CN114010653B

CN114010653B - Application of composition comprising ginsenoside Rg1 and fucoidin in preparation of medicine for reducing expression of tissue factor

Info

Publication number: CN114010653B
Application number: CN202111444654.XA
Authority: CN
Inventors: 刘文涛; 陈琪; 李岩; 姜春懿; 胡亮; 吴雪丰; 王玉; 林彤彤; 贾茹梦
Original assignee: Nanjing Medical University
Current assignee: Nanjing Medical University
Priority date: 2020-12-11
Filing date: 2020-12-11
Publication date: 2023-07-25
Anticipated expiration: 2040-12-11
Also published as: CN112546056B; CN114028416A; CN114010653A; CN112546056A; CN114159460A; CN114028416B; CN114159460B

Abstract

The invention provides a composition for treating chemotherapy-induced peripheral neuropathy and application thereof, and belongs to the field of medicines. The invention provides a composition for treating chemotherapy-induced peripheral neuropathy, which comprises ginsenoside Rg1 and fucoidin. The composition can obviously inhibit the expression of proinflammatory factors such as TNF-alpha, IL-1 beta, IL-6 and the like, thereby generating anti-inflammatory and pro-repairing effects on peripheral nerves and enhancing the resistance; in addition, the composition can also remarkably improve microcirculation disturbance, up-regulate SR-A expression, further enhance the phagocytosis and clearance capacity of macrophages on HMGB1, inhibit the overexpression of TF, relieve the microcirculation disturbance and the neuropathic pain caused by chemotherapeutics by multiple targets and multiple mechanisms, and fundamentally inhibit the development of CIPN, thereby achieving the aim of safe and effective treatment and providing a new treatment strategy for clinic.

Description

Application of composition comprising ginsenoside Rg1 and fucoidin in preparation of medicine for reducing expression of tissue factor

The application is a divisional application of 12/11 th year 2020, 202011457399.8 and entitled "composition for treating chemotherapy-induced peripheral neuropathy and application thereof".

Technical Field

The invention belongs to the field of medicines, and particularly relates to a composition for treating chemotherapy-induced peripheral neuropathy and application thereof.

Background

A tumor is a disease that is a serious hazard to human health and its treatment is very difficult. Although there are some targeted drug therapies, chemotherapy remains one of the most common approaches to treating malignant tumors. Unfortunately, chemotherapy often causes serious side effects such as pain, vomiting, dizziness, and alopecia. Especially pain, can cause the patient to become more anxious and even depressed, which can severely affect the patient's mental state and ultimately prevent the treatment of the tumor. Among the side effects of tumors, chemotherapy-induced peripheral neuropathy (CIPN) is a very painful response, mainly manifested by paresthesias, especially pain that produces cold tactile and mechanical hypersensitivity, and the sensation of this pain is sometimes so intense that the chemotherapeutic patient has to reduce the dose of chemotherapeutic drug to relieve the pain, severely hampering the therapeutic effect of the Chemotherapy. About 30% -40% of patients are reported to develop CIPN, so that the study of relevant mechanism and therapeutic measures of CIPN has important clinical value and social significance.

The existing medicines for researching and treating CIPN mainly protect the nerve of needles and relieve pain symptoms, and treat symptoms without treating the root cause, for example, neuroprotective medicines comprise glutathione, calcium and magnesium, vitamin B, vitamin E, acetyl l-carnitine and the like, so that nerve injury can be relieved, and the symptoms of paresthesia can be improved; neuropathic pain symptomatic treatment drugs include tricyclic antidepressants, 5-HT and norepinephrine reuptake inhibitors (SNRI), antiepileptics, etc., and adverse gastrointestinal reactions such as nausea, dry mouth, constipation, cardiovascular adverse reactions (such as postural hypotension), central nervous system toxicity (such as insomnia, anxiety, headache), etc. can occur after long-term use of such drugs. According to the american society of clinical oncology (ASCO 2020), due to the lack of high quality, consistent evidence, the use of any drug for the prevention of chemotherapy-induced peripheral neuropathy is not recommended, and for patients with pain due to CIPN, the clinician may use duloxetine treatment with moderate quality of evidence and moderate intensity of recommendation. Thus, to date, there has been no clear and effective method to prevent CIPN, reduce the dosage of chemotherapeutic agents, and terminate the use of chemotherapeutic agents, which is still the primary method of treating CIPN. Because of the complex mechanism of CIPN and the limitations of existing therapeutic agents, finding new therapeutic agents is a problem to be solved in the clinic.

