A traditional Chinese medicine composition for treating sepsis cardiomyopathy and application thereof

Abstract

The application belongs to the field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine composition for treating sepsis cardiomyopathy and application thereof, which mainly comprises, by mass fraction, 10-50 parts of ginseng, 5-20 parts of rhubarb, 10-30 parts of monkshood, 5-20 parts of coptis, 5-30 parts of scutellaria and 5-20 parts of safflower. Clinical tests show that the traditional Chinese medicine composition can not only improve circulation and heart function of patients with sepsis yang deficiency and blood stasis syndrome, reduce inflammatory response and improve prognosis, and has good safety, but also can improve hypotension of patients with sepsis shock and deficiency of healthy qi and excess of evil qi, improve hemodynamics, tissue perfusion and blood coagulation function of patients with sepsis shock, and has no obvious influence on liver and kidney functions, and has good safety.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine, specifically relating to a traditional Chinese medicine composition for treating septic cardiomyopathy and its application. Background Technology

[0002] Sepsis is a common and serious complication in patients with acute and critical illnesses such as trauma, shock, and infection. It remains a significant threat to human health, characterized by high morbidity, high mortality, and high treatment costs. Sepsis-induced cardiomyopathy (SIC), also known as sepsis-induced myocardial dysfunction (SIMD) or sepsis-related myocardial injury, is one of the most serious complications and leading causes of death in sepsis, directly impacting prognosis. Studies have shown that the mortality rate for sepsis patients with cardiac dysfunction is as high as 70%, while the mortality rate for those without cardiac dysfunction is only 20%. Effective cardioprotection can improve the prognosis of sepsis patients and reduce their mortality rate. Myocardial injury is the core pathogenesis of sepsis-induced cardiomyopathy, while cardiac dysfunction is the clinical manifestation. Its pathogenesis is complex, and there is currently no specific treatment.

[0003] Modern medicine treats septic cardiomyopathy primarily by addressing the underlying cause of sepsis (anti-infection) combined with hemodynamic support. Traditional Chinese medicine treatment mainly involves injectable Chinese herbal medicines such as Xuebijing injection and Shenfu injection. Currently, there is still a lack of clinically effective traditional Chinese medicine formulas for septic cardiomyopathy. Summary of the Invention

[0004] The first objective of this invention is to provide a traditional Chinese medicine composition for treating septic cardiomyopathy, and the second objective of this invention is to provide the application of this traditional Chinese medicine composition.

[0005] According to a first aspect of the present invention, a traditional Chinese medicine composition for treating septic cardiomyopathy is provided, the main active ingredients comprising, by weight: 10-50 parts ginseng, 5-20 parts rhubarb, 10-30 parts aconite, 5-20 parts coptis, 5-30 parts scutellaria, and 5-20 parts safflower.

[0006] This invention posits that sepsis commonly occurs in the elderly and those with weakened constitutions, or after major surgery. In these cases, the body's vital energy (Qi) is deficient, the defensive Qi (Wei Qi) is weakened, and pathogenic factors (Huang Qi, Yin and Yang) prevail. External pathogens, such as the six evils (wind, cold, heat, dryness, fire, and fire) and other harmful influences, invade the lungs and defensive Qi, further depleting the body's vital energy. These pathogenic factors then enter the blood and affect the pericardium. After the invasion of pathogenic factors, the heart's Yang Qi is damaged, and Yang deficiency weakens its ability to circulate blood. The kidneys are the foundation of Yin and Yang; insufficient kidney Yang Qi prevents the warming of the heart's Yang Qi, leading to further depletion of the heart's Yang Qi. Furthermore, the body cannot properly metabolize fluids into urine for excretion, resulting in edema. Since septic cardiomyopathy occurs when the body's vital energy is already deficient and pathogenic factors prevail, the etiology of septic cardiomyopathy is fundamentally based on the deficiency of the body's vital energy, representing a deficiency in the root and an excess in the branch.

[0007] Sepsis-induced myocardial damage is fundamentally due to a deficiency of the body's vital energy (Qi), representing a deficiency in the root and an excess in the branch, with Yang deficiency being the primary pathogenic factor. Cardiac dysfunction or hemodynamic disturbances in sepsis patients are clinical manifestations of myocardial involvement caused by severe infection. In the early stages of the disease, a strategy of intervention and reversal should be employed based on syndrome differentiation, including methods such as inducing sweating, detoxification, bowel regulation, or blood circulation promotion, treating according to the specific symptoms to prevent further disease progression. Simultaneously, since most patients with septic cardiomyopathy experience deficiency of heart Qi or heart Yang, leading to decreased systolic and / or diastolic function, and even hemodynamic disturbances such as shock, it is necessary to replenish deficiency throughout the treatment, protect the body's vital energy, tonify heart Qi (Yang), improve cardiac function, stabilize hemodynamic parameters, and thus improve the prognosis of patients with septic cardiomyopathy.

[0008] This invention adopts the principle of supporting the body's resistance and eliminating pathogenic factors, and the treatment method of warming the yang, detoxifying, and promoting blood circulation to remove blood stasis. In the traditional Chinese medicine composition of this invention, ginseng is the principal ingredient, which enters the heart meridian, greatly replenishes the original qi, nourishes the internal organs, and directly improves the cardiac function of patients with sepsis.

[0009] Rhubarb and aconite are used as assistant herbs. Rhubarb purges stagnation, clears heat, detoxifies, cools the blood, stops bleeding, invigorates blood circulation, removes blood stasis, and clears damp-heat. When used with ginseng, they complement each other, purging turbidity, eliminating toxins, and tonifying qi and supporting the body's resistance. When rhubarb and ginseng are used together, rhubarb removes hard accumulations without harming the body's vital energy, while ginseng replenishes the body's vital energy without tonifying pathogenic factors. The synergistic effect of rhubarb and ginseng, one tonifying and the other purging, invigorates blood circulation and detoxifies. Aconite restores yang, rescues from collapse, assists yang, tonifies fire, dispels cold, and relieves pain, assisting ginseng in exerting its therapeutic effects.

[0010] When rhubarb is combined with aconite, the aconite, warming and invigorating yang qi and dispelling yin cold, is effective in treating cold-induced stagnation syndromes. However, because aconite is warming, improper use can lead to dryness and damage to body fluids, causing cold stagnation to evolve into dryness stagnation. Therefore, combining aconite with rhubarb is a medicinal pair. Rhubarb guides aconite directly to the affected area while preventing it from drying out, and rhubarb is restrained by aconite to avoid exacerbating cold. This combination works synergistically to treat cold-induced stagnation syndromes. Simultaneously, the combination of rhubarb and aconite can also treat syndromes where the pathogenesis involves both internal accumulation of pathogenic heat and internal retention of cold. Rhubarb is used to purge heat, while aconite warms yang, working synergistically to treat yang deficiency with internal cold and internal accumulation of pathogenic heat.

[0011] Coptis chinensis and Scutellaria baicalensis are used as adjuvant herbs. Coptis chinensis clears heat and dries dampness, purges fire and detoxifies; Scutellaria baicalensis clears heat and dries dampness, purges fire and detoxifies, cools blood and stops bleeding. The adjuvant herbs have opposite properties to the principal and assistant herbs, thus restraining any imbalances in their therapeutic effects. At the same time, ginseng tonifies the spleen and replenishes qi, preventing the bitter and cold properties of Coptis chinensis and Scutellaria baicalensis from harming the stomach. The combination of these herbs promotes the flow of qi through the interaction of pungent and bitter herbs, clears heat from the upper body, warms the middle body, and tonifies the lower body, thus harmonizing the spleen and stomach.

