Application of pine pollen in preparing a preparation for preventing or treating cadmium exposure and method for promoting cadmium excretion by an organism

Abstract

The present invention provides an application of pine pollen in the preparation of a preparation for preventing or treating cadmium exposure and a method for promoting cadmium excretion from the body, which relates to the technical field of traditional Chinese medicine. In a cadmium-exposed animal model intervened with pine pollen, for the organs of the kidney, testis and liver that are prone to cadmium accumulation upon cadmium exposure, the cadmium content therein significantly decreases, and to a certain extent, it can protect the morphological integrity of the kidney, and significantly improve the damage to testicular tissue caused by cadmium exposure and alleviate tissue morphological abnormalities. Pine pollen can also significantly increase the urinary cadmium and fecal cadmium excretion amounts in rats with chronic cadmium exposure. Applying pine pollen to the preparation of a preparation for preventing or treating cadmium exposure alleviates the current increasing problem of cadmium exposure risk.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and particularly relates to the application of pine pollen in the preparation of a preparation for preventing or treating cadmium exposure and a method for promoting cadmium excretion from the body. Background Art

[0002] Cadmium (Cd) is a colored heavy metal element that can be toxic to multiple organs of humans and animals and is one of the common environmental and industrial pollutants. Since cadmium in the environment cannot be biodegraded, the content of cadmium in water bodies and soil increases year by year, and then accumulates in some plants or animals. In addition to accumulating in the human body through the food chain, smoking is also an important route of cadmium exposure.

[0003] Cadmium is slowly metabolized in the body and has an extremely long half-life, about 10 - 36 years, which is the main cause of chronic cadmium poisoning. Long-term accumulation of trace amounts of cadmium intake will seriously affect the health of the body. The most serious health effect of cadmium is its impact on bones. Cadmium accumulates in bones, which will directly lead to weakened bone brittleness and cause osteomalacia and osteoporosis. Secondly, the health effect of cadmium poisoning is the damage to reproductive organs. Long-term cadmium exposure can lead to abnormal development and function of testicular cells, induce vacuole formation in the spermatogenic epithelium, abnormal changes in the structure of testicular Sertoli cells, damage the blood-testis barrier, increase the permeability of seminiferous tubules, reduce testosterone secretion, interfere with the development and function of germ cells. Eventually, cadmium will lead to abnormal sperm morphology and reduced sperm motility. In addition to the damage to bones and the reproductive system, long-term accumulation of cadmium in the body will also cause damage to the kidneys, liver, lungs, and immune system. Therefore, how to reduce the harm caused by cadmium to the body is a problem to be solved at present.

[0004] Therefore, how to reduce the harm caused by cadmium to the body is a problem to be solved at present.

[0005] In view of this, the present invention is specifically proposed. Summary of the Invention

[0006] The first object of the present invention is to provide an application of pine pollen in the preparation of a preparation for preventing or treating cadmium exposure to alleviate the current increasing risk of cadmium exposure.

[0007] The second object of the present invention is to provide a method for promoting cadmium excretion from the body for non-diagnostic and therapeutic purposes.

[0008] To solve the above technical problems, the present invention specifically adopts the following technical solutions:

[0009] According to one aspect of the present invention, the present invention provides an application of pine pollen in the preparation of a preparation for preventing or treating cadmium exposure.

[0010] Preferably, the cadmium exposure includes chronic cadmium exposure.

[0011] Preferably, the subject to which the preparation is administered includes mammals;

[0012] Preferably, the mammals include mice, rats, guinea pigs, hamsters, rabbits, ferrets, cats, dogs, goats, sheep, cows, pigs, horses, monkeys or humans;

[0013] Preferably, the mammals include humans or rats;

[0014] Preferably, the mammals include cadmium-exposed animal models.

[0015] Preferably, the cadmium-exposed animal model includes a chronic cadmium-exposed rat model, and the chronic cadmium-exposed rat model is a rat with a CdCl2 injection dose of 0.1-0.4 mg / kg body weight;

[0016] Preferably, the subject to which the preparation for preventing or treating cadmium exposure is administered is the chronic cadmium-exposed rat model. Based on the amount of pine pollen in the preparation, the daily intake dose of the preparation is 0.2-2 g / kg body weight.