Disclosure of Invention

In order to solve the above problems, the present invention provides a composition for treating chemotherapy-induced peripheral neuropathy and use thereof. The composition has good effect of treating CIPN, and the components of the composition are natural components, so that the composition is safe and has no toxic or side effect.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides a composition for treating chemotherapy-induced peripheral neuropathy, comprising ginsenoside Rg1 and fucoidin.

Preferably, the mass ratio of the ginsenoside Rg1 to the fucoidin is 1:1-1:5.

The invention provides application of the composition in the technical scheme in preparing medicines for treating chemotherapy-induced peripheral neuropathy.

The invention provides application of the composition in the technical scheme in preparing medicines for improving microcirculation disturbance.

The invention provides application of the composition in the technical scheme in preparing medicines for reducing the expression of tissue factors.

The invention provides application of the composition in the technical scheme in preparing a medicament for inhibiting the expression of sciatic nerve inflammatory factors of peripheral neuropathy induced by chemotherapy.

The invention provides application of the composition in the technical scheme in preparing a medicament for reducing the expression of high mobility group protein B1.

The invention provides application of the composition in the technical scheme in preparing a medicine for increasing the mechanical pain threshold of chemotherapy-induced peripheral neuropathy.

The beneficial effects are that:

the invention provides a composition for treating chemotherapy-induced peripheral neuropathy, comprising ginsenoside Rg1 and fucoidin. The composition can obviously inhibit the expression of proinflammatory factors such as TNF-alpha, IL-1 beta, IL-6 and the like, thereby generating anti-inflammatory and pro-repairing effects on peripheral nerves and enhancing the resistance; in addition, the composition can also remarkably improve microcirculation disturbance, enhance the phagocytosis and clearance capacity of macrophages to HMGB1 (high mobility group protein B1) through SR-A mediated signal paths, inhibit the overexpression of TF (tissue factor), relieve the microcirculation disturbance and neuropathic pain caused by chemotherapeutics through multiple targets and multiple mechanisms, and fundamentally inhibit the development of CIPN, thereby achieving the aim of safe and effective treatment and providing a new treatment strategy for clinic. The results of the examples show that: the composition provided by the invention can obviously improve the mechanical pain threshold of the CIPN model mouse induced by oxaliplatin, obviously improve the microcirculation disturbance condition of the CIPN model mouse, and has obvious effect on the treatment of CIPN.

Drawings

FIG. 1 is a graph showing the effect of each treatment group on the plantar pain threshold of mice in example 3;

FIG. 2 is a graph showing the effect of each treatment group on mice plantar microcirculation in example 3, wherein A is a laser Doppler blood flow graph and B is a laser Doppler blood flow graph statistical graph;

FIG. 3 is a graph showing the effect of each treatment group on inflammatory factors in mice in example 3, wherein A is the effect of each treatment group on IL-1β mRNA levels; b is the effect of each treatment group on IL-6IL-6mRNA levels; c is the effect of each treatment group on TNF- α mRNA levels; d is a data statistical graph of HMGB1 expression conditions in the plasma of each treatment group; # p <0.01, # p <0.001, indicating significant differences compared to the control group; * p <0.05, < p <0.01, < p <0.001, indicating a significant difference compared to the oxaliplatin group;

FIG. 4 effect of each treatment group on the pain threshold and plantar blood flow in mice in example 4, wherein A is the effect of each treatment group on the pain threshold in mice plantar; b is the influence of each treatment group on the plantar microcirculation of the mice; * p <0.05, < p <0.01, < p <0.001, indicating significant differences compared to the control group; #p <0.05, #p <0.01, #p <0.001, indicating significant differences compared to oxaliplatin group;

FIG. 5 effect of each treatment group on HMGB1 and TF in mouse plasma in example 4, wherein A is the effect of each treatment group on HMGB1 in mouse plasma; b is the effect of each treatment group on TF in mouse plasma; * p <0.05, < p <0.01, < p <0.001, indicating significant differences compared to the control group; #p <0.05, #p <0.01, #p <0.001, indicating significant differences compared to oxaliplatin group; and p <0.05, & p <0.01, indicating a significant difference compared to the oxaliplatin + fucoidan group.

