CN114028455A

CN114028455A - Preparation process of honeysuckle detoxification mixture capable of improving immunity

Info

Publication number: CN114028455A
Application number: CN202111558988.XA
Authority: CN
Inventors: 唐强; 李书霖; 王有鹏
Original assignee: Second Affiliated Hospital Of Heilongjiang University Of Traditional Chinese Medicine
Current assignee: Second Affiliated Hospital Of Heilongjiang University Of Traditional Chinese Medicine
Priority date: 2021-12-20
Filing date: 2021-12-20
Publication date: 2022-02-11

Abstract

The invention relates to a preparation process of a honeysuckle detoxification mixture capable of improving immunity, which is prepared from the following raw material medicaments in percentage by weight (in terms of prepared 1000ml of medicaments): 18.75 percent of honeysuckle, 18.75 percent of forsythia, 18.75 percent of atractylodes, 12.5 percent of platycodon root, 12.5 percent of mulberry leaf, 12.5 percent of eupatorium and 6.25 percent of liquorice; soaking the above seven materials in water, decocting, standing, filtering, concentrating the filtrate under reduced pressure to 1000ml, and packaging. The preparation process of the honeysuckle detoxification mixture capable of improving the immunity improves the clinical decoction into a mixture convenient to take, carry and store, has the effects of clearing away heat and toxic materials, strengthening the spleen to eliminate dampness and benefiting the lung and relieving sore throat, and is used for patients with fever and headache, cough with excessive phlegm, sore throat, short breath and frequent sweating caused by warm diseases of wind heat, damp heat and the like.

Description

Preparation process of honeysuckle detoxification mixture capable of improving immunity

[ technical field ] A method for producing a semiconductor device

The invention relates to a preparation method of a traditional Chinese medicine mixture, in particular to a preparation process of a honeysuckle detoxification mixture capable of improving immunity, and belongs to the technical field of traditional Chinese medicines.

[ background of the invention ]

The novel coronavirus pneumonia belongs to the categories of epidemic diseases and wet plagues in traditional Chinese medicine, the etiology attribute of the novel coronavirus pneumonia is pathogenic factors of the wet toxin, and the pathogenesis characteristic of the novel coronavirus pneumonia is dampness pathogen stasis. The disease is divided into the following four types according to the transmission rule of the disease: syndrome of dampness-warm stagnating in lung, syndrome of phlegm-heat obstructing in lung, syndrome of pathogen-toxin blocking in lung, syndrome of pathogen-toxin mongolian orifices. The clinical manifestations are as follows: fever and cough are not the only first symptoms of the new coronary pneumonia, and digestive system symptoms, nervous system symptoms, cardiovascular system symptoms, ophthalmic symptoms, or only mild muscle soreness of limbs or waist and back and the like can be the first symptoms of the new coronary pneumonia. Some patients have unobvious symptoms, but have strong infectivity, and only show low fever, slight hypodynamia and the like, and have no pulmonary inflammation, and the patients mostly recover after 1 week. Severe cases often develop dyspnea after 1 week, and severe cases rapidly progress to acute respiratory distress syndrome, septic shock, uncorrectable metabolic acidosis, and hemorrhagic coagulation dysfunction. The prognosis of most patients is good, the symptoms of children cases are relatively mild, the disease of a few patients is critical and even death occurs, and death cases are mostly seen in the elderly and patients with chronic basic diseases.

In traditional Chinese medicine, pathogenic factors of pestilence enter from mouth and nose, or enter membrane internally, and spread externally and internally, and rapidly. In the interior Jing, the cloud indicates that the healthy qi is in the interior and the pathogenic factors are not always present. Therefore, when the conventional prevention method is used, the traditional Chinese medicine intervention is carried out at an early stage, and the Chinese medicine formula of tonifying qi, strengthening exterior, clearing pestilence and detoxifying is orally taken in advance, so that the invasion and transmission of pathogenic factors of pestilence can be avoided, and the effects of preventing diseases before the diseases are prevented and preventing the diseases from being weakened are achieved.

At present, the novel coronavirus pneumonia is still in continuous epidemic, and the traditional Chinese medicine has unique effect on preventing the novel coronavirus pneumonia, and has larger social demand. Therefore, a preparation process of the honeysuckle detoxification mixture capable of improving the immunity is necessary.

[ summary of the invention ]

The invention aims to provide a preparation process of a honeysuckle detoxification mixture capable of improving immunity, which has the effects of clearing away heat and toxic materials, strengthening spleen and eliminating dampness, and benefiting lung and throat.

In order to achieve the purpose, the invention adopts the technical scheme that: a preparation process of a honeysuckle detoxification mixture capable of improving immunity comprises the following raw material medicaments in percentage by weight (based on the prepared 1000ml of medicament): 18.75 percent of honeysuckle, 18.75 percent of forsythia, 18.75 percent of atractylodes, 12.5 percent of platycodon root, 12.5 percent of mulberry leaf, 12.5 percent of eupatorium and 6.25 percent of liquorice; soaking the above seven materials in water, decocting, standing, filtering, concentrating the filtrate under reduced pressure to 1000ml, and packaging.

The preparation process of the honeysuckle detoxification mixture capable of improving the immunity further comprises the following steps: the specific weight of the raw material medicaments is as follows: 37.5g of honeysuckle, 37.5g of fructus forsythiae, 37.5g of bighead atractylodes rhizome, 25g of platycodon grandiflorum, 25g of mulberry leaf, 25g of eupatorium and 12.5g of liquorice.