[0012] Safflower is used as the guiding herb to invigorate blood circulation, regulate menstruation, dispel blood stasis, and relieve pain. It enters the heart meridian and acts as a guiding herb to ensure that the medicinal effects reach the whole body, invigorate blood circulation, remove blood stasis, improve microcirculation, and enable the whole formula to exert its therapeutic effects better and faster.

[0013] This formula aligns with the pathogenesis of sepsis characterized by deficiency of vital energy and excess of pathogenic factors, and also corresponds to the symptoms of septic cardiomyopathy involving deficiency of heart yang and blood stasis with toxic accumulation. The entire formula combines warming and cooling properties, balancing attack and tonification, addressing both the root cause and the symptoms. It clears heat and detoxifies, cutting off the further transmission of external pathogens, while also warming heart yang, unblocking the meridians and removing blood stasis, supporting the body's vital energy to expel pathogens. The entire formula works synergistically to detoxify, clear the bowels, warm yang, and invigorate blood, effectively improving myocardial function while treating sepsis.

[0014] The raw materials for this formula come from the following sources:

[0015] Ginseng: The dried root of Panax ginseng CAMey., a plant belonging to the Araliaceae family. It enters the spleen, lung, and heart meridians. It is sweet and slightly bitter, and neutral in nature.

[0016] Rhubarb: The dried root and rhizome of *Rheum palmatum* L., *Rheum tanguticum* Maxim. ex Balf., or *Rheum officinale* Baill., all belonging to the Polygonaceae family. It enters the spleen, stomach, large intestine, liver, and pericardium meridians. It is bitter and cold in nature.

[0017] Aconite: A processed product of the rootlets of *Aconitum carmichaeli* Debx., a plant in the Ranunculaceae family. It enters the Heart, Kidney, and Spleen meridians. It is pungent, sweet, and very hot in nature.

[0018] Coptis chinensis: The dried rhizome of *Coptis chinensis* Franch., *Coptis deltoidea* C.Y.Cheng et Hsiao, or *Coptis teeta* Wall., all belonging to the Ranunculaceae family. It enters the Heart, Spleen, Stomach, Liver, Gallbladder, and Large Intestine meridians. It is bitter and cold in nature.

[0019] Scutellaria baicalensis Georgi, a plant in the Lamiaceae family. It enters the Lung, Gallbladder, Spleen, Large Intestine, and Small Intestine meridians. It is bitter and cold in nature.

[0020] Safflower: The dried flower of *Carthamus tinctorius* L., a plant in the Asteraceae family. It enters the Heart and Liver meridians. It is pungent and warm in nature.

[0021] In some embodiments, the main active ingredients, by weight, include: 20 parts ginseng, 6 parts rhubarb, 15 parts aconite, 6 parts coptis, 6 parts scutellaria, and 10 parts safflower.

[0022] In some embodiments, the main active ingredients, by weight, include: 15 parts ginseng, 5 parts rhubarb, 10 parts aconite, 5 parts coptis, 5 parts scutellaria, and 10 parts safflower.

[0023] In some embodiments, the main active ingredients, by weight, include: 15 parts ginseng, 10 parts rhubarb, 10 parts aconite, 10 parts coptis, 10 parts scutellaria, and 10 parts safflower.

[0024] In some embodiments, the main active ingredients, by weight, include: 20 parts ginseng, 10 parts rhubarb, 15 parts aconite, 10 parts coptis, 15 parts scutellaria, and 5 parts safflower.

[0025] In some embodiments, the main active ingredients, by weight, include: 50 parts ginseng, 20 parts rhubarb, 30 parts aconite, 20 parts coptis, 30 parts scutellaria, and 20 parts safflower.

[0026] The dosage of each component in the traditional Chinese medicine composition of the present invention can be adjusted according to the symptoms. In addition to the dosage ratios listed above, other ratios within the above dosage range can also be used, which will not be listed here.

[0027] In some implementations, the aconite is processed aconite.

[0028] In some embodiments, medically acceptable excipients are also included. Specifically, the excipients used are carriers or excipients commonly used in the art, including but not limited to starch, lactose, glucose, sodium carboxymethyl cellulose, ethyl cellulose and methyl cellulose, malt, gelatin, polyols (such as propylene glycol, glycerol, mannitol), tablets, etc., to help improve the stability or activity of the drug, or to produce an acceptable taste or odor when taken orally.

[0029] In some embodiments, the dosage form of the traditional Chinese medicine composition is a decoction, granules, powder, tablets, pills, or capsules. This facilitates clinical administration.

[0030] According to a second aspect of the present invention, the use of the above-described traditional Chinese medicine composition for treating septic cardiomyopathy in the preparation of a medicament for treating sepsis is provided.

[0031] According to a third aspect of the present invention, the use of the above-described traditional Chinese medicine composition for treating septic cardiomyopathy in the preparation of a medicament for treating septic cardiomyopathy is provided.

[0032] According to a fourth aspect of the present invention, the above-described traditional Chinese medicine composition for treating septic cardiomyopathy is provided for use in the preparation of a medicine for treating septicemia with yang deficiency and toxic stasis syndrome or septic shock with deficiency of vital energy and excess of pathogenic factors. Attached Figure Description

[0033] Figure 1 This is a flowchart of the clinical study of this invention.

[0034] Figure 2 This is a comparison of the main outcome indicators before and after treatment in the first clinical study of this invention.

[0035] Figure 3 This is a comparison of some cardiac function indicators after treatment in the first clinical study of this invention.

[0036] Figure 4 This is a comparison of inflammatory markers after treatment in the first clinical study of this invention.

[0037] Figure 5 This is a flowchart of the second clinical study of the present invention.

[0038] Figure 6 This is a comparison of the mean arterial pressure after treatment and the difference before and after treatment in the second clinical study of this invention.

[0039] Figure 7 This is a comparison of CO and Lac before and after treatment in Clinical Study 2 of this invention.

[0040] Figure 8 This is a comparison of the survival time of rats in the CLP group and rats in different doses of WY in the animal experiments of this invention.

[0041] Figure 9 This is a comparison of myocardial injury markers between rats in the CLP group and rats in different doses of WY in animal experiments of this invention.

[0042] Figure 10 This is a comparison of inflammatory factors between CLP group rats and different doses of WY group rats in the animal experiments of this invention.

[0043] Figure 11 This is a comparison of the survival time of rats in different groups in the animal experiments of this invention.

[0044] Figure 12 These are echocardiograms of rats from various groups in the animal experiments of this invention.

[0045] Figure 13 This is a comparison of echocardiographic parameters of different groups of rats in the animal experiments of this invention.

[0046] Figure 14This is a comparison of myocardial injury markers in different groups of rats in the animal experiments of this invention.

[0047] Figure 15 This is a comparison of inflammatory factors in different groups of rats in the animal experiments of this invention.

[0048] Figure 16 This is a comparison of oxidative stress factors in different groups of rats in the animal experiments of this invention.

[0049] Figure 17 This is a comparison of the expression of autophagy-related proteins in different groups of rats in the animal experiments of this invention.

[0050] Figure 18 These are pathological sections of rat myocardium from various groups in the animal experiments of this invention.