Detailed Description

The invention provides a composition for treating chemotherapy-induced peripheral neuropathy, comprising ginsenoside Rg1 and fucoidin. The invention has no special requirements on the sources of ginsenoside Rg1 and fucoidin, and can be obtained by adopting common commercial products in the field. In the invention, ginsenoside Rg1 and fucoidan are purchased from sigma company, and the mass ratio of the ginsenoside Rg1 to the fucoidan is preferably 1:1-1:5. The ginsenoside Rg1 and fucoidin compound belongs to a natural compound product, and has the advantages of low toxicity, wide application, simple preparation, safety and effectiveness; the application of the compound in the CIPN mouse model not only can eliminate the nerve inflammation, but also can improve the plantar blood flow and relieve microcirculation disturbance, and can promote the phagocytosis and removal capacity of macrophages on harmful substances in the organism, so that the CIPN can be effectively relieved in multiple aspects, a strong superposition effect is generated, and a novel effective treatment means is provided for the clinical problem to be solved urgently, such as neuralgia induced by chemotherapy, and the patients suffering from the neuralgia.

The invention provides application of the composition in the technical scheme in preparing medicines for treating chemotherapy-induced peripheral neuropathy. The composition provided by the invention can obviously improve the mechanical pain threshold of a CIPN model mouse induced by chemotherapy, obviously improve the microcirculation disturbance condition of the CIPN model mouse, and has obvious effect on the treatment of CIPN.

The invention provides application of the composition in the technical scheme in preparing medicines for improving microcirculation disturbance. The composition provided by the invention can significantly improve the circulation disorder of CIPN induced by chemotherapy, and has a significant effect on the treatment of CIPN.

The invention provides application of the composition in the technical scheme in preparing a medicament for reducing tissue factor expression. The composition provided by the invention can obviously reduce the expression of Tissue Factor (TF), and has obvious effect on the treatment of CIPN.

The invention provides application of the composition in the technical scheme in preparing a medicament for inhibiting the expression of sciatic nerve inflammatory factors of peripheral neuropathy induced by chemotherapy. The composition provided by the invention can obviously inhibit the accumulation of ischial nerve inflammatory factors TNF-alpha, IL-1 beta and IL-6, and has obvious effect on the treatment of CIPN.

The invention provides application of the composition in the technical scheme in preparing a medicament for reducing the expression of high mobility group box protein B1 (HMGB 1). The composition provided by the invention can obviously reduce the expression of HMGB1 and has obvious effect on the treatment of CIPN.

The invention provides application of the composition in the technical scheme in preparing a medicine for increasing the mechanical pain threshold of chemotherapy-induced peripheral neuropathy. The composition can obviously raise the mechanical pain threshold of peripheral neuropathy induced by chemotherapy, relieve the nerve pain of CIPN, and has obvious effect on the treatment of CIPN.

The auxiliary materials and the dosage forms of the medicines in the technical scheme have no special requirements, and the auxiliary materials and the dosage forms which are conventional in the field are adopted; or directly using the mixture of ginsenoside Rg1 and fucoidin as therapeutic drug without adding adjuvants.

To further illustrate the present invention, the compositions provided herein for treating chemotherapy-induced peripheral neuropathy and the use thereof are described in detail below in conjunction with the examples, but are not to be construed as limiting the scope of the present invention.

Example 1

A composition for treating chemotherapy-induced peripheral neuropathy comprises ginsenoside Rg1 and fucoidin, wherein the purity of ginsenoside Rg1 is not less than 95%, the purity of fucoidin is not less than 95%, and no special requirements on granule and diameter are provided. The mass ratio of the ginsenoside Rg1 to the fucoidin is 1:1.

the preparation method comprises the following steps: 40mg of ginsenoside Rg1 powder and 40mg of fucoidin powder are dissolved in 5ml of sterile water, vortex is vibrated to mix and dissolve completely.