[0017] Preferably, the subject to which the preparation for preventing or treating cadmium exposure is administered is the chronic cadmium-exposed rat model. Based on the amount of pine pollen in the preparation, the daily intake dose of the preparation is 0.2-0.6 g / kg body weight.

[0018] Preferably, the subject to which the preparation for preventing or treating cadmium exposure is administered is humans. Based on the amount of pine pollen in the preparation, the daily intake dose of the preparation is 1.2-12 g / kg body weight.

[0019] Preferably, the preparation for preventing or treating cadmium exposure includes a single-agent preparation or a compound preparation;

[0020] Preferably, the preparation for preventing or treating cadmium exposure includes capsules, tablets, granules, pills, dripping pills, lyophilized powder, syrups or oral liquids;

[0021] Preferably, the preparation for preventing or treating cadmium exposure includes a sustained-release preparation or a controlled-release preparation.

[0022] According to another aspect of the present invention, the present invention also provides a method for promoting cadmium excretion from the body for non-diagnostic and therapeutic purposes. The method includes administering pine pollen or the above-mentioned preparation for preventing or treating cadmium exposure to the body.

[0023] Preferably, the method includes constructing a cadmium-exposed animal model and feeding pine pollen to the cadmium-exposed animal model;

[0024] Preferably, the method includes constructing a chronic cadmium-exposed rat model.

[0025] Preferably, the establishment of the chronic cadmium-exposed rat model includes injecting CdCl2 at a dose of 0.1-0.4 mg / kg body weight into the rats.

[0026] Preferably, the daily intake dose of pine pollen is 0.2-2 g / kg body weight.

[0027] Preferably, the daily intake dose of pine pollen is 0.2-0.6 g / kg body weight.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] Through experiments, the present invention found that in the cadmium-exposed animal model intervened by pine pollen, for the organs of the kidneys, testes and liver that are prone to cadmium accumulation, the cadmium content therein decreased significantly, and to a certain extent, it can protect the morphological integrity of the kidneys, and significantly improve the damage of testicular tissue caused by cadmium exposure, and reduce tissue morphological abnormalities. Pine pollen can also significantly increase the urinary cadmium and fecal cadmium excretion in chronic cadmium-exposed rats.

[0030] Using pine pollen to prepare a preparation for preventing or treating cadmium exposure can enable the preparation to have a cadmium-excreting effect, and can protect the health of the body's liver and kidney organs and reproductive system by promoting the cadmium-excreting effect, and enhance the activity of antioxidant enzymes. Pine pollen is a substance homologous to medicine and food, with high safety, and is a good choice for effectively removing heavy metal cadmium. Using pine pollen for promoting the body to excrete cadmium for non-diagnostic and therapeutic purposes can effectively promote the excretion of cadmium in the body by the subjects, and can be widely applied to the establishment of animal models, drug research and development, and the research on the physiological and biochemical functions related to the pathogenesis mechanism of cadmium, which is helpful for the in-depth research on the pathogenesis mechanism of cadmium in this field. Description of the Drawings

[0031] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required to be used in the description of the specific embodiments or the prior art. Obviously, the following drawings are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0032] Figure 1 Effects of pine pollen on the body weight of chronic cadmium-exposed rats, where CON is the normal group; CdE is the cadmium-exposed group; PP-H is the high-dose pine pollen group; PP-M is the medium-dose pine pollen group; PP-L is the low-dose pine pollen group;

[0033] Figure 2AEffect of pine pollen on the content of glutathione peroxidase in rats with chronic cadmium exposure. The values are expressed as mean ± standard deviation, n = 8 - 12; where CON is the normal group; CdE is the cadmium exposure group; PP-H is the high-dose pine pollen group; PP-M is the medium-dose pine pollen group; PP-L is the low-dose pine pollen group;

[0034] Figure 2B Effect of pine pollen on the lipid peroxidation product MDA in rats with chronic cadmium exposure. The values are expressed as mean ± standard deviation, n = 8 - 12; *P < 0.05, compared with the model group at the same time period; where CON is the normal group; CdE is the cadmium exposure group; PP-H is the high-dose pine pollen group; PP-M is the medium-dose pine pollen group; PP-L is the low-dose pine pollen group;