[0051] Figure 19 This is a comparison of autophagosomes in the myocardial endothelial cells of rats in animal experiments of this invention. Detailed Implementation

[0052] The present invention will be further described in detail below with reference to specific embodiments. It is worth noting that the following embodiments are only for better explanation of the content of the present invention and do not limit the scope of protection of the present invention. Process steps not disclosed in the embodiments are prior art. Unless otherwise specified, all raw materials used in the present invention are commercially available.

[0053] Example 1

[0054] The traditional Chinese medicine decoction for treating septic cardiomyopathy in this embodiment is prepared from the following components: ginseng 20g, rhubarb 6g, prepared aconite root 15g, coptis root 6g, scutellaria root 6g, and safflower 10g.

[0055] The preparation method is to prepare the decoction according to conventional methods in this field.

[0056] For example, its preparation method includes the following steps:

[0057] Wash the herbs, add 10 times their weight of water until the water level is even with the herbs, soak for 30 minutes, bring to a boil over high heat, then simmer over low heat for 40 minutes and extract the juice; add 8 times their weight of water to the dregs until the water level is even with the dregs, simmer again and extract the juice, then mix the juices obtained from the two simmerings to obtain about 100ml of Chinese herbal decoction.

[0058] One dose is 100ml. When administering the medication to the patient, one dose is given daily, orally or via nasogastric tube.

[0059] Example 2

[0060] The granules for treating septic cardiomyopathy in this embodiment are prepared from the following components: ginseng 15g, rhubarb 5g, prepared aconite root 10g, coptis root 5g, scutellaria root 5g, and safflower 10g.

[0061] The preparation method is to prepare granules according to conventional methods in this field.

[0062] Example 3

[0063] The powder for treating septic cardiomyopathy in this embodiment is prepared from the following components: ginseng 15g, rhubarb 10g, prepared aconite root 10g, coptis root 10g, scutellaria root 10g, and safflower 10g.

[0064] The preparation method is to prepare the powder according to conventional methods in this field.

[0065] Example 4

[0066] The tablet for treating septic cardiomyopathy in this embodiment is prepared from the following components: ginseng 20g, rhubarb 10g, prepared aconite root 15g, coptis root 10g, scutellaria root 15g, and safflower 5g.

[0067] The tablets are prepared according to conventional methods in the field.

[0068] Example 5

[0069] The pill for treating septic cardiomyopathy in this embodiment is prepared from the following components: ginseng 50g, rhubarb 20g, prepared aconite root 30g, coptis root 20g, scutellaria root 30g, and safflower 20g.

[0070] The preparation method is to prepare pills according to conventional methods in this field.

[0071] The following clinical and animal experiments were conducted to investigate the therapeutic effects of the traditional Chinese medicine composition for treating septic cardiomyopathy (hereinafter referred to as Wen Yang Jie Du Fang) of the present invention on sepsis and septic cardiomyopathy.

[0072] I. Clinical Research

[0073] This study evaluates the efficacy of the Wenyang Jiedu formula in patients with sepsis.

[0074] This study was a prospective, single-blind, randomized controlled trial. Patients with sepsis and TCM diagnosis of Yang deficiency and toxic stasis were randomly assigned to a control group and a treatment group. The control group received standard Western medicine treatment plus a placebo (note: the placebo was an equivalent dose of warm water), while the treatment group received standard Western medicine treatment plus a TCM decoction for treating septic cardiomyopathy prepared in Example 1 (referred to as Wen Yang Jie Du Fang decoction). Dosage and administration: one dose daily, approximately 100ml, orally / nasally. Treatment lasted for 5 days, with follow-up until 28 days. The Sequential Organ Failure Assessment (SOFA) score, Acute Pathophysiology and Long-Term Health Evaluation II (APACHE II) score, 28-day mortality, and tissue perfusion were compared between the two groups. The flowchart of this study is shown below. Figure 1 As shown.

[0075] The TCM diagnostic criteria used in this study were based on the "Guidelines for the Treatment of Severe Sepsis / Septic Shock in China" promulgated in 2014. The TCM syndrome differentiation of sepsis can be divided into four categories: toxic heat syndrome, obstruction of bowel function syndrome, blood stasis syndrome, and acute deficiency syndrome. According to previous epidemiological surveys and literature reviews, clinical sepsis syndromes are mainly characterized by concurrent symptoms. This study selected the Yang deficiency and toxic stasis syndrome, specifically manifested as: fatigue and weakness, shortness of breath and reluctance to speak, pain described as needle-like or knife-like, fixed in location, or often worsening at night; some patients may present with lumps and bleeding; pale or purplish tongue, or with ecchymosis and petechiae; varicose veins under the tongue; deep and thready pulse, or deep and slow pulse, or thready and hesitant pulse; or cold sweats, cold extremities, flushed face, cyanotic lips; or high fever, irritability, and delirium.

[0076] Inclusion criteria were: (i) meeting the diagnostic criteria for sepsis; (ii) being diagnosed with Yang deficiency and blood stasis syndrome according to traditional Chinese medicine; (iii) being aged 18-85 years; (iv) having normal gastrointestinal function and being able to be fed via nasogastric tube or orally; and (v) being willing to participate in this study and signing an informed consent form.

[0077] Exclusion criteria were: (i) pregnant or lactating women; (ii) those with severe liver or kidney dysfunction or acute cerebrovascular disease before enrollment; (iii) those with allergic constitution or allergy to the main drug components of the warming and detoxifying formula of this invention; and (iv) those who participated in other clinical studies at the same time or within the past 6 months.

[0078] The dropout criteria were: (i) the subject was unwilling to continue the clinical trial and requested to withdraw from the clinical trial or to abandon treatment; and (ii) follow-up information was missing. Data from dropouts were excluded and not included in statistical analysis.

[0079] The exclusion criteria were: (i) mis-admission or misdiagnosis; (ii) only baseline data were available, making it impossible to identify the efficacy, and there was no valid data; (iii) poor subject compliance, failure to adhere to or serious violation of the trial design protocol; (iv) failure to use medication as prescribed, making it impossible to judge the trial effect, or incomplete data that hindered the judgment of efficacy or safety.

[0080] The criteria for terminating the trial are: (i) a serious adverse event occurs during the trial; (ii) serious complications or comorbidities arise from the trial and the subject is not suitable to continue the trial; (iii) the subject's condition deteriorates rapidly and requires immediate treatment; (iv) the circumstances listed in the exclusion criteria occur; or (v) other circumstances arise that the researchers determine require termination of the study.

[0081] A total of 71 patients were included, with 34 in the control group and 37 in the treatment group. General information of the two groups is shown in Table 1.

[0082] Table 1 Comparison of general information before treatment

[0083]

[0084]

[0085] As shown in Table 1, there were no statistically significant differences in the baseline data of the two groups of patients before treatment (P>0.05), indicating that they were comparable.

[0086] Table 2 shows a comparison of the circulatory indicators of the two groups of patients before treatment.

[0087] Table 2 Comparison of pre-treatment circulatory indicators

[0088] index Control group (n=34) Treatment group (n=37) <![CDATA[Z / t / x 2 ]]> P Lac (mmol / L) 2.22±0.95 1.94±0.74 1.362 0.178 <![CDATA[PaCO2(mmHg)]]> 42.65±5.92 41.97±5.30 0.506 0.614 MAP (mmHg) 79.44±11.47 80.76±9.77 -0.521 0.604 Blood perfusion (PU) 96.12±12.26 96.83±9.72 -0.270 0.788

[0089] Note: P > 0.05 indicates no statistically significant difference between the two groups. Both the control and treatment groups showed normality in Lac (SW test, P values ​​were 0.09 and 0.078, respectively), as determined by a two-sample t-test. Both the control and treatment groups showed normality in PaCO2 (SW test, P values ​​were 0.375 and 0.288, respectively), as determined by a two-sample t-test. Both the control and treatment groups showed normality in MAP (SW test, P values ​​were 0.951 and 0.271, respectively), as determined by a two-sample t-test. Both the control and treatment groups showed normality in blood perfusion (SW test, P values ​​were 0.926 and 0.603, respectively), as determined by a two-sample t-test.

