A traditional Chinese medicine composition for treating sepsis-related acute kidney injury and a preparation method thereof

Abstract

This invention belongs to the field of traditional Chinese medicine treatment of acute kidney injury, and provides a traditional Chinese medicine composition for treating sepsis-related acute kidney injury and its preparation method. It is prepared by mixing honeysuckle, forsythia, roasted peach kernel, raw rhubarb, raw astragalus, wine-processed angelica, cinnamon twig, mirabilite, and raw licorice in a certain proportion. It detoxifies and removes blood stasis, invigorates qi and unblocks the meridians. It is used for persistent high fever and difficulty in urination and defecation caused by internal accumulation of toxic heat and blood stasis. Symptoms include oliguria or dribbling, red tongue, and rapid pulse; it is also used for sepsis patients with the above symptoms. Treatment with the detoxifying and blood-stasis-removing granules improved survival rate, effectively restored some renal function, reduced renal tubular damage, and decreased TNF-α and IL-1β expression. Immunohistochemical and Western blot results were consistent. The detoxifying and blood-stasis-removing granules can alleviate sepsis-induced acute kidney injury.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine treatment of acute kidney injury, specifically relating to a traditional Chinese medicine composition for treating sepsis-related acute kidney injury and its preparation method. Background Technology

[0002] Sepsis is a life-threatening syndrome of organ dysfunction caused by a dysregulation of the host immune response to infection. Acute kidney injury (AKI) occurring within 7 days of the onset of sepsis is called sepsis-associated acute kidney injury (SA-AKI), characterized by a sudden drop in renal function, elevated creatinine and blood urea nitrogen, and decreased glomerular filtration rate. Despite significant advancements in anti-infective therapy and organ function support in modern medicine, specific treatments for SA-AKI remain scarce. Recent studies have revealed that the pathological process of SA-AKI extends far beyond the traditional understanding of inflammatory storms and microcirculatory disturbances. In critically ill patients, SA-AKI remains a common and fatal complication. Although our understanding of its pathophysiological mechanisms has progressed, timely and effective treatment strategies are still lacking. Recent research evidence suggests that inflammatory response, metabolic reprogramming, and microvascular dysfunction are three potential cornerstones of the pathophysiological mechanism of SA-AKI. Nan et al. conducted immune infiltration analysis on multiple SA-AKI populations, and the results showed significant infiltration of DCS, NK cells, mast cells, and monocytes in the SA-AKI group. In severe sepsis, mast cells far from the initial infection site are also activated, releasing IL-6 and TNF, which exacerbate the inflammatory response and increase sepsis mortality. This indicates that inflammation plays an indispensable role in the development of SA-AKI. Summary of the Invention

[0003] This invention provides a traditional Chinese medicine composition for treating sepsis-related acute kidney injury and its preparation method.

[0004] This invention is achieved by the following technical solution: a traditional Chinese medicine composition for treating sepsis-related acute kidney injury, prepared from the following raw materials in parts by weight: honeysuckle 320-370 parts, forsythia 320-370 parts, roasted peach kernel 200-230 parts, raw rhubarb 260-300 parts, raw astragalus 400-440 parts, wine-processed angelica 200-220 parts, cinnamon twig 120-150 parts, mirabilite 130-150 parts, and raw licorice 130-150 parts.

[0005] Furthermore, it is prepared from the following raw materials in parts by weight: 350 parts honeysuckle, 350 parts forsythia, 210 parts roasted peach kernel, 280 parts raw rhubarb, 420 parts raw astragalus, 210 parts wine-processed angelica, 140 parts cinnamon twig, 140 parts mirabilite, and 140 parts raw licorice.

[0006] The dosage form of the traditional Chinese medicine composition is decoction, tablet or granule.