[0035] Figure 3 Effect of pine pollen on the fecal cadmium excretion in rats with chronic cadmium exposure. The values are expressed as mean ± standard deviation, n = 8 - 12, #P < 0.05, ##P < 0.01, compared with the normal group at the same time period; *P < 0.05, **P < 0.01, compared with the model group at the same time period; where CON is the normal group; CdE is the cadmium exposure group; PP-H is the high-dose pine pollen group; PP-M is the medium-dose pine pollen group; PP-L is the low-dose pine pollen group;

[0036] Figure 4 Effect of pine pollen on the urinary cadmium excretion in rats with chronic cadmium exposure. The values are expressed as mean ± standard deviation, n = 8 - 12, #P < 0.05, ##P < 0.01, compared with the normal group at the same time period; *P < 0.05, **P < 0.01, compared with the model group at the same time period; where CON is the normal group; CdE is the cadmium exposure group; PP-H is the high-dose pine pollen group; PP-M is the medium-dose pine pollen group; PP-L is the low-dose pine pollen group;

[0037] Figures 5A to 5D Effect of pine pollen on the cadmium in the liver, kidney, testis and femur of rats with chronic cadmium exposure; Figure 5A Results of cadmium changes in the liver, Figure 5B Results of cadmium changes in the kidney, Figure 5C Results of cadmium changes in the testis, Figure 5D Results of cadmium changes in the femur; The values are expressed as mean ± standard deviation, n = 8 - 12, #P < 0.05, ##P < 0.01, compared with the normal group at the same time period; *P < 0.05, **P < 0.01, compared with the model group at the same time period; where CON is the normal group; CdE is the cadmium exposure group; PP-H is the high-dose pine pollen group; PP-M is the medium-dose pine pollen group; PP-L is the low-dose pine pollen group;

[0038] Figure 6 A and Figure 6B shows the effects of pine pollen on the morphology of the kidneys and testes of rats with chronic cadmium exposure (×200); among them Figure 6 A is the experimental result of HE staining of rat testes; Figure 6 B is the experimental result of HE staining of rat kidneys; n = 7 - 9; among them, CON is the normal group; CdE is the cadmium exposure group; PP-H is the high-dose pine pollen group; PP-M is the medium-dose pine pollen group; PP-L is the low-dose pine pollen group;

[0039] Figures 7A to 7D Shows the effects of pine pollen on zinc in the liver, kidneys, testes and femurs of rats with chronic cadmium exposure. Among them Figure 7A Are the results of zinc changes in the liver, Figure 7B Are the results of zinc changes in the kidneys, Figure 7C Are the results of zinc changes in the testes, Figure 7D Are the results of zinc changes in the femurs. The values are expressed as mean ± standard deviation, n = 8 - 12, #P < 0.05, ##P < 0.01, compared with the normal group in the same period; *P < 0.05, **P < 0.01, compared with the model group in the same period; among them, CON is the normal group; CdE is the cadmium exposure group; PP-H is the high-dose pine pollen group; PP-M is the medium-dose pine pollen group; PP-L is the low-dose pine pollen group;

[0040] Figures 8A to 8B Shows the effects of pine pollen on fecal zinc and urinary zinc in rats with chronic cadmium exposure; among them Figure 8A Are the results of fecal zinc changes, Figure 8B Are the results of urinary zinc changes. The values are expressed as mean ± standard deviation, n = 8 - 12, #P < 0.05, ##P < 0.01, compared with the normal group in the same period; *P < 0.05, **P < 0.01, compared with the model group in the same period; among them, CON is the normal group; CdE is the cadmium exposure group; PP-H is the high-dose pine pollen group; PP-M is the medium-dose pine pollen group; PP-L is the low-dose pine pollen group. Specific implementation manners

[0041] Next, the technical solutions of the present invention will be clearly and completely described in conjunction with the embodiments. Obviously, the described embodiments are some embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0042] Pine pollen refers to the dried pollen of Pinus massoniana, Pinus tabuliformis or plants of the same genus in the Pinaceae family. The medicinal and edible value of pine pollen has a long history in traditional Chinese medicine. It is one of the two kinds of pollen recorded in ancient Chinese medicine books. It is described in works such as "Shennong Ben Cao Jing", "Compendium of Materia Medica", and "New Compilation of Materia Medica", and is called: "Nourishing the heart and lungs, replenishing qi, dispelling wind and stopping bleeding". Pine pollen is rich in more than 200 kinds of nutritional and active ingredients such as polysaccharides, proteins, amino acids, unsaturated fatty acids, vitamins, minerals, and various bioactive ingredients such as flavonoids, choline, and phytosterols. It is rich in nutrition and easy to absorb, so it is known as the "miniature nutrient library". Pine pollen has significant medicinal and health care effects. A large number of studies have shown that pine pollen has various health care functions such as enhancing immunity, antioxidant, anti-aging, anti-tumor, hypoglycemic, lipid-lowering, preventing and treating cardiovascular and cerebrovascular diseases, enhancing physical endurance, regulating the gastrointestinal microecological environment, protecting internal organs, inhibiting the body's hormone levels, and anti-inflammatory and antipruritic.