[0090] As shown in Table 2, there were no statistically significant differences in Lac, PaCO2, MAP, and blood perfusion between the two groups before treatment (P>0.05), indicating that they were comparable.

[0091] Table 3 shows a comparison of cardiac function indicators between the two groups of patients before treatment.

[0092] Table 3 Comparison of cardiac function indicators before treatment

[0093]

[0094] Note: P > 0.05 indicates no statistically significant difference between the two groups. LVEF in both the control and treatment groups did not meet the normality requirement (SW test, P values ​​were 0.012 and 0.002, respectively), so an independent samples rank-sum test was used. FS in both the control and treatment groups met the normality requirement (SW test, P values ​​were 0.366 and 0.063, respectively), so an independent samples t-test was used. LVEDD in both the control and treatment groups met the normality requirement (SW test, P values ​​were 0.597 and 0.434, respectively), so an independent samples t-test was used. LVESD in the control group met the normality requirement, while LVESD in the treatment group did not (SW test, P values ​​were 0.094 and 0.021, respectively). Since neither group fully met the normality requirement, an independent samples rank-sum test was used. CK in both the control and treatment groups did not meet the normality requirement (SW test, P values ​​were both 0.000), so an independent samples rank-sum test was used.

[0095] As shown in Table 3, there were no statistically significant differences in cardiac function indicators between the two groups before treatment (P>0.05), indicating that they were comparable.

[0096] Table 4 shows a comparison of the primary outcome indicators between the two groups of patients after treatment. Figure 2 As shown.

[0097] Table 4 Comparison of major outcome indicators after treatment

[0098] index Control group (n=34) Treatment group (n=37) <![CDATA[Z / t / x 2 ]]> P Lac (mmol / L) 2.57±0.89 1.61±0.56 5.441 <0.001 <![CDATA[PaCO2(mmHg)]]> 40.85±2.50 40.00(39.00,41.00) -2.211 0.027 MAP (mmHg) 74.97±7.99 83.32±9.32 -4.039 <0.001

[0099] Note: P > 0.05 indicates no statistically significant difference between the two groups. Both the control and treatment groups showed normality in Lac (SW test, P values ​​were 0.91 and 0.05, respectively), so an independent samples t-test was used. The control group's PaCO2 showed normality, while the treatment group's PaCO2 did not (SW test, P values ​​were 0.301 and 0.029, respectively). Since neither group fully met normality, an independent samples rank-sum test was used. Both the control and treatment groups showed normality in MAP (SW test, P values ​​were 0.493 and 0.557, respectively), so an independent samples t-test was used.

[0100] From Table 4 and Figure 2 As can be seen, the comparison of the primary outcome indicators between the two groups of patients after treatment showed a statistically significant difference (P < 0.05). This indicates that the traditional Chinese medicine composition of the present invention can significantly improve the primary outcome indicators Lac, PaCO2, and MAP in patients with sepsis.

[0101] Table 5 shows a comparison of cardiac function indicators between the two groups of patients after treatment. The results of comparisons of some cardiac function indicators after treatment are as follows: Figure 3 As shown.

[0102] Table 5 Comparison of cardiac function indicators after treatment

[0103]

[0104]

[0105] Note: P > 0.05 indicates no statistically significant difference between the two groups. LVEF in both the control and treatment groups did not meet the normality requirement (SW test, P values ​​were 0.000 and 0.024, respectively), and a two-sample rank-sum test was used. FS in both the control and treatment groups met the normality requirement (SW test, P values ​​were 0.426 and 0.146, respectively), and a two-sample t-test was used. LVEDD in both the control and treatment groups met the normality requirement (SW test, P values ​​were 0.169 and 0.429, respectively), and a two-sample t-test was used. LVESD in the control group met the normality requirement, while LVESD in the treatment group did not (SW test, P values ​​were 0.562 and 0.03, respectively). Since neither group fully satisfies normality, the independent samples rank-sum test is used. Neither the control nor treatment group CK satisfies normality (SW test, P values ​​both 0.000), so the independent samples rank-sum test is used. Neither the control nor treatment group CK-MB satisfies normality (SW test, P values ​​both 0.000), so the independent samples rank-sum test is used. Neither the control nor treatment group TnT satisfies normality (SW test, P values ​​both 0.000), so the independent samples rank-sum test is used. Neither the control nor treatment group pro-BNP satisfies normality (SW test, P values ​​both 0.000), so the independent samples rank-sum test is used.

[0106] As shown in Table 5, the cardiac function indicators LVEF, LVESD, CK, TnT, and pro-BNP were significantly different between the two groups after treatment (P < 0.05). This indicates that the traditional Chinese medicine composition of the present invention can significantly improve the cardiac function indicators LVEF, LVESD, CK, TnT, and pro-BNP in patients with sepsis.

[0107] The comparison of inflammatory markers between the two groups of patients after treatment is shown in Table 6. Figure 4 As shown.

[0108] Table 6 Comparison of inflammatory markers after treatment

[0109]

[0110] Note: P > 0.05 indicates no statistically significant difference between the two groups. WBC in both the control and treatment groups did not meet the normality requirement (SW test, P values ​​were 0.002 and 0.003, respectively), so a two-sample rank-sum test was used. NEUT in the control group did not meet the normality requirement, while NEUT in the treatment group did (SW test, P values ​​were 0.009 and 0.42, respectively). Since neither group fully met the normality requirement, a two-sample rank-sum test was used. hs-CRP in both the control and treatment groups did not meet the normality requirement (SW test, P values ​​were 0.031 and 0.006, respectively), so a two-sample rank-sum test was used. PCT in both the control and treatment groups did not meet the normality requirement (SW test, P values ​​were both 0.000), so a two-sample rank-sum test was used.

[0111] As shown in Table 6, the inflammatory markers WBC and hs-CRP levels were significantly different between the two groups after treatment (P < 0.05). This indicates that the traditional Chinese medicine composition of the present invention can significantly improve the inflammatory markers WBC and hs-CRP in patients with sepsis.

[0112] Table 7 shows a comparison of the post-treatment scores and health economic indicators between the two groups of patients.

[0113] Table 7 Comparison of post-treatment scores and health economic indicators

[0114]

[0115] Note: P > 0.05 indicates no statistically significant difference between the two groups. The length of hospital stay in both the control and treatment groups did not meet the normality requirement (SW test, P values ​​were 0.006 and 0.000, respectively), and a two-sample rank-sum test was used. The number of organ failures in both the control and treatment groups did not meet the normality requirement (SW test, P values ​​were 0.011 and 0.001, respectively), and a two-sample rank-sum test was used. The mechanical ventilation time in both the control and treatment groups did not meet the normality requirement (SW test, P values ​​were 0.029 and 0.014, respectively), and a two-sample rank-sum test was used. The duration of vasoactive drug use in the control group did not meet the normality requirement, while the duration of vasoactive drug use in the treatment group did meet the normality requirement (SW test, P values ​​were 0.029 and 0.014, respectively). The W test showed P values ​​of 0.045 and 0.110, respectively. Since neither group fully met the normality requirement, the independent samples rank-sum test was used. The SOFA scores of the control group met the normality requirement, while those of the treatment group did not (SW test, P values ​​of 0.139 and 0.000, respectively). Since neither group fully met the normality requirement, the independent samples rank-sum test was used. The APACHEII scores of the control group met the normality requirement, while those of the treatment group did not (SW test, P values ​​of 0.590 and 0.000, respectively). Since neither group fully met the normality requirement, the independent samples rank-sum test was used.