[0007] Furthermore, the dosage form is granules, and the specific preparation method is as follows: extract rhubarb alone, crush it into coarse granules, soak it in water, decoct for 20 minutes, and obtain a decoction of 0.0776g of Chinese herbal medicine slices per 1ml, with a standard relative density of 1.0083. Filter it, concentrate it under reduced pressure to a preliminary concentrated solution of 1g of Chinese herbal medicine slices per 1ml, with a standard relative density of 1.0870, and set it aside for later use. Honeysuckle, forsythia, roasted peach kernel, raw astragalus, wine-processed angelica, cinnamon twig, and licorice were decocted three times with water. The first decoction was performed simultaneously with distillation, collecting 1.0 ml of volatile oil. 6 g of β-cyclodextrin inclusion complex was prepared by adding the excipient β-cyclodextrin for later use. The first decoction yielded a solution equivalent to 0.1645 g of the medicinal herbs per ml, with a standard relative density of 1.0141. The second decoction yielded a solution equivalent to 0.1645 g of the medicinal herbs per ml, with a standard relative density of 1.0047. The third decoction yielded a solution equivalent to 0.2660 g of the medicinal herbs per ml, with a standard relative density of 1.0021. The decoctions were filtered. The extracts were combined and concentrated under reduced pressure to a primary concentrate equivalent to 1g of Chinese herbal medicine slices per 1ml, with a standard relative density of approximately 1.1250. The resulting primary concentrate was combined with the primary concentrate of rhubarb, dissolved in Glauber's salt, and then concentrated under reduced pressure to a thick paste equivalent to 1.5g of Chinese herbal medicine slices per 1g. The paste was then vacuum-dried at low temperature and pulverized into 696g of crude extract powder. 696g of crude extract powder was taken and 6g of β-cyclodextrin inclusion complex obtained simultaneously during the first distillation was added. The mixture was then pulverized, mixed evenly, and dry-processed into 1000g of granules, each 1g of which was equivalent to 2.240g of Chinese herbal medicine slices, containing 0.696g of the material standard and 6mg of the volatile oil inclusion complex.

[0008] The present invention also provides the application of the traditional Chinese medicine composition for treating SA-AKI in the preparation of a drug that reduces the release of TNF-α and IL-1β inflammatory factors and improves SA-AKI.

[0009] The traditional Chinese medicine composition preparation provided by this invention is named Jiedu Xieyu Keli (JDXYKL), which is derived from the classic formulas of Walnut Decoction for Relieving Stasis and Simiao Decoction. In the early clinical application, it has gradually formed an empirical formula for treating SA-AKI that integrates clearing heat, detoxifying, replenishing qi and nourishing yin, and has achieved good therapeutic effects in clinical practice.

[0010] The traditional Chinese medicine composition of this invention detoxifies, removes blood stasis, invigorates qi, and unblocks the meridians. It is used for persistent high fever and difficulty in urination and defecation caused by internal accumulation of toxic heat and blood stasis. Symptoms include oliguria or dribbling urine, a red tongue, and a rapid pulse; it is also used for sepsis with the above symptoms.

[0011] Dosage and administration: Decocted in water, one dose per day. Decoct twice according to the method, each time yielding 400ml of liquid, to be taken in two divided doses.

[0012] The traditional Chinese medicine composition of this invention is formulated to detoxify, remove blood stasis, invigorate qi, and unblock the meridians. Honeysuckle combined with forsythia serves as the principal herb, clearing heat and relieving exterior symptoms, detoxifying, unblocking qi and blood, and reducing swelling and dissipating nodules. Peach kernel combined with rhubarb invigorates blood, removes blood stasis, and clears heat from the bowels, serving as the assistant herb. Astragalus combined with angelica tonifies the qi of the spleen and lungs, nourishes the source of blood production, and replenishes the blood of the heart and liver, thus tonifying blood and harmonizing the body. Cinnamon twig combined with licorice warms and invigorates the heart yang, tonifies the heart and lungs, and is gentle without being harsh or drying, and nourishing without causing stagnation. Glauber's salt purges and promotes bowel movements, moistens dryness and softens hardness, and assists rhubarb in clearing heat from the bowels, serving as the adjuvant herb. Licorice harmonizes all the herbs and also acts as the guiding herb. The combined herbs work together to detoxify, remove blood stasis, invigorate qi, and unblock the meridians. Attached Figure Description