Example 2

A composition for treating chemotherapy-induced peripheral neuropathy comprises ginsenoside Rg1 and fucoidin, wherein the purity of ginsenoside Rg1 is not less than 95%, the purity of fucoidin is not less than 95%, and no special requirements on granule and diameter are provided. The mass ratio of the ginsenoside Rg1 to the fucoidin is 1:5.

the preparation method comprises the following steps: 40mg of ginsenoside Rg1 powder and 200mg of fucoidin powder are dissolved in 5ml of sterile water, vortex is vibrated to mix and dissolve completely.

Example 3

A composition for treating chemotherapy-induced peripheral neuropathy comprises ginsenoside Rg1 and fucoidin, wherein the purity of ginsenoside Rg1 is not less than 95%, the purity of fucoidin is not less than 95%, and no special requirements on granule and diameter are provided. The mass ratio of the ginsenoside Rg1 to the fucoidin is 1:2.5.

the preparation method comprises the following steps: 40mg of ginsenoside Rg1 powder and 100mg of fucoidin powder are dissolved in 5ml of sterile water, vortex is vibrated to mix and dissolve completely.

Evaluation of the Effect of the inventive composition on treatment of CIPN

Experimental protocol:

1 experimental animal: male C57BL/6J mice and male ICR mice, SPF grade, day age 6-8 weeks, supplied by Shanghai national institute of family planning science laboratory animal manager, SR-AKO mice were given benefit by the teaching of university Chen Qi of Nanjing medical science and were bred in the university of Nanjing medical science laboratory animal center. The experiments were carried out using ICR mice, C57BL/6J mice, SR-AKO mice. All experimental animals are fed in independent environments with alternating brightness for 12 hours, drinking water and ingestion are free, the temperature of the feeding environment is maintained at 22+/-2 ℃, and the experiment is carried out after the experimental animals are adapted to the environment for 1 week. All experimental procedures followed the international committee on pain research association ethics regulations.

2, experimental method:

(1) CIPN model: SPF-grade adult healthy male ICR mice are selected, and the test mice are randomly divided into 7 groups (n=6), namely a control group, an oxaliplatin group (model group), an oxaliplatin+fucoidan group, an oxaliplatin+ginsenoside Rg1 group, an oxaliplatin+fucoidan+ginsenoside Rg1 (1:2.5) group, an oxaliplatin+fucoidan+ginsenoside Rg1 (1:1) group, and an oxaliplatin+fucoidan+ginsenoside Rg1 (1:5) group.

Wherein the oxaliplatin+fucoidan group starts to administer fucoidan at a concentration of 200mg/kg 7 days before molding, the oxaliplatin+ginsenoside Rg1 group starts to administer ginsenoside Rg1 at a concentration of 80mg/kg 7 days before molding, the control group starts to administer physiological saline at a concentration of 0.1ml/10g 7 days before molding, the oxaliplatin+fucoidan+ginsenoside Rg1 (1:2.5) group starts to administer 10ml/kg of the composition of example 3 to mice 7 days before molding, the oxaliplatin+fucoidan+ginsenoside Rg1 (1:1) group starts to administer 10ml/kg of the composition of example 1 to mice at 7 days before molding, the oxaliplatin+fucoidan+ginsenoside Rg1 (1:5) group starts to administer 10ml/kg of the composition of example 2 to mice at 7 days before molding, and five groups after molding start to administer the composition of oxaliplatin+fucoidan+ginsenoside Rg1 (1:1) respectively, and the injection of oxaliplatin+fucoidan+ginsenoside Rg1 (1:1) is continued for a period of time of 14 days after molding, and the injection of oxaliplatin is continued for the mice to continue to administer the treatment group to the animal model for 14 days. Behavioural measurements were performed daily from the mouse molding, and plantar Doppler blood flow measurements were performed at 14d after molding to examine the microcirculation disturbance.