[0116] As shown in Table 7, there were statistically significant differences between the two groups in post-treatment scores and health economic indicators, including 28-day mortality, length of hospital stay, number of organ failures, duration of mechanical ventilation, duration of vasoactive drug use, SOFA score, and APACHE II score (P < 0.05). This indicates that the traditional Chinese medicine composition of the present invention can significantly improve the scores and health economic indicators of sepsis patients, including 28-day mortality, length of hospital stay, number of organ failures, duration of mechanical ventilation, duration of vasoactive drug use, SOFA score, and APACHE II score.

[0117] Table 8 shows the comparison of liver, kidney, and coagulation indicators between the two groups of patients after treatment.

[0118] Table 8 Comparison of liver, kidney, and coagulation indicators after treatment

[0119]

[0120] Note: P > 0.05 indicates that there is no statistically significant difference between the two groups. The control group's SCr met the normality requirement, while the treatment group's SCr did not (SW test, P values ​​were 0.748 and 0.003, respectively). Since neither group fully met the normality requirement, a two-sample rank-sum test was used. The control group's TP did not meet the normality requirement, while the treatment group's TP met the normality requirement (SW test, P values ​​were 0.041 and 0.164, respectively). Since neither group fully met the normality requirement, a two-sample rank-sum test was used. Both the control and treatment groups' TBIL did not meet the normality requirement (SW test, P values ​​were both 0.000), so a two-sample rank-sum test was used. The control group's ALT met the normality requirement, while the treatment group's ALT did not (SW test, P values ​​were 0.075 and 0.000, respectively). Since neither group fully met the normality requirement, a two-sample rank-sum test was used. Both the control and treatment groups' AST did not meet the normality requirement (SW test, P values ​​were 0.000 and 0.001, respectively), so a two-sample rank-sum test was used. The control group's PT met the normality requirement, while the treatment group's PT... The control and treatment groups did not meet the normality requirement (SW test, P values ​​were 0.237 and 0.007, respectively). Since neither group fully met the normality requirement, the independent samples rank-sum test was used. The APTT of both groups met the normality requirement (SW test, P values ​​were 0.089 and 0.098, respectively), and the independent samples t-test was used. The INR of both groups met the normality requirement (SW test, P values ​​were 0.63 and 0.678, respectively), and the independent samples t-test was used. D-dimer levels did not satisfy normality (SW test, P values ​​were all 0.000), so a two-sample rank-sum test was used; Hb levels in both the control and treatment groups satisfied normality (SW test, P values ​​were 0.793 and 0.58, respectively), so a two-sample t-test was used; PLT levels in the control group satisfied normality, while PLT levels in the treatment group did not (SW test, P values ​​were 0.125 and 0.007, respectively), indicating that neither group fully satisfied normality, so a two-sample rank-sum test was used; K... + Satisfying normality, treatment group K + The two groups do not meet the normality requirement (SW test, P values ​​are 0.154 and 0.000 respectively). Since neither group fully meets the normality requirement, the rank-sum test of two independent samples is used.

[0121] As shown in Table 8, there were no statistically significant differences in liver, kidney, and coagulation indicators between the two groups after treatment (P > 0.05). This indicates that the herbal composition of the present invention has no significant effect on liver, kidney, and coagulation indicators.

[0122] In summary, compared with the control group, the treatment group had higher local tissue perfusion, lower 28-day mortality, shorter duration of vasoactive drug use, and fewer organ failures after treatment. Post-treatment levels were also lower in blood lactate (Lac), arterial carbon dioxide partial pressure (PaCO2), creatine kinase (CK), troponin (TnT), pro-BNP, white blood cells (WBC), high-sensitivity C-reactive protein (hs-CRP), APACHE II score, and SOFA score, and higher in mean arterial pressure (MAP) and left ventricular ejection fraction (LVEF) (all P < 0.05). There were no statistically significant differences in coagulation function and liver and kidney function between the two groups after treatment (all P > 0.05). This suggests that the traditional Chinese medicine composition of this invention can significantly improve circulation and cardiac function in patients with sepsis due to Yang deficiency and blood stasis syndrome, reduce inflammatory response, improve prognosis, and has good safety.

[0123] II. Clinical Research II

[0124] This study investigates the effects of the Wen Yang Jie Du formula on hemodynamics and vascular endothelial function in patients with septic shock of the deficiency-excess type.

[0125] This prospective, randomized controlled trial included patients with septic shock (deficiency of vital energy and excess of pathogenic factors) admitted to the Intensive Care Unit of a hospital in Guangdong Province between February 2022 and February 2023. Patients were randomly assigned to the experimental group and the control group using a random number table generated by SPSS 26 software. The control group received standard treatment according to the 2021 International Guidelines for the Management of Sepsis and Septic Shock. The experimental group received the same treatment as the control group, plus the traditional Chinese medicine decoction for treating septic cardiomyopathy prepared in Example 1 (referred to as Wen Yang Jie Du Fang decoction). Dosage and administration: one dose daily, approximately 100 ml, orally / nasally. Both groups were treated for 5 days, with a 5-day observation period. Hemodynamics and vascular endothelial function, tissue perfusion parameters, coagulation function, inflammatory factors, liver and kidney function, myocardial injury markers, 28-day mortality, APACHE II score, and SOFA score were observed before and after treatment. Statistical analysis was performed on the results to evaluate the efficacy of the Wen Yang Jie Du formula in patients with septic shock of the deficiency-excess type. The flowchart of this study is shown below. Figure 5 As shown.

[0126] The TCM diagnostic criteria used in this study were as follows: The patient met the criteria for septic shock with a deficiency of vital energy and excess of pathogenic factors, referring to the 2021 "TCM Clinical Diagnosis and Treatment Terminology Part 2: Syndromes" and the 2019 "Expert Consensus on the Integrated Traditional Chinese and Western Medicine Diagnosis and Treatment of Septic Shock": ① Persistent high or low fever, lethargy, pale complexion, and emaciation; or lethargy, sallow or pale complexion, and emaciated body; red tongue with yellow-white or yellow or grayish-greasy tongue coating, and a thready, rapid, weak pulse, or a thready, weak pulse, etc. ② Persistent high or low fever, sudden pale complexion, weak breathing, profuse cold sweat, cold extremities, scanty urine, snoring, delirium, limb weakness, or even coma; or delirium, weak breathing, scanty urine, accompanied by wind-heat or blood-heat symptoms; pale tongue with moist tongue coating, and a faint, weak, or floating pulse, etc.

[0127] A comprehensive evaluation by the principal investigator and TCM physicians with the title of attending physician or above is required. If two or more of ① or ② are met, the patient can be diagnosed with septic shock due to deficiency of vital energy and excess of pathogenic factors.