[0043] So far, there has been no relevant verification study on the cadmium excretion function of pine pollen. Through relevant experimental studies, this invention verifies with multiple indicators that pine pollen has the function of cadmium excretion and can be used to prepare preparations for preventing or treating cadmium exposure.

[0044] The inventor first established a chronic cadmium exposure model in rats by injecting rats with low-dose cadmium chloride (CdCl2) for a long time. The CdCl2 injection dose was 0.1 - 0.4 mg / kg body weight to establish a chronic cadmium exposure model in rats.

[0045] Observing the effect of pine pollen on the body weight of rats with chronic cadmium exposure, the results showed that chronic cadmium exposure and pine pollen intervention had no obvious effect on the body weight of rats.

[0046] Observing the effect of pine pollen on antioxidant enzymes in rats with chronic cadmium exposure, the experimental results showed that pine pollen could significantly increase the activity of glutathione peroxidase (GSH-Px) and significantly reduce the production of peroxidation product MDA in cadmium-exposed rats.

[0047] Using atomic absorption spectrophotometry to observe the cadmium element content in the liver, kidney, testis and femur of rats with chronic cadmium exposure after pine pollen intervention. The experimental results showed that pine pollen had a very significant cadmium excretion effect, and the cadmium content in the organs such as the liver, kidney, testis and femur where cadmium is easily accumulated decreased significantly.

[0048] Using atomic absorption spectrophotometry to observe the cadmium element excretion in feces and urine of rats with chronic cadmium exposure after pine pollen intervention. The experimental results showed that pine pollen intervention could significantly increase the cadmium excretion in urine and feces of rats with chronic cadmium exposure, and the effect showed a positive dose-dependent effect. Urinary cadmium excretion is the main way of cadmium excretion, and the urinary cadmium excretion accounts for more than 90% of the total cadmium excretion.

[0049] The hematoxylin-eosin staining method (HE staining) was used to observe the effects of pine pollen on the morphology of the testis and kidney in rats with chronic cadmium exposure. The experimental results showed that chronic cadmium exposure caused certain damage to the kidneys of rats, and pine pollen intervention could protect the morphological integrity of the kidneys to a certain extent. Chronic cadmium exposure caused very serious structural abnormalities in the testis of rats, the absence of spermatocytes, disordered arrangement of spermatogonia, a significant decrease in the number, and a large amount of protein mucus was visible in the lumen. Pine pollen intervention could significantly improve the damage to testicular tissue caused by cadmium exposure and reduce tissue morphological abnormalities.

[0050] Flame atomic absorption spectrometry was used to observe the effects of pine pollen on zinc in rats with chronic cadmium exposure. The experimental results showed that the zinc content in the kidneys and testis of the rats in the chronic cadmium exposure model group was significantly lower than that in the control group (p<0.05). After pine pollen intervention, the zinc content in the organs increased very significantly compared with the model group and was similar to the control group level. Pine pollen intervention could inhibit the loss of zinc in rats caused by chronic cadmium exposure and did not promote zinc excretion.

[0051] The above experimental results indicate that pine pollen has a cadmium-excreting effect, can protect the health of the liver, kidney organs and reproductive system by promoting cadmium excretion, and enhance the activity of antioxidant enzymes. Pine pollen is a substance with both medicinal and edible properties, has high safety, and is a good choice for effectively removing heavy metal cadmium.