(2) Mechanical foot-retraction reflectance threshold measurement: the behavioural assay of the present invention, i.e. the mechanical pain threshold measurement of mice, is von freyhairst (fibrotic pain tester), reference is: gu H, wang C, li J, yang Y, sun W, jiang C, li Y, ni M, liu WT, cheng Z, hu L.high mobility group box-1-toll-like receptor 4-phosphotidylinosol 3-kinase/protein kinase B-mediated generation of matrix metalloproteinase-9in the dorsal root ganglion promotes chemotherapy-induced peripheral neuroplath.int J cancer.2020May15;146 (10) 2810-2821.Doi:10.1002/ijc.32652.Epub 2019Sep 14.PMID:31465111. The cellosilk pain measuring instrument consists of 20 Von Frey hair cilia mechanical stimulation needles, is mainly used for evaluating the touch feeling of skin, and can provide the Von Frey hair stimulation force of 0.008 g-300 g. The thickness and the extension of Von Frey hair nylon filaments determine the amount of stimulation provided. During experiments, a Von Frey hair cilia mechanical stimulation needle with proper thickness is selected according to actual conditions, skin is stimulated vertically, and the stimulation force can be adjusted by adjusting and replacing the Von Frey hair cilia mechanical stimulation needle until the Von Frey hair cilia mechanical stimulation needle is bent. When the paw of a rodent is subjected to a mechanical stimulus, there is a retractive reflex, which can be applied to the sole of a rat or mouse.

The invention performs pain threshold measurement of the right paw of the mouse daily according to the method described above. In a quiet environment, mice were placed in a clear compartment over a wire mesh, and allowed to freely move for 20min. After the exploring behavior of the mice stops and limbs are contacted with the wire gauze, the skin of the bottoms of feet of the mice are perpendicularly stimulated by Von-Fery fibers with different folding forces, the mice are gradually pressurized to slightly bend the filaments, the 4s is maintained, whether the mice have foot contraction reaction is observed, the actions of foot contraction, licking and the like of the mice are regarded as positive reactions, and the maximum pressure causing the foot contraction reaction of the mice is recorded as a mechanical stimulation foot contraction reflection threshold value. At each measurement interval of 3min, the mechanical stimulus foot-reduction reflection threshold of 50% is calculated by adopting a sequential method, the average value is obtained after 3 times of measurement, and the detection result is shown in figure 1.

(3) Doppler blood flow measurement: the flow velocity and the flow quantity of blood are measured by an ultrasonic Doppler blood flow instrument, an ultrasonic probe with a fixed position emits ultrasonic waves, the ultrasonic waves are received by erythrocytes in the blood, then the erythrocytes are used as a wave source, the ultrasonic probe receives reflected waves of the erythrocytes, and the flow velocity of the blood can be calculated according to a Doppler effect formula by utilizing the frequency difference between the emitted waves and the reflected waves of the ultrasonic waves. The invention removes hair from the lower body of the mouse, carries out Doppler blood flow measurement, mainly observes the hind paws of the mouse, monitors the microcirculation disturbance condition of the mouse, and under the same contrast, the darker the red color is, the richer the blood flow is, the darker the blue color is, the condition that the microcirculation disturbance condition is a certain is represented, the blood flow is blocked, and the detection result is shown in figure 2.

(4) The real-time quantitative polymerase chain reaction (qPCR) detects the mRNA levels of IL-1β, IL-6 and TNF- α.

A. Extracting RNA: after the cell administration treatment, RNA was extracted with Trizol reagent according to the operation requirements, and the concentration was measured.

B. Reverse transcription reaction: the process is carried out by using a HiScript II Q RT SuperMix kit, and the reaction system is as follows: mu.l of 5X HiScript II Q RT SuperMix,500ng of template RNA and finally adding RNase Free ddH20 to make up the volume to 10. Mu.l of the total reaction system; the PCR instrument sets the reaction program, and the cDNA sample is obtained by reverse transcription at 50 ℃,15 min-85 ℃ for 5s.

Qpcr reaction: the cDNA sample was diluted 5-fold, and the reaction system was: mu.l of the upstream primer, 2. Mu.l of the downstream primer, 1. Mu.l of the diluted cDNA template and 5. Mu.l of 2X SYBR qPCR Master Mix were each added to an octant so that the total reaction volume was 10. Mu.l, and the mixture was vortexed and homogenized, and the primers were as shown in Table 1.

The reaction conditions were as follows: 95 ℃ for 30s;40 cycles, 95 ℃,10sec;60 ℃ for 30s. The dissolution curve is 95 ℃ for 15s;60 ℃ for 60s;95℃for 15s.

The Ct value obtained is analyzed as a result, and the calculation formula is as follows: 2- ΔΔct, Δct=ct gene-CtACTIN, delta CT = delta CT-control group delta CT mean, the relative quantification method is adopted to compare with the reference ACTIN.

TABLE 1 statistics of primer sequences

(5) Westernblot detects protein expression levels of HMGB 1.