[0128] The inclusion criteria were: (1) Age 18-85 years; no gender requirement. (2) In Western medicine, the patient met the definition and diagnostic criteria for septic shock in the 2016 Third International Consensus: New Definition of Sepsis and Septic Shock, namely: ① Diagnosis of sepsis: Infected or suspected infected patients with a Sequential Organ Failure Assessment (SOFA) score ≥2 points from baseline. ② Diagnosis of septic shock: Based on the diagnosis of sepsis, the patient still has persistent hypotension after adequate volume resuscitation and requires vasoactive drugs to maintain a mean arterial pressure (MAP) ≥65 mmHg and a serum lactate concentration >2 mmol / L. (3) According to the TCM syndrome differentiation criteria, the patient met the syndrome of septic shock with deficiency of vital energy and excess of pathogenic factors. (4) The patient voluntarily participated in this study and signed an informed consent form. (5) The patient was willing to accept TCM treatment and cooperate with relevant examinations.

[0129] Exclusion criteria are: (1) Patients with acute myocardial infarction. (2) Patients whose SOFA score is ≥12 points for three items: coagulation function (platelet count), kidney function (creatinine), and liver function (bilirubin), specifically: ① Patients with severe bleeding tendency or active bleeding (platelet count <21×10). 9 (2) Individuals with severe renal insufficiency (creatinine > 440 μmol / L); (3) Individuals with severe hepatic impairment (bilirubin > 204 μmol / L). (4) Individuals who died or were discharged from the ICU (< 5 days) before the eligibility assessment. (5) Individuals who are allergic to the relevant investigational drug. (6) Individuals who participated in other drug trials within 3 months prior to the trial. (7) Pregnant or lactating women, individuals with mental disorders, or individuals without legal capacity.

[0130] A total of 40 patients were included, with 20 in the experimental group and 20 in the control group. The baseline general information of the two groups before treatment is shown in Table 9.

[0131] Table 9. Baseline comparison of general information before treatment [M(P25,P75) / x±S]

[0132]

[0133]

[0134] Note: Gender and disease are expressed as a percentage of cases. # indicates the use of nonparametric rank-sum test, and the rest use chi-square test.

[0135] As shown in Table 9, there were no statistically significant differences in baseline age, gender, underlying medical history, and infection site between the two groups of patients before treatment (P > 0.05), indicating that they were comparable.

[0136] Table 10 shows the baseline comparison of hemodynamic parameters between the two groups of patients before treatment.

[0137] Table 10. Baseline comparison of hemodynamic parameters before treatment [M(P25, P75) / x±S]

[0138]

[0139]

[0140] Note: *Indicates the use of independent samples t-test; otherwise, use nonparametric rank-sum test.

[0141] As shown in Table 10, there were no statistically significant differences in hemodynamic parameters between the two groups before treatment (P > 0.05), indicating comparability.

[0142] Table 11 shows the baseline comparison of liver and kidney function and myocardial injury markers between the two groups of patients before treatment.

[0143] Table 11. Baseline comparison of liver and kidney function and myocardial injury markers before treatment [M(P25, P75)]

[0144]

[0145] As shown in Table 11, there were no statistically significant differences in liver and kidney function and myocardial injury markers between the two groups before treatment (P>0.05), indicating that they were comparable.

[0146] Table 12 shows a comparison of mean arterial pressure (MAP) and the difference before and after treatment between the two groups of patients. Figure 6 As shown.

[0147] Table 12 Comparison of MAP after treatment [M(P25,P75) / x±S]

[0148]

[0149] Note: *Indicates the use of independent samples t-test; otherwise, use nonparametric rank-sum test.

[0150] As shown in Table 12, the mean arterial pressure (MAP) of the two groups after treatment and the difference in MAP before and after treatment were statistically significant (P < 0.05). This indicates that the traditional Chinese medicine composition of the present invention can significantly improve the mean arterial pressure in patients with sepsis.

[0151] Table 13 shows a comparison of hemodynamic and tissue perfusion parameters between the two groups after treatment, and Table 14 shows a comparison of the differences in hemodynamic and tissue perfusion parameters before and after treatment between the two groups. The comparisons of CO and Lac before and after treatment between the two groups are shown below. Figure 7 As shown.

[0152] Table 13 Comparison of hemodynamic and tissue perfusion parameters after treatment [M(P25,P75) / x±S]

[0153]

[0154]

[0155] Note: *Indicates the use of independent samples t-test; otherwise, use nonparametric rank-sum test.

[0156] As shown in Table 13, the hemodynamic and tissue perfusion parameters CO, CVP, and Lac were significantly different between the two groups after treatment (P < 0.05). This indicates that the traditional Chinese medicine composition of the present invention can significantly improve the hemodynamic and tissue perfusion parameters CO, CVP, and Lac in patients with sepsis.

[0157] Table 14 Comparison of hemodynamic and tissue perfusion parameters before and after treatment [M(P25,P75) / x±S]

[0158] Experimental group (n=20) Control group (n=20) Z / t p-value CO (L / min) 1.20(0.91,1.60) 0.90(-0.04,1.28) -2.480 0.013 <![CDATA[CI[L / (min·m 2 )]]]> 0.60(0.33,0.95) 0.55(0.10,0.70) -0.884 0.377 <![CDATA[CVP * (cmH2O)]]> 3.75±2.85 2.80±1.77 1.269 <![CDATA[0.214 * ]]> <![CDATA[SVRI * (Pa·s / m 3 )]]> 204.00±270.25 229.50±350.59 -0.258 <![CDATA[0.798 * ]]> Lac (mmol / L) 1.55(1.00,2.38) 1.45(0.90,2.70) -0.027 0.978

[0159] Note: *Indicates the use of independent samples t-test; otherwise, use nonparametric rank-sum test.

[0160] As shown in Table 14, the difference in hemodynamics and tissue perfusion index CO between the two groups before and after treatment was statistically significant (P < 0.05). This indicates that the traditional Chinese medicine composition of the present invention can significantly improve cardiac output in patients with sepsis.

[0161] The differences in liver and kidney function and myocardial injury markers before and after treatment in the two groups of patients are shown in Table 15.

[0162] Table 15 Comparison of differences in liver and kidney function and myocardial injury markers before and after treatment [M(P25,P75)]

[0163]

[0164] As shown in Table 15, the differences in liver and kidney function and myocardial injury markers before and after treatment between the two groups were P>0.05, indicating no statistically significant difference. This suggests that the herbal composition of the present invention has no significant effect on the liver and kidney function or myocardial tissue of the patients.

[0165] In summary, compared with the control group, the primary outcome measure, mean arterial pressure (MAP), was significantly increased in the experimental group after treatment (P<0.05); secondary outcome measures, cardiac output (CO), central venous pressure (CVP), and blood lactate (Lac), were significantly improved (P<0.05); no significant differences were observed in other hemodynamic parameters, vascular endothelial function indicators, inflammatory factors, coagulation function, myocardial injury markers, and safety indicators such as liver and kidney function. No adverse events were recorded. This suggests that the traditional Chinese medicine composition of the present invention can significantly improve hypotension, hemodynamics, tissue perfusion, and coagulation function in patients with septic shock exhibiting both deficiency and excess syndromes; and it has no significant effect on liver and kidney function, demonstrating good safety.

[0166] III. Animal Experimental Research

[0167] This study investigates the mechanism by which the Wen Yang Jie Du formula regulates autophagy in myocardial endothelial cells and improves cardiac function in rats that have undergone cecal ligation and perforation.