The preparation process of the honeysuckle detoxification mixture capable of improving the immunity further comprises the following steps: the soaking time in water is 1 hour, the decocting time is 40 minutes, and the net standing time is 24 hours.

The preparation process of the honeysuckle detoxification mixture capable of improving the immunity comprises the following steps: and adding sodium benzoate into the concentrated solution, and uniformly mixing.

Compared with the prior art, the invention has the following beneficial effects: the preparation process of the honeysuckle detoxification mixture capable of improving the immunity improves the clinical decoction into the mixture convenient to take, carry and store, has the effects of clearing away heat and toxic materials, strengthening the spleen to eliminate dampness and benefiting the lung and relieving sore throat, is used for patients with fever and headache, cough with excessive phlegm, sore throat, short breath and frequent sweating caused by warm diseases of wind heat, damp heat and has unique effect on preventing novel coronavirus pneumonia.

[ description of the drawings ]

FIG. 1 is a histopathological diagram of male rats at the end of the administration period of the honeysuckle detoxification mixture.

FIG. 2 is a histopathological diagram of female rats at the end of the administration period of the honeysuckle detoxification mixture.

FIG. 3 is a histopathological diagram of male rats at the end of recovery period of the antidotal mistura of Lianhua.

FIG. 4 is a histopathological view of female rats at the end of recovery period of the antidote mistura herb-withdrawal period.

[ detailed description ] embodiments

The invention relates to a honeysuckle detoxification mixture capable of improving immunity, which is prepared from the following raw material medicaments in percentage by weight (in a prepared 1000ml dosage): 18.75 percent of honeysuckle, 18.75 percent of forsythia, 18.75 percent of atractylodes, 12.5 percent of platycodon root, 12.5 percent of mulberry leaf, 12.5 percent of eupatorium and 6.25 percent of liquorice. In the present embodiment, the specific weights of the raw material drugs are: 37.5g of honeysuckle, 37.5g of fructus forsythiae, 37.5g of bighead atractylodes rhizome, 25g of platycodon grandiflorum, 25g of mulberry leaf, 25g of eupatorium and 12.5g of liquorice.

The preparation process of the honeysuckle detoxification mixture capable of improving the immunity comprises the following steps: soaking the above seven materials in water, decocting, standing, filtering, concentrating the filtrate under reduced pressure to 1000ml, and packaging.

Specifically, the water is added for soaking for 1 hour, the decoction time is 40 minutes, and the net standing time is 24 hours.

Further, sodium benzoate as a preservative was added to 1000ml of the concentrate, and mixed well.

The pharmacodynamics demonstration of the honeysuckle detoxification mixture prepared by the invention is as follows.

The antidotal mixture of the honeysuckle and the honeysuckle has the antipyretic effect: yeast induced fever test in rats

1.1 materials and instruments

1.1.1 test animals

SPF-grade Wistar rats, half male and female, initial body mass (180 + -20) g, purchased from Beijing Wintolite laboratory animal technologies, Inc., certification number: SCXK (Kyoto) 2016-. A breeding environment: the temperature is 23-25 ℃, the humidity is 55% -65%, and the artificial illumination is carried out for 12 h: alternating light and shade for 12 h. All groups of rats and mice are raised in standard cages and are fed with water freely. The treatment of animals in the experimental process conforms to the guidance comments on the animals being studied in good care issued by the national department of science and technology in 2006.

1.1.2 drugs and reagents

Honeysuckle detoxification mixture: the mixture is equivalent to 0.5g of crude drug per 1.0 ml. Storage conditions are as follows: refrigerating at 4 deg.C, storing under sealed condition, and re-heating to 37 deg.C for use in water bath during experiment; aspirin effervescent tablets (Aslick, batch No. 1905213); high-activity dry yeast (Angel Yeast Co., Ltd., lot: 20200418).

1.1.3 instruments and devices

FA2004 analytical balance (SOPTOP/shun constant-level instrument); an electric constant temperature water bath (Tianjin Tiantai instruments ltd); YP5002 electronic balance (positron instruments ltd, city, changzhou); LY-301C electronic thermometer (Beijing Lvyuan Hengtai medical equipment Co., Ltd.)

1.2 test methods

Rats (180 g and 200g, half male and female) are adapted to the environment for 3 days, from the 4 th day, the adaptive anal temperature measurement operation is carried out on the rats in the morning and at the evening for 2 times every day, vaseline is smeared around the anus of the rats before measurement, a thermometer is inserted into the anus of the rats after the rats empty excrement, the insertion depth of each time is kept consistent, the measurement is continuously carried out for 3 days and 2 times/day, and animals with the temperature fluctuation of not more than 0.5 ℃ and the average body temperature of 36.6-38.3 ℃ are selected as experimental objects. Fasting is not forbidden for 12h before the formal experiment, the body temperature is measured for 1 time every 1h in the morning of the experiment, the temperature is measured for 2 times continuously, and the average value is taken as the basic body temperature. Each of the male and female rats was randomly divided into 5 rats as a blank control group, and the other groups except the blank control group were injected with 10ml/kg of 20% dry yeast suspension subcutaneously in the back, and the blank control group was injected with an equal volume of physiological saline subcutaneously in the back. After 6 hours of heating, 50 rats (each half of male and female) with the body temperature raised by more than 0.8 ℃ are selected, and the male and female animals are randomly divided into a honeysuckle detoxification mixture low, medium and high dose group (1.8, 3.6 and 7.2g/kg), a model group and an aspirin group (a positive medicine group, 0.2g/kg), and each group comprises 10 rats (each half of male and female). The corresponding drugs are administered to each administration group by intragastric administration, and equal volume of distilled water is administered to the blank group and the model group by intragastric administration. The body temperature of rats in each group was measured and recorded at 1h, 2h, 3h, 4h and 5h after administration. The temperature rise Δ T of each group of rats at each time point was calculated.