[0013] Figure 1 The figures show the comparison of survival rate, serum creatinine (Scr), and serum urea nitrogen (BUN) levels among the rat groups. A represents the comparison of survival rate; B represents the comparison of Scr levels; and C represents the comparison of BUN levels. P<0.05, P<0.01, P<0.001; Figure 2 This image shows a comparison of renal tubular injury in different groups of rats. In the figure: A shows the renal tubular staining and injury status in each group; B shows the renal tubular injury score; C shows the NGAL expression electrophoresis image; D shows the statistical results of NGAL expression. P<0.05, P<0.01, P<0.001; Figure 3 The results show the comparison of inflammatory factors in rats from different groups; in the figure: A is the serum TNF-α level measured by ELISA; B is the Western blot result; C is the IL-1β detection result; D is the TNF-α detection result. P<0.05, P<0.01, P<0.001; Figure 4 This figure compares the ATP and lactate levels of rats in each group; in the figure: A represents the ATP detection result; B represents the lactate detection result. P<0.05, P<0.01, P<0.001. Detailed Implementation

[0014] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are some embodiments of the present invention, but not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0015] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains, and all materials cited herein and referenced by them are incorporated herein by reference.

[0016] Equivalent technologies of the specific embodiments described herein that are readily apparent to those skilled in the art through routine experimentation are included in this application.

[0017] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the instruments and equipment used in the following examples are all standard laboratory instruments and equipment; unless otherwise specified, the experimental materials used in the following examples were all purchased from regular biochemical reagent stores.

[0018] Example 1: A detoxifying and blood-stasis-removing decoction for treating SA-AKI, prepared from the following ingredients by weight: honeysuckle 320g, forsythia 320g, raw rhubarb 260g, roasted peach kernel 200g, raw astragalus 400g, wine-processed angelica 200g, cinnamon twig 120g, raw licorice root 130g, and mirabilite 130g. Decoct in water and take one dose daily. Decoct twice according to the method, obtaining 400ml of liquid from each decoction, to be taken in two divided doses.

[0019] Example 2: A detoxifying and blood-stasis-removing granule for treating SA-AKI, prepared from the following raw materials by weight: honeysuckle 350g, forsythia 350g, raw rhubarb 280g, roasted peach kernel 210g, raw astragalus 420g, wine-processed angelica 210g, cinnamon twig 140g, raw licorice root 140g, and mirabilite 140g, totaling 2240g. Decoct in water and take one dose daily. Decoct twice according to the method, obtaining 400ml of liquid from each decoction, to be taken in two divided doses.

[0020] The method for preparing this prescription into granules is as follows: Rhubarb is extracted, crushed into coarse granules, soaked in an appropriate amount of water, and decocted for 20 minutes. Each 1 ml of the decoction yields a concentration equivalent to 0.0776 g of the prepared herbal pieces, with a standard relative density of 1.0083. The decoction is filtered and concentrated under reduced pressure to a preliminary concentrate equivalent to 1 ml of the prepared herbal pieces, with a standard relative density of 1.0870. The remaining seven herbs, including honeysuckle, are decocted three times with water. The first decoction is carried out simultaneously with distillation, collecting 1.0 ml of volatile oil and preparing 6 g of β-cyclodextrin inclusion complex. Prepare the following: Decoction of rhubarb, each 1 ml containing 0.1645 g of medicinal herbs, with a standard relative density of 1.0141; second decoction, each 1 ml containing 0.1645 g of medicinal herbs, with a standard relative density of 1.0047; third decoction, each 1 ml containing 0.2660 g of medicinal herbs, with a standard relative density of 1.0021. Filter the decoctions, combine them, and concentrate under reduced pressure to a preliminary concentrate containing 1 ml of medicinal herbs, with a standard relative density of approximately 1.1250. Combine this concentrate with the above-mentioned rhubarb preliminary concentrate, add 140 g of Glauber's salt to dissolve it, and then concentrate under reduced pressure to a thick paste containing 1.5 g of medicinal herbs per gram. Vacuum dry at low temperature, and pulverize into approximately 696 g of coarse extract powder. Take 696 g of coarse extract powder, add 6 g of the above-mentioned β-cyclodextrin inclusion complex and appropriate excipients, mix and pulverize, mix evenly, and dry-process to form 1000 g of granules. Each gram is equivalent to 2.240g of prepared Chinese medicine slices, containing 0.696g of material standard and 6mg of volatile oil inclusion complex.