[0168] 1. CLP modeling

[0169] A classic SD rat sepsis model was established using cecal ligation (CLP). The CLP rat model was established as follows: After anesthesia with chloral hydrate (0.3 ml / 100 g), a 2-3 cm incision was made in the anterior midline of the abdomen. The mesentery and cecum were freed, and the root of the cecum was ligated with a 30-gauge silk suture in a circular motion. After ligation, the intestinal pathway was carefully checked to ensure it remained intact. Two punctures were made on the serosa of the cecal wall opposite the mesentery using an 18-gauge needle, approximately 1 cm apart. The intestinal tract was gently squeezed to allow feces to overflow from the puncture points. The intestinal tract was then repositioned, and the abdomen was closed layer by layer by suture. Postoperatively, physiological saline (5 ml / 100 g) was injected subcutaneously into the abdominal wall, and fluid resuscitation was performed every 12 hours. Postoperatively, rats were allowed free access to food and water.

[0170] 2. Grouping

[0171] Seventy-five specific pathogen-free (SPF) grade male SD rats were selected. The rats were fed a standard diet with free access to food and water. Those exhibiting normal activity after one week of acclimatization under standard feeding conditions were included in the experiment. The rats were randomly assigned to groups using the PEMS 3.1 statistical software package. After weighing and numbering by weight, the rats were randomly divided into four groups: sham-operated group (Sham group), sepsis model group (CLP group), rapamycin group (RAP group), N-acetylcysteine ​​group (NAC group), and Wenyang Jiedu Detoxification Formula group (WY group), with 15 rats in each group.

[0172] 3. Administration

[0173] Sham group: A sham surgery was performed by intraperitoneal injection of normal saline. Two hours after the operation, 0.9% normal saline was administered by gavage, and thereafter every 12 hours.

[0174] CLP group: 0.9% saline (5 mL / 100 g) was injected subcutaneously into the abdominal wall 30 min after CLP modeling, and the drug was administered once every 12 h.

[0175] RAP group: Rapamycin was injected into the tail vein 10 minutes after CLP modeling at a concentration of 2 μg / kg body weight, once every 12 hours.

[0176] NAC group: 10 minutes after CLP modeling, N-acetylcysteine ​​(NAC), an oxygen free radical scavenger, was injected into the tail vein at a concentration of 150 mg / kg body weight, once every 12 hours.

[0177] WY Group: Rats in the WY group were randomly divided into two groups: the low-dose group of Wen Yang Jie Du Fang (WY-L group) and the medium-dose group of Wen Yang Jie Du Fang (WY-M group), with 7 rats in the WY-L group and 8 rats in the WY-M group. Two hours after CLP modeling, rats were administered the traditional Chinese medicine decoction for treating septic cardiomyopathy prepared in Example 1 by gavage. The concentration of the decoction in the WY-L group was 0.675 g / ml per kilogram of body weight, and the concentration in the WY-M group was 1.35 g / ml per kilogram of body weight, administered every 12 hours.

[0178] The rats in each group were administered the drug for 3 consecutive days.

[0179] It should be noted that the timing and method of drug administration after rat modeling are determined based on the intervention drug for each group. Since CLP modeling involves cecal ligation, which affects the rats' gastrointestinal function, the WY group rats were administered the drug via gavage 2 hours after CLP modeling to minimize the impact of gastrointestinal function on drug absorption. Intravenous administration, however, does not require consideration of gastrointestinal function; therefore, the RAP and NAC groups could begin administration 10 minutes after modeling. Subcutaneous injection via the abdominal wall is the standard fluid resuscitation method for CLP rats; therefore, the CLP group rats underwent subcutaneous injection via the abdominal wall.

[0180] 4. Observation Indicators

[0181] The main observation indicators include:

[0182] (1) Physical indicators

[0183] The physiological state, stimulus response, activity level, and survival rate of rats were observed from the onset of injury to various time points (1h, 2h, 3h, 5h, 7h, 12h, 18h, 24h, 48h, 72h), including respiration, heart rate, body temperature, fecal frequency, and fecal quality.

[0184] (2) Pathological observation of endothelial cell and oxidative stress cell infiltration.

[0185] (3) Cardiac ultrasound

[0186] Using an ultra-high resolution small animal ultrasound imaging system (Vevo2100, Visual Sonies, Canada), rats were anesthetized by inhalation of a mixture of isoflurane and oxygen. The anesthetized rats were then placed on a 37°C constant-temperature mat and continuously inhaled the anesthetic. The extremities of the rats were connected to four electrodes using conductive gel and secured with adhesive tape. An ultrasound coupling agent was applied to the rat's anterior chest area, and echocardiography was performed using an RMV707 B-mode high-frequency ultrasound probe placed on the left chest. After obtaining an ideal parasternal left ventricular long-axis two-dimensional image, the left ventricular outflow tract width and heart rate were measured at the papillary muscle level. At the short-axis level, after obtaining a two-dimensional image, an M-mode echocardiogram was obtained with the M-mode sampling line perpendicular to the interventricular septum and the left ventricular posterior wall at the papillary muscle level. The left ventricular diameter (LVID) and left ventricular posterior wall thickness (LVP) at end-diastole and end-systole were measured. The sectional plane was moved from the left ventricular long axis level to the aortic long axis level. The probe was adjusted and the patient tilted downwards to ensure the probe angle was as consistent as possible with the aortic blood flow angle, controlled within 60°. The sampling port was placed at the valve tip to obtain the aortic blood flow spectrum (VTI). Dynamic images of 10 cardiac cycles were continuously acquired from four different sections. The scanning speed for B-mode ultrasound, M-mode ultrasound, and blood flow Doppler was 800 mm / s. The images were stored and then analyzed offline.

[0187] Various systolic and diastolic parameters of cardiac function, such as left ventricular end-diastolic volume (LVEDV), left ventricular ejection fraction (LVEF%), left ventricular fractional shortening (LVFS%), stroke volume (SV), cardiac output (CO), and E / A ratio, are calculated by the built-in software. The raw data for each group are the average of three consecutive cardiac cycles. The operation and analysis of the ultrasound examination are all performed by professionals.

[0188] (4) The ROS level of cells was detected by DCFH-DA staining.

[0189] (5) Use enzyme-linked immunosorbent assay (ELISA) to detect the levels of relevant molecules in the blood, including the levels of oxidative stress-related molecules such as MDA, SOD, and GSH, as well as the levels of endothelial function markers such as nitric oxide (NO), sFLT1, and ET-1, and the levels of inflammatory mediators such as IL-10, IL-6, and TNF-α, according to the kit steps.

[0190] (6) Detection of protein expression in endothelial cells using Western blotting.

[0191] Western blotting was used to detect the expression of oxidative stress-related proteins such as MDA, SOD, and GSH, as well as autophagy-related proteins LC3 and Beclin1 in cells.

[0192] 5. Experimental Results

[0193] The survival time comparison results of rats in the CLP group and rats in different doses of WY groups are as follows: Figure 8 As shown. From Figure 8 It can be seen that the mortality rate of rats in the CLP group was significantly higher than that in the Sham group, while the mortality rates of rats in the WY-L and WY-M groups were significantly lower than those in the CLP group, with the mortality rate of rats in the WY-L group being even lower than that in the WY-M group.

[0194] The results of the comparison of myocardial injury markers (CK-MB, LDH, α-HBD, cTnT) between CLP group rats and rats in different doses of WY group are as follows: Figure 9 As shown. From Figure 9 It can be seen that the myocardial injury markers CK-MB, α-HBD, and cTnT in the CLP group rats were significantly higher than those in the Sham group, while the levels of CK-MB, α-HBD, and cTnT in the WY-L and WY-M groups were significantly lower than those in the CLP group, with the WY-M group showing a greater decrease.