(1) Animal sample preparation: after 7 days of mouse CIPN modeling, mouse plasma was taken and 190. Mu.l RIPA lysate containing the phosphatase inhibitor and the protease inhibitor PMSF was added to each sample. Adding 10ul of plasma sample, standing on ice for 30min, measuring protein concentration in tissue sample by BCA method, adding 5×loading buffer proportionally, mixing by vortex, boiling for 5min to denature protein; samples were stored at-20 ℃.

(2) Electrophoresis and transfer: taking 30 mug plasma sample of total protein, performing SDS-PAGE electrophoresis with 8%,10% or 15% gel, stopping electrophoresis when protein separation is proper; and transferring the cut adhesive onto a PVDF film, and transferring the film at a constant current of 300mA, wherein the film transferring time is determined by the molecular weight of the target protein.

(3) Blocking and antibody incubation: 10% whole milk powder (TBST dissolved) or 5% BSA+5% milk powder solution was blocked for 2h at room temperature; according to the following steps of 1: the betA-Actin antibody and the HMGB1 antibody are diluted in a proportion of 1000, and the strips are placed in primary anti-dilution liquid and incubated for 16h at 4 ℃. After incubation, the membrane was washed 3 times with 10min each time with TBST. Preparing a secondary antibody solution according to a dilution ratio of 1:5000, and incubating the secondary antibodies corresponding to the primary antibodies of the strips with a shaking table at room temperature for 2 hours, wherein after incubation, TBST washes the membrane for 4 times, each time for 10 minutes.

(4) Color development: developing solution is prepared from solution A and solution B in the ECL chemiluminescence kit according to the proportion of 1:1, the strips are placed on an exposure table of a gel imager, the developing solution is evenly paved on the surface of the film, the film is incubated for 1-2min, and a set program is operated to perform exposure imaging.

(5) Data analysis: the gray scale of each group of data images was scanned using Gel Pro or Image J software, and data statistics and analysis were performed using EXCEL, the results of which are shown in FIG. 3.

FIG. 1 is the effect of each treatment group on the pain threshold of mice. As can be seen from the results of fig. 1, the combination of ginsenoside Rg1 and fucoidin can significantly raise the mechanical pain threshold of mice, and the combined administration gradually shows advantages over the single administration of both at the beginning of 10 th day, lasting for 21 days, and the effect of alleviating the mechanical pain threshold of mice by the combined administration is significantly better than that by the single administration of both. In 2017, related literature reports that CIPN models were established using oxaliplatin, treated with duloxetine at three concentrations of 10, 30, 60mg/kg, and measured for mechanical pain threshold in mice using vonFrey filament tests. The skin of the hind paw of the mouse was stimulated 10 times with a bending force of 0.4g, and the mice were scored positive for a total of 100%. The results showed that 10mg/kg duloxetine had no significant effect on alleviating mechanical pain in mice, a dose of 30mg/kg could alleviate pain in about 20% of mice, and a dose of 60mg/kg could improve pain in about 40% of mice, but this dose could trigger a partial mouse somnolence (literature: kim W, chung Y, choi S, min BI, kim SK.Duloxetine Protects against Oxaliplatin-Induced Neuropathic Pain and Spinal Neuron Hyperexcitability in Rodens.int J Mol Sci.2017Dec 5;18 (12): 2626.Doi:10.3390/ijms18122626.PMID:29206213; PMCID: PMC 5751229.) with serious adverse effects). The composition of the invention can obviously improve the mechanical pain threshold of mice, and has no adverse side effect.

Fig. 2 shows the effect of each treatment group on the plantar microcirculation of mice, and it is clear from the results of fig. 2 that the mice given with oxaliplatin (L-OHP) are serious in microcirculation disturbance, and the blood flow condition of the mice given with ginsenoside Rg1 alone is not significantly improved, but the mice given with fucoidan are significantly improved in blood flow condition, and the mice given with fucoidan are combined to further improve microcirculation disturbance, wherein the effects of oxaliplatin+fucoidan+ginsenoside Rg1 (1:2.5) are most remarkable.