Delta T-body temperature at fever-basal body temperature

1.3 test results

The results are detailed in table 1.

TABLE 1 Effect of antidotal mixture of Lonicera Japonica flos on fever of rats caused by Yeast

Note: p < 0.01 compared to blank; compared with model group^#P＜0.05，^##P＜0.01

The analysis in the table 1 shows that compared with the blank group, the body temperature of the rats in the model group is remarkably increased (P is less than 0.01) after the rats in the model group are subjected to pyrogenicity for 6 hours and other administration groups are subjected to administration for 1-5 hours, which indicates that the establishment of the fever model of the rats is successful. Compared with the model group, the body temperatures of rats in the aspirin group and the honeysuckle detoxification mixture high-dose group are respectively and remarkably reduced within 1-5h after administration (P < 0.05 or P < 0.01), the body temperatures of rats in the honeysuckle detoxification mixture medium-dose group are respectively and remarkably reduced within 2-5h after administration (P < 0.05 or P < 0.01), and the body temperatures of rats in the honeysuckle detoxification mixture low-dose group are remarkably reduced within 2h after administration (P < 0.05).

(II) the antidotal mixture of the honeysuckle has the anti-inflammatory effect: xylene induced mouse ear swelling test

2.1 materials and instruments

2.1.1 test animals

SPF grade ICR mice, male and female half, body mass (20 + -2) g, purchased from Liaoning Experimental animals technical GmbH, quality certification number: SCXK (Liao) 2020 and 0001. A breeding environment: the temperature is 23-25 ℃, the humidity is 55% -65%, and the artificial illumination is carried out for 12 h: alternating light and shade for 12 h. All groups of rats and mice are raised in standard cages and are fed with water freely. The treatment of animals in the experimental process conforms to the guidance comments on the animals being studied in good care issued by the national department of science and technology in 2006.

2.1.2 drugs and reagents

The content of the mixture is 0.5g per 1.0 ml. Storage conditions are as follows: refrigerating at 4 deg.C, storing under sealed condition, and re-heating to 37 deg.C for use in water bath during experiment; aspirin effervescent tablets (Aslick, batch No. 1905213); xylene (Fuyu Fine chemical Co., Ltd., Tianjin, lot number: 20190505);

2.1.3 instruments and apparatus

FA2004 analytical balance (SOPTOP/shun constant-level instrument); an electric constant temperature water bath (Tianjin Tiantai instruments ltd); YP5002 electronic balance (positron instruments ltd, city, changzhou); punch (diameter 8mm)

2.2 test methods

50 male ICR mice (18-20g) are adapted to the environment for 3 days, and are randomly divided into low, medium and high dose groups (2.6, 5.2 and 10.4g/kg) of the honeysuckle detoxification mixture, a model group and 10 aspirin groups (a positive drug group, 0.2g/kg) in each group according to the body mass. Gavage was administered 1 time per day for 7 consecutive days, and the blank control group was given equal volume of distilled water. Before the formal test, fasting is not forbidden for 12h, after the last administration is carried out for 0.5h, 50 mu L of dimethylbenzene is uniformly coated on the inner part and the outer part of the right auricle of a mouse, after 0.5h, the cervical vertebra of the mouse is dislocated and killed, 2 ears are cut along the baseline of the auricle, a double-ear piece is respectively punched at the same part by a puncher with the diameter of 8mm, the weight difference of the two ear pieces is used as swelling degree, and the swelling degree inhibition rate is calculated according to a formula.

2.3 test results

The results are detailed in table 2.

TABLE 2 Effect of flos Lonicerae detoxification mixture on ear swelling of mice caused by paraxylene

Note: compared with model group^#P＜0.05，^##P＜0.01

From the analysis in Table 2, the ear swelling degree of the mice in the aspirin, the middle-dose and high-dose combination of honeysuckle detoxification agent was significantly reduced (P < 0.05 or P < 0.01) compared with the model group.

(III) the antidotal mixture of the honeysuckle has the function of relieving cough: test for cough induced by strong ammonia water

3.1 materials and instruments

3.1.1 test animals

3.1.2 drugs and reagents

The content of the mixture is 0.5g per 1.0 ml. Storage conditions are as follows: refrigerating at 4 deg.C, storing under sealed condition, and re-heating to 37 deg.C for use in water bath during experiment; dextromethorphan hydrobromide tablets (Shijiazhuang, Ling pharmaceutical Co., Ltd., A1910011); ammonia (Hengxing chemical reagent manufacturing Co., Ltd., Tianjin, lot number: 20191215)

3.1.3 instruments and devices

FA2004 analytical balance (SOPTOP/shun constant-level instrument); an electric constant temperature water bath (Tianjin Tiantai instruments ltd); YP5002 electronic balance (positron instruments ltd, city, changzhou);