[0195] The results of the comparison of inflammatory factors (IL-6, IL-10, IL-18) between the CLP group rats and rats in different doses of WY group are as follows: Figure 10 As shown. From Figure 10 It can be seen that the levels of inflammatory factors IL-6, IL-10, and IL-18 in the CLP group rats were significantly higher than those in the Sham group, while the levels of inflammatory factors IL-6 and IL-18 in the WY-L and WY-M groups were significantly lower than those in the CLP group.

[0196] The results of the comparison of survival time of rats in each group are as follows: Figure 11 As shown. From Figure 11 It can be seen that the mortality rate of rats in the CLP group was significantly higher than that in the Sham group, while the mortality rates of rats in the WY and NAC groups were significantly lower than those in the CLP group, with the WY group showing a lower mortality rate than the NAC group. This suggests that both the Wen Yang Jie Du formula and the positive control drug NAC can effectively reduce the mortality rate of rats in the CLP sepsis model, with the Wen Yang Jie Du formula showing a more significant effect.

[0197] Echocardiography of rats in each group as follows Figure 12 As shown. From Figure 12It can be seen that, compared with the Sham group, the cardiac systolic function of rats in the CLP group was worse and the EF and CO decreased at 18 hours and 72 hours after modeling; compared with the CLP group, the cardiac systolic function of rats in the WY group and NAC group was improved.

[0198] The comparison results of echocardiographic parameters (CO, SV, EF, FS) of rats in each group are as follows: Figure 13 As shown. From Figure 13 It can be seen that when the intervention was carried out at 18h, the CO, SV, EF, and FS of rats in the CLP group were significantly lower than those in the Sham group, while the CO and SV of rats in the WY group were significantly higher than those in the CLP group, indicating that cardiac function was improved.

[0199] The comparison results of myocardial injury markers (BNP, cTnT) in each group of rats are as follows: Figure 14 As shown. From Figure 14 It can be seen that when intervention was performed at 18h and 72h, the myocardial injury markers BNP and cTnT in the CLP group rats were significantly higher than those in the Sham group, while the myocardial injury markers BNP and cTnT in the WY group rats were significantly lower than those in the CLP group, suggesting that myocardial injury was reduced.

[0200] The comparison results of inflammatory factors (IL-1β, IL-6, IL-10, TNF-α) in each group of rats are as follows: Figure 15 As shown. From Figure 15 It can be seen that at the 18-hour time point, the levels of inflammatory factors IL-1β, IL-6, IL-10, and TNF-α in the CLP group rats were significantly higher than those in the Sham group, while the levels of inflammatory factors IL-1β, IL-6, and TNF-α in the WY group rats were significantly lower than those in the CLP group. At the 72-hour time point, the levels of inflammatory factors IL-1β and IL-6 in the CLP group rats were significantly higher than those in the Sham group, while the level of inflammatory factor IL-6 in the WY group rats was significantly lower than that in the CLP group, suggesting that the traditional Chinese medicine composition of the present invention has a good anti-inflammatory effect.

[0201] The comparison results of oxidative stress factors (MDA, GSH, SOD) in each group of rats are as follows: Figure 16 As shown. From Figure 16It can be seen that at the 18-hour time point, the oxidative stress factor MDA in the CLP group rats was significantly higher than that in the Sham group, while GSH and SOD were significantly lower; in the WY group rats, the oxidative stress factor MDA was significantly lower than that in the CLP group, while GSH was significantly higher. At the 72-hour time point, the oxidative stress factor MDA in the CLP group rats was significantly higher than that in the Sham group, while GSH was significantly lower; in the WY group rats, the oxidative stress factor MDA was significantly lower than that in the CLP group, while SOD was significantly higher. This suggests that the Wen Yang Jie Du formula can reduce the oxidative stress response in sepsis to a certain extent.

[0202] The comparison results of the expression of autophagy-related proteins (P62, LC3, Beclin) in each group of rats are as follows: Figure 17 As shown. From Figure 17 It can be seen that at the 18-hour intervention point, the levels of autophagy-related proteins P62 and LC3 in the CLP group rats were significantly lower than those in the Sham group, while the level of Beclin in the CLP group rats was significantly higher than that in the Sham group rats; the levels of autophagy-related proteins P62 and LC3 in the WY group rats were significantly higher than those in the CLP group rats. At the 72-hour intervention point, the level of autophagy-related protein Beclin in the CLP group rats was significantly higher than that in the Sham group rats, while the level of autophagy-related protein Beclin in the WY group rats was significantly lower than that in the CLP group rats. This suggests that in the early stage of sepsis, the Wen Yang Jie Du formula can effectively increase myocardial autophagy, and with the increase of intervention time, at 72 hours, the Wen Yang Jie Du formula can inhibit excessive myocardial autophagy.

[0203] Pathological sections of rat myocardium in each group are as follows: Figure 18 As shown. From Figure 18 It can be seen that, compared with the Sham group, the CIP group rats showed significant inflammation and myocardial edema; the WY group rats showed reduced inflammatory exudation and myocardial edema compared with the CLP group. This suggests that the warming and detoxifying formula of the present invention can effectively alleviate myocardial damage caused by sepsis.

[0204] The comparison results of autophagosomes in the myocardial endothelial cells of rats in each group are as follows: Figure 19 As shown. From Figure 19 It can be seen that, compared with the Sham group, autophagy in the myocardial endothelial cells of rats in the CIP group was significantly reduced; autophagy in the myocardial endothelial cells of rats in the WY group was increased compared with that in the CLP group. This suggests that the traditional Chinese medicine composition of the present invention can increase autophagy in myocardial endothelial cells, possibly by increasing beneficial autophagy to exert a protective effect on myocardial cells.

[0205] The above descriptions are merely some specific embodiments of the present invention. For those skilled in the art, various modifications and improvements can be made without departing from the inventive concept of the present invention, and all such modifications and improvements fall within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine composition for treating sepsis-induced cardiomyopathy due to yang deficiency and blood stasis, characterized in that, It is made from the following raw materials by weight: 10-50 parts ginseng, 5-20 parts rhubarb, 10-30 parts aconite, 5-20 parts coptis, 5-30 parts scutellaria, and 5-20 parts safflower.

2. The traditional Chinese medicine composition for treating sepsis-induced cardiomyopathy due to yang deficiency and blood stasis according to claim 1, characterized in that, It is made from the following raw materials by weight: 20 parts ginseng, 6 parts rhubarb, 15 parts aconite, 6 parts coptis, 6 parts scutellaria, and 10 parts safflower.

3. The traditional Chinese medicine composition for treating sepsis-induced cardiomyopathy due to yang deficiency and blood stasis according to claim 1 or 2, characterized in that, The aconite mentioned is processed aconite.

4. The traditional Chinese medicine composition for treating sepsis-induced cardiomyopathy due to yang deficiency and blood stasis according to claim 1 or 2, characterized in that, It also includes medically acceptable excipients.

5. The traditional Chinese medicine composition for treating sepsis-induced cardiomyopathy with yang deficiency and toxic stasis syndrome according to claim 1 or 2, characterized in that, The dosage form of the traditional Chinese medicine composition is decoction, granules, powder, tablets, pills or capsules.

6. The use of the traditional Chinese medicine composition for treating sepsis-induced cardiomyopathy with yang deficiency and toxic stasis syndrome as described in any one of claims 1 to 5 in the preparation of a medicament for treating sepsis-induced cardiomyopathy with yang deficiency and toxic stasis syndrome.

Images