[0021] Example 3: A detoxifying and blood-stasis-removing decoction for treating SA-AKI, prepared from the following ingredients by weight: honeysuckle 370g, forsythia 370g, raw rhubarb 300g, roasted peach kernel 230g, raw astragalus 440g, wine-processed angelica 220g, cinnamon twig 150g, raw licorice root 150g, and mirabilite 150g. Decoct in water and take one dose daily. Decoct twice according to the method, obtaining 400ml of liquid from each decoction, to be taken in two divided doses.

[0022] Clinical trials: The clinical control group received basic Western medicine treatment, which included routine basic treatments such as fluid resuscitation, volume management, precise use of antibiotics, vasoactive drugs, mechanical ventilation, diuretics, and hemofiltration, in accordance with the 2021 International Guidelines for the Management of Sepsis and Septic Shock and the 2012 KDOGO Clinical Practice Guidelines for Acute Kidney Injury. At the same time, it was important to protect the function of other organs, such as correcting heart failure and liver failure.

[0023] In addition to the standard Western medicine basic treatment plan, the treatment group also received the Detoxifying and Stasis-Clearing Decoction described in Example 1 of this invention, administered orally or via nasogastric tube, one dose per day for 7 days.

[0024] Diagnostic basis: (1) Meets the diagnostic criteria for sepsis 3.0 (presence of an infection focus + SOFA score ≥2); (2) Meets the KDIGO diagnostic criteria for acute kidney injury stage 1 or 2: Stage 1: SCr increases to 1.5-1.9 times the baseline value within 7 days; or increases by ≥26.5 μmol / L within 48 hours; urine output <0.5 mL (kg·h) for 6-12 hours. Stage 2: SCr increases to 2.0-2.9 times the baseline value within 7 days; urine output <0.5 mL (kg·h) for ≥12 hours.

[0025] (3) It conforms to the syndrome of internal obstruction of blood stasis and toxin in the "Guiding Principles for Clinical Research of New Chinese Medicines". The main symptoms are persistent high fever, decreased urine output, irritability and delirium, and secondary symptoms are sweating, thirst, dry mouth, dark purple tongue with ecchymosis, and deep and slow or deep and wiry pulse.

[0026] Safety assessment: Closely monitor the patient's vital signs and liver and kidney function during treatment. Discontinue use immediately if any abnormal changes in safety indicators or other serious adverse reactions occur, and administer appropriate treatment according to the severity.

[0027] Methods for evaluating therapeutic efficacy: Significant effect: After treatment, clinical symptoms and manifestations of blood stasis and internal obstruction syndrome completely disappeared, urine output recovered significantly, and Scr was less than 1.5 times the baseline. Effective: Clinical symptoms and manifestations of blood stasis and internal obstruction syndrome were significantly reduced, urine output recovered, and Scr exceeded 1.5 times the baseline but was lower than the highest value; Ineffective: The clinical symptoms of blood stasis and internal obstruction syndrome disappeared, but the daily urine output and serum creatinine (Scr) did not decrease significantly.

[0028] Both groups received specific basic Western medicine treatment plans based on their conditions, such as mechanical ventilation, use of vasoactive drugs like norepinephrine and metaraminol, use of antibiotics like meropenem and cefoperazone, use of diuretics like furosemide and spironolactone, and blood purification. The treatment group received an additional 7 days of treatment with a detoxifying and blood-stasis-removing decoction in addition to the basic Western medicine treatment plan. The specific medications used in the basic Western medicine treatment plans for both groups and the corresponding treatment effects are shown in Table 1.