[0052] Based on the above experimental findings of the inventors, the present invention provides an application of pine pollen in the preparation of a preparation for preventing or treating cadmium exposure. The preparation for preventing or treating cadmium exposure according to the present invention can be used for the prevention, alleviation or treatment of cadmium exposure; or for assisting in the prevention, alleviation or treatment of cadmium exposure; or for promoting the excretion of cadmium from the body of subjects in scientific research. Preparations with the above functions all belong to the preparation for preventing or treating cadmium exposure. The preparation for preventing or treating cadmium exposure includes but is not limited to experimental reagents, traditional Chinese medicine compositions or further processed traditional Chinese medicine preparations, health products or foods. The specific indications of the preparation for preventing or treating cadmium exposure include but are not limited to reproductive diseases such as weakened bone brittleness, osteomalacia, osteoporosis, testicular dysfunction, etc. caused by long-term accumulation of trace cadmium intake, kidney, liver, lung or immune system damage, or tumors.

[0053] The preparation for preventing or treating cadmium exposure is preferably used for preventing, alleviating or treating chronic cadmium exposure. Chronic cadmium exposure refers to the phenomenon that cadmium enters the human body through occupational exposure, dietary intake, cigarette smoke and environmental skin contact in an environment containing cadmium, thereby causing chronic cadmium poisoning. The chronic cadmium exposure can be defined as is well known in the art and as recorded in various general and more specific textbooks, reference documents and standard documents. For example, according to the symptoms caused by generally preparing a chronic cadmium exposure animal model in the art, or in line with the symptoms of clinical chronic cadmium poisoning.

[0054] The administration subjects of the preparation for preventing or treating cadmium exposure include, but are not limited to, mammals, such as, but not limited to, mice, rats, guinea pigs, hamsters, rabbits, ferrets, cats, dogs, goats, sheep, cows, pigs, horses, monkeys or humans, preferably including humans or rats. The administration subject is optionally a subject with long-term and trace cadmium intake without human intervention, such as a human or an animal; or an artificially constructed cadmium-exposed animal model.

[0055] In some alternative embodiments, the administration subject of the preparation for preventing or treating cadmium exposure is a chronic cadmium-exposed rat model, and the chronic cadmium-exposed rat model is a rat with a CdCl2 injection dose of 0.1-0.4 mg / kg body weight, such as, but not limited to, a rat with a CdCl2 injection dose of 0.1, 0.2, 0.3 or 0.4 mg / kg body weight. The injection is preferably intraperitoneal injection. For this chronic cadmium-exposed rat model, calculated based on the amount of pine pollen in the ingested preparation, the daily intake dose of the preparation for preventing or treating cadmium exposure is 0.2-2 g / kg body weight, such as, but not limited to, 0.2, 0.4, 0.5, 0.6, 0.8, 1.0, 1.2, 1.5, 1.6, 1.8 or 2.0 g / kg body weight, and the ranges between any two of the foregoing. For example, it can be, but not limited to, 0.2-0.6 g / kg body weight, 0.6-2.0 g / kg body weight, 1-2.0 g / kg body weight, or 0.5-1.5 g / kg body weight.

[0056] In some alternative embodiments, the administration subject of the preparation for preventing or treating cadmium exposure is a human, and the clinically adult daily dosage range deduced from the effective dose for preventing or treating the chronic cadmium-exposed rat model is 1.2-12 g / kg body weight (calculated based on the amount of pine pollen in the ingested preparation), such as, but not limited to, 1.2, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 g / kg body weight, and the ranges between any two of the foregoing.

[0057] The preparation for preventing or treating cadmium exposure includes a single-agent preparation or a compound preparation. In addition to containing pine pollen, the compound preparation may optionally contain other acceptable active ingredients or auxiliary active ingredients in the art for preventing, alleviating or treating cadmium exposure. The dosage forms of the preparation for preventing or treating cadmium exposure include, but are not limited to, capsules, tablets, granules, pills, dripping pills, syrups, oral liquids. The preparation for preventing or treating cadmium exposure is also optionally a sustained-release preparation or a controlled-release preparation, such as a sustained-release tablet, a sustained-release capsule, a controlled-release tablet or a controlled-release capsule.

[0058] Based on the above experimental findings, the present invention also provides a method for promoting cadmium excretion in the body for non-diagnostic and non-therapeutic purposes, which includes administering pine pollen to the body or administering the above-mentioned preparation for preventing or treating cadmium exposure. The promotion of cadmium excretion in the body for non-diagnostic and non-therapeutic purposes includes, but is not limited to, promoting cadmium excretion in a cadmium-exposed animal model to study drugs related to cadmium exposure or physiological and biochemical functions related to cadmium exposure.