3.2 test methods

50 ICR mice (18-20g, half of each male and female) are adapted to the environment for 3 days, the grouping and administration mode of experimental animals is the same as 2.2, and only the mice with the positive medicine group are changed into gavage administration of dextromethorphan hydrobromide (18mg/kg) and continuous administration for 7 days. Fasting is not forbidden for 12h before the last administration, ammonia water is used for cough induction after the last administration is carried out for 0.5h, the mouse is placed in a 500ml glass bell cover which is placed upside down on a table, 0.5ml of concentrated ammonia water (25% -28% ammonium hydroxide) is blown on a cotton ball (100 +/-5 mg) by a liquid transfer gun, the mouse is immediately taken out after being placed in the bell cover for cough 30s, cough reflex (abdominal muscle contraction, mouth expansion and sometimes cough sound) of the mouse is observed, time from the time when the mouse is placed to the time when the mouse generates the cough reflex, namely cough latency and cough frequency of the mouse within 2min are recorded, and the cough inhibition rate is calculated according to a formula.

3.3 test results

The results are detailed in table 3.

Table 3 influence of antidotal mixture of flos coptidis on cough-inducing experiment with strong ammonia water in mice

Note: compared with model group^#P＜0.05，^##P＜0.01

The analysis of the table 3 shows that compared with the model group, the dextromethorphan hydrobromide and the high-dose flos coptidis detoxification mixture have obvious prolongation effect (P is less than 0.05 or less than 0.01) on the cough latent period of the mice with strong ammonia water induced cough, and the cough latent period of the mice in the low-dose and medium-dose flos coptidis detoxification mixture group also has a prolongation trend; compared with the model group, the cough frequency of mice in each dose group of dextromethorphan hydrobromide and the honeysuckle detoxification mixture is obviously reduced within 2min (P is less than 0.05 or P is less than 0.01), and each dose group presents better dose-effect relationship.

(IV) the antidotal mixture of the honeysuckle and the forsythia has the effect of eliminating phlegm: mouse tracheal phenol red excretion test

4.1 materials and instruments

4.1.1 test animals

4.1.2 drugs and reagents

The content of the mixture is 0.5g per 1.0 ml. Storage conditions are as follows: refrigerating at 4 deg.C, storing under sealed condition, and re-heating to 37 deg.C for use in water bath during experiment; ammonium chloride (Tianjin, Nature chemical Co., Ltd., Lot 20200122); phenol red (Tianjin new fine chemical development center, lot number: 20191211);

4.1.3 instruments and devices

FA2004 analytical balance (SOPTOP/shun constant-level instrument); an electric constant temperature water bath (Tianjin Tiantai instruments ltd); YP5002 electronic balance (positron instruments ltd, city, changzhou); TGL-16E desk top high speed refrigerated centrifuge (YINGTAI); an electric constant temperature water bath (Tianjin Tiantai instruments ltd); an enzyme-labeling instrument (BIO TEK company, model: Synergy MX);

4.2 test methods

50 ICR mice (18-20g, half of each male and female) are adapted to the environment for 3 days, the grouping and administration mode of experimental animals is the same as 2.2, and only the mice with the positive medicine group are changed into gavage administration of ammonium chloride (1g/kg) and continuous administration for 7 days. Fasting is not forbidden for 16h before last administration, after 0.5h of last administration, intraperitoneal injection is carried out for 0.2ml/10g of 2.5% phenol red physiological saline solution, after 0.5h of injection, cervical dislocation is carried out to kill animals, the animals are fixed in an upward position, the neck is straightened, the air outlet pipe is dissected and separated, a section of air pipe with the same length from the lower part of the thyroid cartilage to the branch part of the air pipe is cut off, the air pipe is placed into a test tube containing 2ml of 5% sodium bicarbonate, ultrasonic treatment is carried out for 30min, centrifugation is carried out for 5min at 2000 r/min, supernatant is taken, and an OD value is measured at 546nm by using an enzyme-labeling instrument.

4.3 test results

The results are detailed in table 4.

TABLE 4 Effect of antidotal mixture of Lonicera Japonica flos on mouse airway phenol Red excretion

Note: compared with model group^#P＜0.05，^##P＜0.01

The analysis in Table 4 shows that the mouse trachelol red excretion of the ammonium chloride group and the mouse trachelol red excretion of the honeysuckle detoxification mixture medium-dose and high-dose groups are obviously increased (P is less than 0.05 or P is less than 0.01) and the mouse trachelol red excretion of the honeysuckle detoxification mixture low-dose groups is also increased, but has no statistical significance (P is more than 0.05)

The toxicological demonstration of the honeysuckle detoxification mixture prepared by the invention is as follows.

Toxicity test of single administration of flos Coptidis detoxification mixture

1.1 materials and instruments

1.1.1 test animals

SPF KM mice 40, half male and female, body mass (20 + -2) g; purchased from Liaoning Experimental animals technology GmbH, quality certification number: SCXK (jew) 2020-: SYXK 2018-. SPF-level rat maintenance feed (batch No. 20111611) was purchased from Liaoning Biotechnology Ltd; poplar dunnage (performance standard QB2000) was purchased from seelboro biotechnology limited. The introduced water is sterilized tap water. An automatic timing light control system is adopted in the animal raising room to control the illumination period (12h light and shade alternation, illumination is started at 7:00 am every day), the raising temperature (24 +/-2) DEG C and the relative humidity of 50-60 percent. The experimental animals were acclimatized for 7 days in the above environment during which the animals had free access to food and water. The treatment of animals in the experimental process conforms to the guidance comments on the animals being studied in good care issued by the national department of science and technology in 2006.