[0029] Table 1: Specific medications and therapeutic effects in the two groups Table 2 shows the comparison of baseline data between the two groups of patients. There were no significant differences between the two groups in terms of gender, age, infection site, and underlying diseases, and the baseline data of the two groups were balanced.

[0030] Table 2. Comparison of baseline data between the two groups of patients. Comparing the main renal chromatogram (SCr) before and after 7 days of treatment in both groups revealed that SCr decreased in both groups after treatment compared to before treatment, with a more significant decrease in the treatment group compared to the control group, suggesting that the Detoxifying and Stasis-Clearing Decoction has a better protective effect on renal function. Urine output in both groups recovered somewhat after treatment. Specific results are shown in Table 3.

[0031] Table 3 Comparison of renal function indicators between the two groups Note: # indicates a P<0.05 difference compared to the control group at the same time point, and # indicates a P<0.05 difference compared to the pre-treatment level in this group. Comparing the inflammatory markers of the two groups before and after 7 days of treatment, it was found that WBC and PCT decreased in both groups after treatment, but the reduction was more significant in the treatment group, and the inflammatory markers tended to return to normal levels after treatment. The specific results are shown in Table 4.

[0032] Table 4 Comparison of inflammatory markers between the two groups Note: # indicates a P<0.05 difference compared to the control group at the same time point, and # indicates a P<0.05 difference compared to the pre-treatment level in this group. Comparing serum and urinary biomarker levels before and after 7 days of treatment in both groups, the anti-inflammatory factor IL-10 significantly increased after treatment, with a more pronounced increase in the treatment group, suggesting that the Detoxifying and Stasis-Clearing Decoction was more effective in relieving inflammation. The early renal tubular injury marker NGAL decreased more significantly in the treatment group after treatment, indicating that the Detoxifying and Stasis-Clearing Decoction was more effective in reducing renal tubular epithelial cell damage in SAKI patients. The results are shown in Table 5.

[0033] Table 5 Comparison of the two groups of biomarkers Note: # indicates a P<0.05 difference compared to the control group at the same time point, and # indicates a P<0.05 difference compared to the pre-treatment level in this group. Animal experiments verify the effectiveness of the detoxifying and blood-stasis-removing granules. I. Experimental Methods 1. Reagents and Drugs: Lipopolysaccharide 055:b5 (L2880, Sigma, USA), Ulinastatin (Guangdong Tianpu Biochemical Co., Ltd., Guangdong, China). The specific formulation of the Detoxifying and Stasis-Clearing Granules JDXYKL is as described in Example 2, prepared by Shanxi Yellow River Company for animal experiments. Creatinine reagent kit (AKNM008M, Boxbio, Beijing, China), Blood Urea Nitrogen Reagent Kit (AKNM002M, Boxbio, Beijing, China), Lactate Reagent Kit (A019-2, Nanjing Jiancheng, Nanjing, China). Enhanced ATP Assay Kit (S0027, Beyotime Biotechnology Co., Ltd., Shanghai, China). Specific antibodies used are shown in Table 6.

[0034] Table 6: Antibody Usage 2. Construction and treatment of SA-AKI model induced by intraperitoneal injection of lipopolysaccharide: 6-8 week old SPF-grade male SD rats weighing 200±20g were purchased from the Fifth Clinical Medical College of Shanxi Medical University (Shanxi, China). All animal experiments in this invention have been approved by the experimental animal ethics committee of the Fifth Clinical Medical College of Shanxi Medical University, with ethics number [No. 2024] (393). Rats were randomly assigned to five groups (n=15), including a control group, an AKI group, a high-dose detoxification and blood-stasis granule treatment group after AKI (AKI+JDXYKL-H), a medium-dose detoxification and blood-stasis granule treatment group after AKI (AKI+JDXYKL-M), and a ulinastatin treatment group after AKI (AKI+UTI). The SA-AKI model was constructed by intraperitoneal injection of lipopolysaccharide (LPS) (10mg / kg). SD rats, each weighing 200g, were administered the following doses of traditional Chinese medicine: Medium dose: 100ml / 60kg × 0.2kg × 6 times = 2ml; High dose: 100ml / 60kg × 0.2kg × 9 times = 3ml. The traditional Chinese medicine treatment groups received the medium and high doses via gavage for 7 consecutive days, while the other groups served as controls via oral water. The positive control group received intraperitoneal injection of ulinastatin at 2000 IU / kg, administered 30 minutes and 4 hours after LPS injection.