[0059] In some alternative embodiments, the method for promoting cadmium excretion in the body for non-diagnostic and non-therapeutic purposes includes constructing a cadmium-exposed animal model and feeding pine pollen to the cadmium-exposed animal model. The construction of the cadmium-exposed animal model preferably includes constructing a chronic cadmium-exposed rat model, and the construction of the chronic cadmium-exposed rat model includes injecting rats with CdCl2 at a dose of 0.1 - 0.4 mg / kg body weight, for example, but not limited to, 0.1, 0.2, 0.3, or 0.4 mg / kg body weight. The preferred daily intake dose of pine pollen for the cadmium-exposed model rats prepared by the above preparation method is 0.2 - 2 g / kg body weight, for example, but not limited to, 0.2, 0.4, 0.5, 0.6, 0.8, 1.0, 1.2, 1.5, 1.6, 1.8, or 2.0 g / kg body weight, as well as the ranges between any two of the above. For example, but not limited to, 0.2 - 0.6 g / kg body weight, 0.6 - 2.0 g / kg body weight, 1 - 2.0 g / kg body weight, or 0.5 - 1.5 g / kg body weight.

[0060] The technical solutions and beneficial effects of the present invention will be further described below in conjunction with preferred embodiments.

[0061] Experimental materials and instruments:

[0062] Experimental materials: Pine pollen, the pollen of Pinaceae plants, a powdery substance obtained after sand removal, impurity removal, and wall breaking, provided by Yantai New Era Health Industry Co., Ltd.; anhydrous CdCl2, hematoxylin and eosin staining solution, ethanol, paraffin, xylene (purchased from Beijing Chemical Reagent Company); glutathione peroxidase GSH-Px detection kit, malondialdehyde MDA detection kit (purchased from Nanjing Jiancheng Bioengineering Institute).

[0063] Experimental instruments: Atomic absorption spectrophotometer (HITACHI ZA3000), microplate reader (TECAN InfiniteProM200), balance (0.0001 g, Sartorius), microtome (Leica RM2235), embedding machine (Leica EG1150H), coverslipper (Leica CV5030), microscope (Leica DM2000).

[0064] Experimental animals: SPF-grade adult male SD rats were purchased from Beijing Spey Foster Biotechnology Co., Ltd., and the animal license number was SCXK(Jing)2019-0010.

[0065] The experimental grouping and the method for establishing a chronic cadmium-exposed rat model are as follows:

[0066] A chronic cadmium-exposed rat model was established by intraperitoneal injection of a low-dose CdCl2 solution. The specific method was as follows: 50 adult male SD rats weighing 180-220 g were randomly divided into 5 groups, namely the control group (Con), the cadmium-exposed model group (Cd-E), the low-dose pine pollen intervention group (PP-L, 0.2 g / kg bw), the medium-dose pine pollen intervention group (PP-M, 0.4 g / kg bw), and the high-dose pine pollen intervention group (PP-H, 0.6 g / kg bw). The model group and the pine pollen intervention groups were intraperitoneally injected with CdCl2 at a concentration of 0.2 mg / kg bw (calculated as Cd), the blank control group was injected with normal saline and fed with maintenance feed; the model group was injected with CdCl2 and fed with maintenance feed; the pine pollen intervention groups were injected with CdCl2 and fed with pine pollen.

[0067] Example 1

[0068] Effect of pine pollen on the body weight of chronic cadmium-exposed rats:

[0069] According to the above experimental grouping and model establishment method, the rats were conventionally raised and the model was established. The body weight of the rats was measured every day, the data were recorded, the average daily body weight of each group of rats was calculated, and the body weight change curve was plotted. The body weight change curve of the rats is as Figure 1 shown: The body weight of the rats in the cadmium-exposed model group and the body weight of the rats intervened with different doses of pine pollen were comparable to that of the normal control group rats, with no obvious differences. The body weights of the rats in each group were in a stable growth state during the whole experiment, with the same growth trend and similar values, indicating that chronic cadmium exposure had no obvious effect on body weight and did not cause a sharp increase or decrease in the body weight of the rats.

[0070] Example 2

[0071] Effect of pine pollen on the antioxidant capacity of chronic cadmium-exposed rats:

[0072] According to the above experimental grouping and model establishment method, the rats were conventionally raised and the model was established. 12 hours before the end of the experiment, all the rats were fasted but not water-deprived. At the end of the experiment, the rats were anesthetized with chloral hydrate, and blood was taken from the femoral artery to measure the activity of glutathione peroxidase and the content of lipid peroxide MDA. The two indexes were detected using a kit.