1.1.2 drugs and reagents

The content of the mixture is 0.5g per 1.0 ml. Storage conditions are as follows: refrigerating at 4 deg.C, storing under sealed condition, and re-heating to 37 deg.C for use in water bath during experiment;

1.1.3 instruments and devices

FA2004 analytical balance (SOPTOP/shun constant-level instrument); an electric constant temperature water bath (Tianjin Tiantai instruments ltd); YP5002 electronic balance (positron instruments ltd, city, changzhou); TGL-16E desk top high speed refrigerated centrifuge (YINGTAI); an electric constant temperature water bath (Tianjin Tiantai instruments ltd);

1.2 test methods

1.2.1 preliminary experiments

20 KM mice, male and female. The groups were randomly divided into a blank control group and a honeysuckle detoxification mixture group, 10 of which were included in each group. Before the experiment, the fasting is not forbidden for 12 hours, the maximum concentration (3.2g/ml, the degree of a no-blockage No. 12 gavage needle) of the honeysuckle detoxification mixture for the gavage and the maximum volume (40ml/kg) of the gavage can be used for carrying out the gavage administration once, the mice in the blank control group are subjected to the gavage administration with the same volume of purified water, the death condition of the animals is observed after the administration, and the continuous observation is carried out for 14 days.

20 KM mice, male and female. The groups were randomly divided into a blank control group and a honeysuckle detoxification mixture group, 10 of which were included in each group. Before the experiment, the fasting is not forbidden for 12 hours, the maximum concentration (3.2g/ml, and the degree is that a No. 12 gavage needle is used) of the honeysuckle detoxification mixture for the gavage and the maximum volume (40ml/kg) of the mixture which can be borne by the gavage are used for carrying out the gavage and the administration twice, the mice in a blank control group are subjected to the gavage and the administration of purified water with the same volume, the death condition of the animals is observed after the administration, and the continuous observation is carried out for 14 days.

After 14 days observation, the mice were all alive. The oral gavage dose, limited by the dose concentration and dose volume, failed to measure half the Lethal Dose (LD) of the honeysuckle detoxification mixture₅₀) Therefore, the maximum dose test of the honeysuckle detoxification mixture is carried out.

1.2.1 official test

40 KM mice, each half male and female, were randomly divided into a blank control group and a honeysuckle detoxification mixture group, and 20 mice were used in each group. Before the experiment, the mice are fasted and not forbidden to be watered for 12 hours, and the highest concentration (3.2g/ml) and the maximum administration volume (40ml/kg) of the medicine are intragastrically administered for 2 times within 24 hours, the two administration intervals are 6 hours, and the mice in a blank control group are intragastrically administered with the purified water with the same volume. After administration, the animals were observed mainly for death and toxic reactions (including hair color, autonomic activity, abnormal movement, response to stimulation, respiration and its frequency, oral and nasal secretions, stool characteristics, perineal secretions, external genitalia, eyeball, etc.), and then general observations were continued for 14 days. The animal body is weighed 7 days and 14 days after administration, gross dissection is carried out after the observation period is finished, and the histopathological examination is carried out on the organ with pathological changes if the major organs and tissues such as liver, heart, spleen, lung, kidney, adrenal gland, thymus, stomach and intestine, testis (male), epididymis (male), uterus (female), ovary (female) and the like are mainly observed, if the organs have macroscopic changes in volume, color, texture and the like.

1.3 general case observations

General conditions of mice: after the honeysuckle detoxification mixture with the maximum concentration and the maximum administration volume is infused for 1 hour, the mice in the administration group have reduced autonomous activity, and loose stool and unformed stool appear after 3 hours of infusion, but the color of the stool is normal, the mice in the administration group recover to eat after 3 hours of second-time infusion administration, the ingestion behaviors of the animals in the administration group are reduced, the ingestion behaviors and activities gradually recover to normal after 6 hours of administration, and the stools recover to normal the next day. In the observation period, the central nervous system, the somatic movement, the autonomic nervous system, the respiratory system, the skin, the hair and the like of the mice in the administration group are not abnormal, and the mice are not dead. The changes in body mass of mice are detailed in Table 5. Before and 7 days and 14 days after the administration, the body mass of two groups of mice has no significant difference.

TABLE 5 mice pre-dose and 7 days, 14 days body mass changes

1.4 systematic necropsy

And C, a dissection examination result: the brain, liver, heart, spleen, lung, kidney, adrenal gland, thymus, testis, epididymis, uterus and ovary of the mice in the blank group and the administration group have no macroscopic pathological changes of size, color, texture and the like.

(II) repeated administration toxicity test of flos Coptidis detoxification mixture

2.1 materials and instruments

2.1.1 test animals

SPF SD rats 80 with half male and female, body mass (140-: SCXK (jew) 2020-: SYXK 2018-.

SPF-level rat maintenance feed (batch No. 20111611) was purchased from Liaoning Biotechnology Ltd; poplar dunnage (performance standard QB2000) was purchased from seelboro biotechnology limited. The drinking water is sterilized tap water. The animal raising room adopts an automatic timing light control system to control the illumination period (12h light and shade alternation), the raising temperature (24 +/-2) DEG C and the relative humidity of 50-60 percent. The experimental animals were acclimatized for 7 days in the above environment during which the animals had free access to food and water. The treatment of animals in the experimental process conforms to the guidance comments on the animals being studied in good care issued by the national department of science and technology in 2006.