[0035] 3. Monitor the general condition and survival rate of rats: The general condition of rats was monitored by observing activity level, feeding behavior, prickling of hair, and eye discharge. Survival rate was determined by calculating the percentage of surviving rats in each group 16 hours after intraperitoneal injection of LPS.

[0036] 4. Determination of blood urea nitrogen, serum creatinine, and lactate levels: Blood was collected from the heart and stored at 4°C for 2 hours to ensure clotting. The samples were then centrifuged at 3000 rpm for 15 minutes at 4°C, and the supernatant serum was collected. Serum creatinine (Scr), blood urea nitrogen (BUN), and lactate levels were measured using the microplate method.

[0037] 5. HE staining of kidney tissue: Fresh kidney tissue was washed with PBS, fixed with 4% paraformaldehyde, then dehydrated with graded ethanol, and finally embedded, sectioned, and the pathological changes of the kidney tissue were observed and scored under a light microscope.

[0038] 6. Immunohistochemistry: TOM20 (1:1000) antibody was applied to tissue sections dewaxed in xylene and embedded in paraffin. Immunoreactivity was observed under an electron microscope.

[0039] 7. ATP content detection: Following the instructions of the enhanced ATP detection kit, a 20ug sample of kidney tissue was homogenized and the Bca protein concentration was measured first. Finally, the luminescence value was detected using an ELISA reader. The results were calculated and statistically analyzed.

[0040] 8. Western blotting: 20 μg kidney tissue samples were taken from each group of rats. The kidney tissue was lysed using a broad-spectrum phosphatase inhibitor (AR1183, BOSTER, Wuhan, China), a protease inhibitor PMSF (AR1192, BOSTER, Wuhan, China), and enhanced RIPA lysis buffer (AR0102-100, BOSTER, Wuhan, China). After processing, the tissue protein was extracted by ultrasonic disruption. Protein concentration was determined using the dicaprin assay (HBCA-500, HYCEMBIO). A uniform loading volume of 30 μg was used for all protein groups. SDS-PAGE electrophoresis was then performed, with 15 μL loaded per well onto 7.5% and 10% sodium dodecyl sulfate-polyacrylamide gels. Electrophoresis was performed at 80V for 1 hour, then at 120V for 30 minutes, followed by transfer at 250mA for 115 minutes. The membrane was then blocked with 5% milk for 1.5 hours. After blocking, it was incubated overnight with primary antibodies NGAL, IL-1β, and TNF-1α. The following day, after three 10-minute washes, the membrane was incubated with secondary antibodies and then developed using an imaging system. The control was β-actin (AC050-20 μL, ABclonal, Wuhan, China).

[0041] 9. Statistical Analysis: All data were analyzed using SPSS 27.0. Fisher's exact test was used for qualitative data with small sample sizes. Quantitative data conforming to a normal distribution were expressed as mean ± standard deviation (x ± s). After ensuring homogeneity, one-way ANOVA was used to assess differences between groups. For quantitative data not conforming to a normal distribution, the Kruskal-Wallis test was used to compare data between groups. A p-value < 0.05 indicated a significant difference. Graphs were generated using GraphPad Prism 9.0.0.

[0042] II. Experimental Results 1. Effects of JDYKL on mortality and renal function in SA-AKI rats: Compared with the control group, the AKI group showed significant reduction in activity and decreased responsiveness, and the survival rate of these rats was significantly lower than that of the control group. Figure 1 A), and Scr was observed in the AKI group. Figure 1 B) and BUN levels ( Figure 1 C) significantly increased. Compared with the LPS group, and compared with the AKI group, JDXYKL treatment reduced the mortality rate of AKI rats and restored the levels of Scr and BUN in the rats. Figure 1AC). This suggests that JDXYKL may alleviate LPS-induced renal function decline.