[0073] The changes in the activity of antioxidant enzymes and the content of peroxidation products in the blood of rats after pine pollen intervention in cadmium-exposed rats are shown in the appendix Figure 2A andFigure 2B as shown Figure 2A Regarding the effect on the content of antioxidant enzymes, pine pollen intervention can increase the activity of glutathione peroxidase; Figure 2B Regarding the effect on the lipid peroxidation product MDA, pine pollen intervention can significantly inhibit the production of MDA, especially in the high-dose pine pollen group, which has a significant difference compared with the model group (p<0.05).

[0074] Example 3

[0075] Effect of pine pollen on fecal cadmium and urinary cadmium in rats with chronic cadmium exposure:

[0076] Rats were conventionally raised, subjected to cadmium exposure modeling, and pine pollen intervention according to the above experimental grouping and model establishment methods. The feces and urine of rats in each group were collected every day, the wet weight of feces and urine volume were measured and recorded, the feces and urine collected every 7 days were mixed, sampled, and the cadmium content was measured. Cadmium was determined by atomic absorption spectrometry in GB 5009.15-2014.

[0077] The changes in cadmium content in feces and urine of rats after pine pollen intervention for cadmium exposure are shown in Appendix Figure 3 and Appendix Figure 4 as shown. It can be seen from the figure that after intraperitoneal injection of CdCl2 (0.2 mg / kg bw, calculated as Cd) to rats, the cadmium content in feces and urine of the rat cadmium exposure model group and the pine pollen intervention group increased very significantly, with extremely significant differences compared with the normal group (p<0.1). After pine pollen intervention, the cadmium excretion in feces and urine of rats increased significantly compared with the model group, especially in the medium- and high-dose pine pollen groups, with extremely significant differences compared with the model group (p<0.1). The cadmium excretion effect of pine pollen shows a certain positive correlation with the intake.

[0078] The effect of pine pollen on the total excretion of fecal cadmium and urinary cadmium in cadmium-exposed rats is shown in the data of Table 1: Cadmium element is mainly excreted from urine, and the excretion amount accounts for 90-92% of the total cadmium excretion. There are significant differences in the excretion amounts of fecal cadmium and urinary cadmium in the high- and medium-dose pine pollen groups compared with the model group (p<0.05).

[0079] Table 1. Effect of pine pollen on the total excretion of fecal cadmium and urinary cadmium in cadmium-exposed rats

[0080]

[0081] Example 4

[0082] Effect of pine pollen on cadmium in the liver, kidney, testis and femur of rats with chronic cadmium exposure:

[0083] Rats were conventionally raised, subjected to cadmium exposure for model establishment, and intervened with pine pollen according to the above experimental grouping and model establishment method. Twelve hours before the end of the experiment, all rats were fasted but allowed to drink water. At the end of the experiment, after anesthesia and sacrifice with chloral hydrate, tissues such as the liver, kidney, bone, and testis were collected. Cadmium was determined by atomic absorption spectrometry in GB 5009.15-2014.

[0084] The results of the changes in cadmium content in the liver, kidney, testis, and femur of rats after pine pollen intervention for cadmium-exposed rats are as Figure 5A , Figure 5B , Figure 5C and Figure 5D shown. The experimental results show that the organs most prone to cadmium accumulation after cadmium poisoning in rats are the kidney, testis, and liver. Pine pollen has a very obvious cadmium-excreting effect and shows a dose-effect. The high-dose pine pollen group has an extremely significant effect on cadmium excretion. The cadmium content in the kidney, testis, and liver decreased by 89%, 96%, and 92% respectively compared with the model group (p<0.01), and the cadmium content in the femur decreased significantly, by 65% compared with the model group (p<0.01). The cadmium content in the kidney, testis, liver, and femur of rats in the medium-dose pine pollen group and the low-dose pine pollen group decreased significantly compared with the model group, and there were significant differences (p<0.05).