2.1.2 drugs and reagents

2.1.3 instruments and apparatus

FA2004 analytical balance (SOPTOP/shun constant-level instrument); an electric constant temperature water bath (Tianjin Tiantai instruments ltd); YP5002 electronic balance (positron instruments ltd, city, changzhou); XN-1000 full-automatic hemocyte analyzer (Japanese sysmex); AU5811 fully automated biochemical analyzer (BECKMAN COULTER); pure water system (ELGA); a biological tissue roast machine (Sudoku medical electronic technology Co., Ltd., Miniaoling city); HM340E paraffin microtome (semer feishell instruments ltd); HM525 NX U cryomicrotome (semer feishel instruments ltd); CX41RF optical microscope (OLYMPUS); TGL16E desk type high speed refrigerated centrifuge (Changsha Yingtai apparatus Co., Ltd.)

2.2 test methods

2.2.1 grouping and administration

Rats are randomly divided into low, medium and high dose groups of the honeysuckle detoxification mixture (the administration doses are respectively 0.8g/kg, 1.6g/kg and 3.2g/kg in crude drug count, which are respectively equal to 14 times, 28 times and 56 times of clinical medication, and the dose design is detailed in table 6) and blank control groups (purified water is given), wherein each group comprises 20 rats, each male and female half, each group of rats is subjected to intragastric administration once according to the corresponding dose, the intragastric volume is 10ml/kg, and the administration is continuously carried out for 30 days. After the administration and 14 days (recovery period), 10 animals (female and male half) are randomly selected from each group, fasting is not forbidden for 12 hours, 10% chloral hydrate is injected into the abdominal cavity for anesthesia, blood is collected through abdominal aorta, and hematology, hematobiochemistry and histopathology index detection is carried out.

TABLE 6 LONG-TERM TOXICITY TEST DOSE DESIGN OF LIAN HUA DETOXICATION AGENT

2.3 general case observations

During the administration period, the animals were observed for their appearance of hair, secretions, behavioral activities, respiration, stool characteristics, toxic reactions, death, and the like.

During the period of adapting to the environment, the behavior, activity, ingestion, drinking, stool character and other conditions of the animals in the blank group and the low, medium and high dose groups of the honeysuckle detoxification mixture are normal, and the tested animals have no abnormal performance, which indicates that the selected tested animals meet the requirements of long-term toxicity tests. After oral gavage, the animals in the high dose group had loose stools and abnormal stools on days 2 and 3, and recovered to normal on day 4. The other groups of animals tested did not find any abnormal condition. The recovery animals have no abnormal condition.

2.4 measurement of food intake

The daily intake of rats was measured and the results are shown in tables 7 and 8

TABLE 7 Effect on average food intake in rats during dosing period

TABLE 8 Effect on average food intake in convalescent rats

As analyzed in tables 7 and 8, the food intake of male and female rats in each administration group was not significantly different from that in the blank group by P > 0.05 at the end of the administration period. At the end of the withdrawal recovery period, the food intake of male and female rats in each administration group is not significantly different from that in the blank group, and P is more than 0.05.

2.5 measurement of body Mass

The body weight was weighed every week, and the administration amount was adjusted according to the measured body constitution, and the change in body constitution is shown in Table 9, Table 10, Table 9, the influence on the body mass of rats in the administration period

Note: p < 0.05 in comparison with blank

TABLE 10 Effect on rat body Mass during recovery

As analyzed in tables 9 and 10, the body mass of the male rats in the high dose group was reduced (P < 0.05) compared with that of the male rats in the blank control group at the third week of administration, and the body mass of the male and female rats in the remaining time points of administration was not significantly different (P > 0.05) compared with that of the blank group. During the drug withdrawal recovery period, the body mass of male and female rats in each administration group is not significantly different from that in the blank group (P > 0.05).

2.6 measurement of organ coefficients

The rats are subjected to systematic autopsy at the end of the administration period and at the end of the recovery period, the main organs are weighed, and the organ coefficients are calculated according to a formula. The test results are shown in tables 11 to 14

TABLE 11 Effect on organ coefficients of Male rats 1

TABLE 12 Effect on organ coefficients of Male rats 2

TABLE 13 Effect on organ coefficients of female rats 1

TABLE 14 Effect on organ coefficients of female rats 2

From the analysis in tables 11 to 14, the organ coefficients of the male rats of the administration groups at the end of the administration period and the end of the withdrawal recovery period were not significantly different from those of the blank group by P > 0.05. Compared with the blank group, the organ coefficient of the female rats of each administration group at the end of the administration period and the end of the withdrawal recovery period has no significant difference P larger than 0.05.

2.7 hematological examination

White Blood Cells (WBC), Neutrophils (NEU), lymphocyte counts (LYM), Monocytes (MON), Eosinophils (EOS), Red Blood Cells (RBC), Hemoglobin (HGB), mean volume of red blood cells (MCV), mean hemoglobin amount (MCH), mean hemoglobin concentration (MCHC), width of red blood cell distribution CV (RDW-CV), Platelets (PLT), width of Platelet Distribution (PDW), Mean Platelet Volume (MPV). The test results are detailed in tables 15-22.