[0043] 2. Effects of JDXYKL on renal tubular injury in SA-AKI rats: HE staining showed that, compared with the control group, the AKI group exhibited severe renal tubular injury, manifested as swelling of renal tubular epithelial cells, narrowing of the renal tubular lumen and destruction of tubular structure, inflammatory cell infiltration, and cast formation, etc. Figure 2 A), and showed a significantly increased renal tubular injury score ( Figure 2 B). Compared with AKI rats, JDXYKL rats showed some improvement in renal tubular swelling, luminal narrowing, tubular structural damage, and inflammatory cell infiltration in kidney tissue. Furthermore, NGAL is a marker of renal tubular injury and a sensitive early marker of kidney damage. Compared with the control group, its expression was significantly upregulated in the AKI group ( Figure 2 C and D). NGAL expression was significantly downregulated after JDXYKL treatment ( Figure 2 (C and D).

[0044] 3. Effects of JDXYKL on inflammatory factors in SA-AKI rats: Serum TNF-α levels were measured by ELISA. Compared with the control group, the AKI group showed elevated TNF-α levels ( Figure 3 A). Medium doses of JDXYKL significantly reduced TNF-α levels ( Figure 3 A). Western blot results showed that, compared with the control group, the expression of IL-1β and TNF-α in the kidney tissue of AKI rats was increased ( Figure 3 BD). In contrast, the elevation of these inflammatory factors was significantly reduced after JDXYKL treatment ( Figure 3 BD).

[0045] 4. Effects of JDXYKL on energy metabolism in SA-AKI rats: Energy metabolism assays revealed that LPS attack severely depleted the ATP content in kidney tissue, and JDXYKL treatment partially restored it. Figure 4 A). When the body is severely deficient in energy production and cannot efficiently produce ATP through aerobic oxidation, glycolysis becomes the main pathway for ATP production. Lactic acid is the final product of glycolysis. Therefore, a kit was used to detect serum lactate levels. Serum lactate levels were generally elevated in the AKI group, but returned to near normal levels after treatment with JDYKL. Figure 4 B).

[0046] Experimental data from this invention show that LPS-induced sepsis leads to a significant increase in mortality, renal dysfunction (elevated Scr and BUN levels), and histopathological damage. Furthermore, elevated levels of inflammatory factors were also observed. Treatment with JDXYKL reduced sepsis mortality, decreased Scr and BUN levels, and alleviated histopathological damage and inhibited the release of inflammatory factors.

[0047] The prevention and treatment of SA-AKI is quite complex. Many patients have already developed severe renal impairment when they seek medical treatment. Therefore, it is urgent to develop more accurate diagnostic markers and effective therapeutic targets to promote early intervention of SA-AKI. Studies by Li et al. have shown that enhanced glycolysis promotes lactic acid production, and elevated lactic acid is closely related to the deterioration of renal function in AKI patients. Therefore, lactic acid can be used as an independent predictor of the prognosis of AKI patients

[14] . This invention found that in the LPS-induced SA-AKI rat model, the ATP content in the kidney tissue was significantly reduced, accompanied by an increase in serum lactic acid levels. The ATP depletion and elevated lactic acid in the LPS group together constituted a metabolic crisis of "insufficient energy production". In addition, ATP depletion directly weakens the ion transport and reabsorption functions of renal tubular epithelial cells, while the accumulation of lactic acid further aggravates intracellular acidosis and microenvironment disorder. However, JDXYKL treatment can simultaneously increase ATP content and reduce lactic acid levels.

[0048] From the perspective of Traditional Chinese Medicine (TCM) theory, the results of this invention provide a modern biological explanation for the treatment principle of "detoxification, blood stasis removal, and qi tonification." First, "detoxification" is manifested in reducing the inflammatory response caused by LPS toxins, a process achieved by inhibiting the release of inflammatory factors. Second, "blood stasis removal" corresponds to improving "microscopic blood stasis" or "metabolic blockage" represented by lactic acid accumulation, thereby promoting the flow of energy and substances. These two aspects work synergistically to protect the kidneys.