Fig. 3 shows the effect of each treatment group on inflammatory factors in mice, and mainly considers that the combination administration of fucoidan and ginsenoside can significantly inhibit the expression of inflammatory factors and the accumulation of HMGB 1. Wherein A is the effect of each treatment group on IL-1β mRNA levels; b is the effect of each treatment group on IL-6mRNA levels; c is the effect of each treatment group on TNF- α mRNA levels; d is a statistical plot of HMGB1 expression in plasma of each treatment group. As can be seen from fig. 3, the combined administration of ginsenoside and fucoidin can significantly reduce the expression levels of inflammatory factors IL-1 beta, TNF-alpha and IL-6 in the blood of mice, and reduce the excessive accumulation of HMGB1, wherein the oxaliplatin+fucoidin+ginsenoside Rg1 (1:2.5) group has the most significant effect, and can significantly alleviate CIPN.

Example 4

Test protocol: the test was divided into 5 treatment groups, which were control group, oxaliplatin group (model group), oxaliplatin+fucan group, SR-a knockout+oxaliplatin group, SR-a knockout+oxaliplatin+fucan group, and n=6 above, wherein the control group, oxaliplatin group, and oxaliplatin+fucan group were C57BL/6J mice, SR-a knockout+oxaliplatin group, SR-a knockout+oxaliplatin+fucan group were SA-A knockout mice, oxaliplatin+fucan group, and +sr-a knockout+oxaliplatin+fucan group were started to administer mouse fucan (200 mg/kg) to the celiac cavity of the mouse in advance for 7 days, and the oxaliplatin group, oxaliplatin+fucan group, SR-a knockout+oxaliplatin group, and SR-a knockout+fucan group were injected into the oxaliplatin+fucan group mice (3 mg/kg) on day 8, and the treatment was continuously performed for the animals until the treatment of the group was stopped for 14 days. The behavioral measurement was performed daily from the molding of mice, and the plantar Doppler blood flow measurement was performed at 14d after the molding, and the microcirculation disturbance condition and the expression level of each factor were examined by Westernblot, and the results are shown in FIGS. 4 to 5.

FIG. 4 shows the effect of each treatment group on the pain threshold and microcirculation disturbance of mice in vivo test, wherein A is the effect of each treatment group on the pain threshold of mice, and B is the effect of each treatment group on the circulatory disturbance. From the results of fig. 4, it can be seen that fucan can significantly reduce the pain threshold of oxaliplatin+fucan group and promote the microcirculation of mice, but the SR-a knockout+oxaliplatin+fucan group is not greatly improved, which means that the pain relieving effect and the microcirculation relieving effect of fucan on CIPN mice depend on SR-a receptors, and SR-a-/-completely eliminates the pain relieving effect and the microcirculation relieving effect of fucan, namely, the pain relieving mechanism of fucan in chemotherapy is the effect of SR-a mediated pain relieving effect of fucan and the effect of increasing blood perfusion of feet of mice.

FIG. 5 shows the effect of treatment groups on HMGB1 and TF expression in vivo experiments, wherein A is the effect of each treatment group on HMGB1 expression and B is the effect of each treatment group on TF expression. From the results of fig. 5, it can be seen that fucan significantly reduced HMGB1 and TF in the oxaliplatin+fucan group, but did not significantly improve the SR-a knockout+oxaliplatin+fucan group, indicating that fucan inhibited HMGB1 and TF in CIPN mice is dependent on SR-a receptor, and that SR-a-/-completely abolished the inhibition of HMGB1 and TF by fucan, i.e., fucan alleviating mechanism of chemotherapeutic pain was that fucan reduced accumulation of HMGB1 and TF expression in plasma of mice through SR-a signaling pathway.

The results of the above examples show that the composition of ginsenoside Rg1 and fucoidin provided by the invention can significantly improve the mechanical pain threshold of the CIPN model mouse induced by oxaliplatin, significantly improve the microcirculation disturbance condition of the CIPN model mouse, has significant effects on CIPN treatment, is safe and reliable, has low toxicity and wide easily-accepted population, and provides a new treatment strategy for clinic.

Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.

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Claims

1. Application of a composition comprising ginsenoside Rg1 and fucoidin or a composition comprising ginsenoside Rg1 and fucoidin in preparing a medicament for relieving peripheral nerve pain caused by oxaliplatin;

the mass ratio of the ginsenoside Rg1 to the fucoidin is 1:1-1:5;

the medicine is a medicine for reducing tissue factor expression.