TABLE 15 Effect on hematological indices in Male rats at the end of dosing period 1

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

TABLE 16 Effect on hematological indices in Male rats at the end of dosing period 2

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

TABLE 17 Effect on hematological indices in female rats at the end of dosing period 1

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

TABLE 18 Effect on hematological indices in female rats at the end of dosing period 2

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

TABLE 19 Effect on hematological indices in Male rats at the end of withdrawal recovery period 1

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

TABLE 20 Effect on hematological indices in Male rats at the end of withdrawal recovery period 2

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

TABLE 21 Effect on hematological indices in female rats at the end of withdrawal recovery period 1

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

TABLE 22 Effect on hematological indices in female rats at the end of withdrawal recovery period 2

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

At the end of the administration period, compared with the blank group of male rats, the mononuclear cell number of the male rats in the low-dose group and the platelet number and the mean hemoglobin amount of the male rats in the medium-dose group are obviously increased (P < 0.05 or P < 0.01), the mean volume of the red blood cells of the male rats in the low-dose group and the middle-dose group is obviously increased (P < 0.05), and the hemoglobin level of the red blood cells of the male rats in the high-dose group and the red blood cells of the male rats in the medium-dose group are obviously reduced (P < 0.05 or P < 0.01). The other groups showed no difference in hematological index compared to the blank control group.

At the end of the withdrawal recovery period, the mean hemoglobin concentration of the medium-dose group female rats was significantly reduced (P < 0.05) and the mean hemoglobin concentration of the high-dose group female rats was significantly increased (P < 0.05) compared to the blank-control group female rats. The other groups showed no difference in hematological index compared to the blank control group.

2.8 serum chemistry examination

Aspartate Aminotransferase (AST), alanine Aminotransferase (ALT), alkaline phosphatase (ALP), Total Protein (TP), Albumin (ALB), Globulin (GLB), Urea (Urea), Creatinine (CREA), Glucose (GLU), Triglyceride (TG), Cholesterol (CHO), potassium (K), sodium (Na), Chlorine (CL). The test results are detailed in tables 23 to 30.

TABLE 23 Effect on serum Biochemical indicators of Male rats at the end of dosing period 1

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

TABLE 24 Effect on serum Biochemical indicators in Male rats at the end of dosing period 2

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

TABLE 25 Effect on serum Biochemical indicators of female rats at the end of dosing period 1

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

TABLE 26 Effect on serum Biochemical indicators of female rats at the end of dosing period 2

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

TABLE 27 Effect on Biochemical indicators of serum from Male rats at the end of drug withdrawal period 1

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

TABLE 28 Effect on serum Biochemical indicators of Male rats at the end of drug withdrawal period 2

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

TABLE 29 Effect on serum Biochemical indicators of female rats at the end of drug withdrawal period 1

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

TABLE 30 Effect on serum Biochemical indicators of female rats at the end of drug withdrawal period 2

Note: p < 0.05 compared to blank; comparison with blank group, P < 0.01

At the end of the administration period, compared with the blank group of male rats, the serum creatinine and potassium ion levels of the male rats in the low-dose group are obviously increased (P < 0.05 or P < 0.01), the serum sodium and urea levels of the male rats in the low-dose group are obviously reduced (P < 0.05) compared with the serum urea levels of the male rats in the high-dose group, and the serum chloride ion levels of the male rats in the medium-dose group and the high-dose group are extremely obviously increased (P < 0.01). At the end of the administration period, compared with a blank group of female rats, the serum alkaline phosphatase level of the female rats in the medium-dose group is obviously increased (P < 0.05), the serum glucose level of the female rats in the low-dose group, the medium-dose group and the high-dose group is obviously increased (P < 0.01), the triglyceride level of the female rats in the low-dose group is obviously increased (P < 0.01), and the serum potassium ion level of the female rats in the medium-dose group and the high-dose group is obviously increased (P < 0.05). The biochemical indexes of the sera of the other groups are not different from those of the blank control group.

At the end of the drug withdrawal period, compared with male rats in a blank control group, the serum alkaline phosphatase and urea of male rats in a low dose group are remarkably increased (P < 0.05 and P < 0.01), the serum creatinine level of male rats in a low dose group and a medium dose group is remarkably increased (P < 0.01 and P < 0.05), the serum glucose level of male rats in a low dose group and a high dose group is remarkably increased (P < 0.05), the serum triglyceride and sodium ion level of male rats in a low dose group is remarkably reduced (P < 0.05), and the serum potassium ion and chloride ion level of male rats in a medium dose group is remarkably reduced (P < 0.05). Compared with the blank control group female rats, the serum sodium ion level of the female rats in the low-dose group is obviously increased (P < 0.05). The serum triglyceride level of female rats in the medium dose group is obviously increased (P < 0.05). The biochemical indexes of the sera of the other groups are not different from those of the blank control group.

2.9 systematic necropsy and histopathology

Performing systematic dissection, observing whether brain, thymus gland, heart, lung, liver, spleen, kidney, adrenal gland, testis, epididymis, uterus, and ovary have obvious volume, color, and texture changes, fixing with 4% paraformaldehyde solution, and performing histopathological examination

Brain: blank group: after 30 days of administration and at the end of the recovery period, the animal brain cells have normal morphology, and the neuron cells have no degeneration, necrosis, hemorrhage, infarction and space occupying lesion. High dose group: after 30 days of administration and at the end of the recovery period, no lesion is observed in the brain tissue.

Liver: the hepatic lobules of the blank group and the high-dose group are clear and complete in structure, the liver chordae morphology and distribution are normal, the liver cell nucleus is located in the center, cytoplasm is uniformly red-stained, and no vacuole or obvious inflammatory reaction exists in the cell.

