mice per group. Repeatedly injecting cisplatin (10mg/kg) into abdominal cavity (dissolved in 0.9% normal saline) for 3 weeks and 1 time/week to construct a mouse model of chronic renal fibrosis; mice in the control group were injected with an equal volume of cisplatin vehicle (0.9% physiological saline) intraperitoneally. The treatment group of Ranunculus ternatus is administered with Ranunculus ternatus extract (1.25g/kg) by intragastric administration 1 time/day for 21 consecutive days from the day of cisplatin injection. The control group and the cis-platinum induced renal fibrosis model group are subjected to equal double-distilled water intragastric administration. Mouse kidney tissue samples were harvested 21 days after the first cisplatin injection under anesthesia and total tissue protein was extracted.

Referring to the expression levels of renal fibrosis proteins (fibrinectin and Vimentin) in each group of C57BL/6 mice in FIG. 6 (note: vs control group,. times.P)<0.05; a vs cisplatin model group is used as a model group,^#P<0.05; RTT: a catclaw buttercup root extract; fibrinectin, Vimentin: renal fibrosis protein)

The results are shown in FIG. 6: the cisplatin-induced kidney fibrosis model group has the advantages that the expression of the fibrosis proteins (Fibronectin and Vimentin) is remarkably increased, the ternate buttercup root extract can obviously reduce the level of the fibrosis proteins (Fibronectin and Vimentin), and the RTT is suggested to have the function of relieving kidney fibrosis.

Example 8: the ternate buttercup root extract has an inhibiting effect on renal fibrosis mouse renal tubular cell apoptosis caused by cisplatin.

6-8 weeks old C57BL/6 mice were fed adaptively for 1 week, and were randomly divided into a control group (normal + double distilled water), a cisplatin-induced renal fibrosis model group (cisplatin 10mg/kg + double distilled water), and a Ranunculus ternatus thumb treatment group (cisplatin 10mg/kg +1.25 g/kg) with 7 mice per group. Repeatedly injecting cisplatin (10mg/kg) into abdominal cavity (dissolved in 0.9% normal saline) for 3 weeks and 1 time/week to construct a mouse model of chronic renal fibrosis; mice in the control group were injected with an equal volume of cisplatin vehicle (0.9% physiological saline) intraperitoneally. The treatment group of Ranunculus ternatus is administered with Ranunculus ternatus extract (1.25g/kg) by intragastric administration 1 time/day for 21 consecutive days from the day of cisplatin injection. The control group and the cis-platinum induced renal fibrosis model group are subjected to equal double-distilled water intragastric administration. Mouse kidney tissue samples were harvested 21 days after the first cisplatin injection under anesthesia and total tissue protein was extracted.

Referring to FIG. 7, the expression levels of apoptotic proteins (cleaned caspase-3 and Bax) and anti-apoptotic proteins (BCL-2) in each group of C57BL/6 mice (note: vs control group,. times.P)<0.05; a vs cisplatin model group is used as a model group,^#P<0.05; RTT: a catclaw buttercup root extract; clean caspase-3, Bax: an apoptotic protein; BCL-2: anti-apoptotic proteins

The results are shown in FIG. 7: the cisplatin-induced renal fibrosis model group has the advantages that the levels of apoptosis proteins (cleared caspase-3 and Bax) are obviously increased, the level of anti-apoptosis protein (BCL-2) is obviously reduced, the ternate buttercup root extract can obviously reduce the expression levels of the apoptosis proteins (cleared caspase-3 and Bax), increase the expression level of the anti-apoptosis protein (BCL-2) and reduce apoptosis, and the cisplatin-induced renal fibrosis model group is one of the action mechanisms of the ternate buttercup root extract for protecting chronic renal injury.

Example 9: effect of Uncaria tomentosa extract on levels of tumor necrosis factor (TNF- α), interleukin-6 (IL-6), and nuclear transcription factor κ B (NF- κ B) in kidney tissues of mice with renal fibrosis induced by cisplatin.

6-8 weeks old C57BL/6 mice were fed adaptively for 1 week, and were randomly divided into a control group (normal + double distilled water), a cisplatin-induced renal fibrosis model group (cisplatin 10mg/kg + double distilled water), and a Ranunculus ternatus thumb treatment group (cisplatin 10mg/kg +1.25 g/kg) with 7 mice per group. Repeatedly injecting cisplatin (10mg/kg) into abdominal cavity (dissolved in 0.9% normal saline) for 3 weeks and 1 time/week to construct a mouse model of chronic renal fibrosis; mice in the control group were injected with an equal volume of cisplatin vehicle (0.9% physiological saline) intraperitoneally. The treatment group of Ranunculus ternatus is administered with Ranunculus ternatus extract (1.25g/kg) by intragastric administration 1 time/day for 21 consecutive days from the day of cisplatin injection. The control group and the cis-platinum induced renal fibrosis model group are subjected to equal double-distilled water intragastric administration. Mouse kidney tissue samples were harvested 21 days after the first cisplatin injection under anesthesia and total tissue protein was extracted.

Referring to FIG. 8, the levels of tumor necrosis factor (TNF-. alpha.), interleukin-6 (IL-6) and nuclear transcription factor kB (NF-. kappa.B) in kidney tissues of each group of C57BL/6 mice (note: vs control group,. times.P)<0.05; a vs cisplatin model group is used as a model group,^#P<0.05; RTT: a catclaw buttercup root extract; TNF- α, IL-6, NF-. kappa. B p-p65 and NF-. kappa. B p 65: inflammation reaction-related protein

The results are shown in FIG. 8: the cisplatin-induced renal fibrosis model group has the advantages that the levels of inflammatory response related proteins (TNF-alpha, IL-6, NF-kappa B p-p65 and NF-kappa B p65) are remarkably increased, and the ternate buttercup root extract can obviously reduce the expression levels of the inflammatory response related proteins (TNF-alpha, IL-6, NF-kappa B p-p65 and NF-kappa B p65), so that the ternate buttercup root extract can inhibit inflammatory response and relieve cisplatin-induced renal fibrosis by inhibiting the activation of a nuclear transcription factor kappa B (NF-kappa B) signal transduction pathway.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.