[0049] In summary, JDXYKL can alleviate LPS-induced acute kidney injury, and this alleviating effect mainly depends on reducing the release of inflammatory factors. This invention confirms the therapeutic effect of JDXYKL on SA-AKI and analyzes its possible mechanism, thus providing more scientific evidence for the clinical application of JDXYKL in SA-AKI.

[0050] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A traditional Chinese medicine composition for treating sepsis-related acute kidney injury, characterized in that: It is prepared from the following raw materials in parts by weight: honeysuckle 320-370 parts, forsythia 320-370 parts, roasted peach kernel 200-230 parts, raw rhubarb 260-300 parts, raw astragalus 400-440 parts, wine-processed angelica 200-220 parts, cinnamon twig 120-150 parts, mirabilite 130-150 parts, and raw licorice 130-150 parts.

2. The traditional Chinese medicine composition for treating sepsis-related acute kidney injury according to claim 1, characterized in that: It is prepared from the following raw materials in parts by weight: honeysuckle 350 parts, forsythia 350 parts, boiled peach kernel 210 parts, raw rhubarb 280 parts, raw astragalus 420 parts, wine-processed angelica 210 parts, cinnamon twig 140 parts, mirabilite 140 parts, and raw licorice 140 parts.

3. The traditional Chinese medicine composition for treating sepsis-related acute kidney injury according to claim 1 or 2, characterized in that: The dosage form of the traditional Chinese medicine composition is decoction, tablet or granule.

4. The traditional Chinese medicine composition for treating sepsis-related acute kidney injury according to claim 3, characterized in that: The dosage form is granules, and the specific preparation method is as follows: extract rhubarb alone, crush it into coarse granules, soak it in water, decoct for 20 minutes, and obtain a decoction of 0.0776g of Chinese herbal medicine pieces per 1ml, with a standard relative density of 1.0083. Filter it, concentrate it under reduced pressure to a preliminary concentrate of 1g of Chinese herbal medicine pieces per 1ml, with a standard relative density of 1.0870, and set it aside for later use. Honeysuckle, forsythia, roasted peach kernel, raw astragalus, wine-processed angelica, cinnamon twig, and licorice were decocted three times with water. The first decoction was performed simultaneously with distillation, collecting 1.0 ml of volatile oil. 6 g of β-cyclodextrin inclusion complex was prepared by adding the excipient β-cyclodextrin for later use. The first decoction yielded a solution equivalent to 0.1645 g of the medicinal herbs per ml, with a standard relative density of 1.0141. The second decoction yielded a solution equivalent to 0.1645 g of the medicinal herbs per ml, with a standard relative density of 1.0047. The third decoction yielded a solution equivalent to 0.2660 g of the medicinal herbs per ml, with a standard relative density of 1.0021. The decoctions were filtered. The extracts were combined and concentrated under reduced pressure to a primary concentrate equivalent to 1g of Chinese herbal medicine slices per 1ml, with a standard relative density of approximately 1.1250. The resulting primary concentrate was combined with the primary concentrate of rhubarb, dissolved in Glauber's salt, and then concentrated under reduced pressure to a thick paste equivalent to 1.5g of Chinese herbal medicine slices per 1g. The paste was then vacuum-dried at low temperature and pulverized into 696g of crude extract powder. 696g of crude extract powder was taken and 6g of β-cyclodextrin inclusion complex obtained simultaneously during the first distillation was added. The mixture was then pulverized, mixed evenly, and dry-processed into 1000g of granules, each 1g of which was equivalent to 2.240g of Chinese herbal medicine slices, containing 0.696g of the material standard and 6mg of the volatile oil inclusion complex.

5. The use of the traditional Chinese medicine composition for treating sepsis-related acute kidney injury as described in claim 1 or 2 in the preparation of a medicament for reducing the release of TNF-α and IL-1β inflammatory factors and improving sepsis-related acute kidney injury.

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