Heart: blank group: the myocardial cell cytoplasm is red stained, the texture is clear, and edema, steatosis or necrosis are not seen. The intercalated disc between the cardiac muscle cells is invisible; the interstitial substance of the cardiac muscle has no inflammatory cells, fat infiltration and fiber hyperplasia, the administration lasts for 30 days and at the end of the recovery period, and the cardiac muscle tissue has no pathological changes. The high dose group is administrated for 30 days and the recovery period is over, and no pathological changes are found in the myocardial tissues.

Spleen: the tissue morphological structure of the blank group and the high-dose group is normal, the boundary of the white marrow region is clear, the lymphocyte arrangement of the white marrow region is compact, the structure of the lymph follicle is clear and visible, and the red marrow region is rich in the blood sinuses filled with red blood cells.

Lung: from the appearance, both the blank and high dose groups had a pale lung with a soft texture and no significant shrinkage. HE staining shows that the alveoli of the blank group and the high-dose group are clear, complete and uniform in size, rich in alveoli, smooth and thin in alveolar wall, complete in structure, free of inflammatory cell infiltration in pulmonary interstitium, and free of congestion and hematoma in the alveolar.

Kidney: blank group: the renal tubular epithelial cells have clear structures and no obvious degeneration and necrosis. High dose group: after 30 days of administration and at the end of the recovery period, no lesion was observed in the kidney tissue.

Adrenal gland: blank group: the structure of adrenal cortex and medulla is clear, the adrenal gland tissue has no pathological changes after 30 days of administration and at the end of recovery period. High dose group: the adrenal gland tissue has no pathological changes after 30 days of administration and at the end of the recovery period.

Thymus: the thymus tissue structures of the blank group and the high-dose group are clear, the cortex medulla demarcation is obvious, the arrangement of the lymphocyte in the cortex area is regular and compact, and the thymocytes can be seen in the medulla area.

Testis: blank group: after 30 days of administration and the recovery period, the seminiferous tubular structure is clear, the spermatogenic cells at all levels are normal in shape, the spermatogenic cells are clear and visible, and the supporting cells are not abnormal; the structures under testis and interstitial mirror are approximately normal. High dose group: after 30 days of administration and the recovery period, no lesion was observed in the testis tissue.

Epididymis: blank group: after 30 days of administration and the recovery period, the epithelial cells of the output tubule and the epididymis duct have clear shapes, no inflammatory lesion is seen, and no inflammatory infiltration is seen in the interstitium. High dose group: after 30 days of administration and the recovery period, no lesion is found in the epididymis tissue.

Uterus: blank group: after 30 days of administration and the recovery period, no lesion was observed in the uterine tissue. High dose group: after 30 days of administration and the recovery period, no lesion was observed in the uterine tissue.

Ovary: blank group: after 30 days of administration and the recovery period, the ovarian skin and medulla structures are clear, all levels of follicles are clear and visible, and the shape and the number of the follicles in each period are normal; the primary oocyte belongs to a normal cell morphological structure; part of the ovary has no follicular chocolate cyst or blood clot. High dose group: after 30 days of administration and the recovery period, no lesion was observed in ovarian tissue.

Trachea: in appearance, the blank group and the high-dose group both have clear and transparent trachea appearance and good touch toughness. HE staining showed that both the blank and high dose groups had intact tracheal mucosal epithelium, no exfoliated cells in the lumen, and no inflammatory cell infiltration in the interstitium.

Duodenum: blank group: after 30 days of administration and the recovery period, the duodenum mucosa epithelium has no degeneration, necrosis, ulcer, hemorrhage, and inflammation. High dose group: after 30 days of administration and the recovery period, no lesions were observed in the duodenal tissues. (see the drawings for details, the pictures are all taken under a 400-fold mirror), and the results are detailed in fig. 1-4.

The above embodiments are merely preferred embodiments of the present disclosure, which are not intended to limit the present disclosure, and any modifications, equivalents, improvements and the like, which are within the spirit and principle of the present disclosure, should be included in the scope of the present disclosure.

Claims

1. A preparation process of a honeysuckle detoxification mixture capable of improving immunity is characterized by comprising the following steps: the Chinese medicinal preparation is prepared from the following raw material medicaments in percentage by weight (in terms of prepared 1000ml of medicaments): 18.75 percent of honeysuckle, 18.75 percent of forsythia, 18.75 percent of atractylodes, 12.5 percent of platycodon root, 12.5 percent of mulberry leaf, 12.5 percent of eupatorium and 6.25 percent of liquorice;

soaking the above seven materials in water, decocting, standing, filtering, concentrating the filtrate under reduced pressure to 1000ml, and packaging.

2. The preparation process of the honeysuckle detoxification mixture capable of improving immunity as claimed in claim 1, wherein the preparation process comprises the following steps: the specific weight of the raw material medicaments is as follows: 37.5g of honeysuckle, 37.5g of fructus forsythiae, 37.5g of bighead atractylodes rhizome, 25g of platycodon grandiflorum, 25g of mulberry leaf, 25g of eupatorium and 12.5g of liquorice.

3. The preparation process of the honeysuckle detoxification mixture capable of improving immunity as claimed in claim 1, wherein the preparation process comprises the following steps: the soaking time in water is 1 hour, the decocting time is 40 minutes, and the net standing time is 24 hours.

4. The preparation process of the honeysuckle detoxification mixture capable of improving immunity as claimed in claim 1, wherein the preparation process comprises the following steps: and adding sodium benzoate into the concentrated solution, and uniformly mixing.