[0085] Example 5

[0086] Effect of pine pollen on the morphology of the kidney and testis of rats with chronic cadmium exposure:

[0087] Rats were conventionally raised, subjected to cadmium exposure for model establishment, and intervened with pine pollen according to the above experimental grouping and model establishment method. Twelve hours before the end of the experiment, all rats were fasted but allowed to drink water. At the end of the experiment, after anesthesia and sacrifice with chloral hydrate, the kidney and testis tissues were quickly removed under low-temperature conditions, the surface blood was rinsed with low-temperature normal saline, blotted dry with filter paper, and fixed in 10% formaldehyde solution for trimming. The preparation of the pre-slice for HE staining was carried out according to the following steps: dehydration with ethanol step by step, transparency treatment with xylene, wax immersion, dewaxing, debenzolization, HE staining, dehydration, transparency, and mounting. Photographs were taken with an optical microscope (200×).

[0088] The results of the effect of pine pollen intervention on the morphology of the kidney and testis of cadmium-exposed rats are as shown in Appendix Figure 6 .

[0089] Figure 6In [description], A is a testicular section of a rat. A1 is the normal group, with the testicular tissue structure being basically normal, the seminiferous tubules having a complete structure, and the red arrows indicating spermatogonia, with the quantity not showing a decrease. A2 is the model group, with the testicular tissue structure being severely abnormal. In the field of view, various spermatocytes are absent in some seminiferous tubules, the remaining spermatogonia are arranged disorderly, and the quantity is significantly reduced. A large amount of protein mucus can be seen in the lumen. A3 is the high-dose pine pollen group, with the testicular tissue structure being mildly abnormal. In the field of view, various spermatocytes are absent in the seminiferous tubules, the quantity is significantly reduced, and residual spermatids can be seen in the lumen. A4 is the medium-dose pine pollen group, with the testicular tissue structure being mildly abnormal. In the field of view, the cells in some seminiferous tubules are arranged loosely, and the quantity does not show an obvious decrease. A5 is the low-dose pine pollen group, with the testicular tissue structure being severely abnormal, and all interstitial cells and various spermatocytes being necrotic, with the cell nuclei being pyknosed and deeply stained.

[0090] Figure 6 In [description], B is a photograph of a kidney tissue section of a rat. B1 is the normal group, with the kidney tissue structure being normal, the glomeruli having a complete structure, and the glomerular capsule cavity not showing dilation; the renal tubular epithelial cells are arranged regularly; the interstitial space does not show dilation; no pathological changes such as inflammation, edema, and fibrosis are seen in the tissue. B2 is the cadmium-exposed model group, with the kidney tissue structure being moderately abnormal, the glomeruli showing obvious lobulation; some renal tubular epithelial cells are edematous and vacuolated, and some brush borders are shed. B3 is the high-dose pine pollen group, with the kidney tissue structure being mildly abnormal, the glomeruli showing lobulation, and protein mucus exuding into the capsule cavity. Protein mucus can be seen in some renal tubules. B4 is the medium-dose pine pollen group, with the kidney tissue structure being basically normal, the glomeruli having a complete structure, and the glomerular capsule cavity not showing dilation; some brush borders can be seen shed in some renal tubules. B5 is the low-dose pine pollen group, with the kidney tissue structure being basically normal, the glomerular capsule cavity not showing dilation, and slight pathological changes such as inflammation, edema, and fibrosis in the tissue.

[0091] Example 6

[0092] Effect of pine pollen on zinc in the liver, kidney, testis, and femur of rats with chronic cadmium exposure:

[0093] The experimental operation was the same as in Example 4, and zinc was determined by the flame atomic absorption method in GB 5009.14-2017. The results of the changes in zinc content in the liver, kidney, testis, and femur of rats after pine pollen intervention in cadmium-exposed rats are shown in Figure 7A , Figure 7B , Figure 7C and Figure 7D . The experimental results show that cadmium exposure in rats has a certain effect on the zinc content in the organs of rats, especially on kidney zinc and testis zinc (p<0.05). Cadmium exposure can cause zinc loss in the kidneys and testes of rats, and pine pollen intervention has an obvious recovery effect on the zinc content in the kidneys and testes of cadmium-exposed rats, and is close to the control group level.

[0094] Example 7

[0095] Effect of Pine Pollen on Zinc in Feces and Urine of Rats Exposed to Chronic Cadmium

[0096] The experimental operation was the same as that in Example 3. Zinc was determined by flame atomic absorption spectrometry in GB 5009.14-2017. After pine pollen intervened in cadmium-exposed rats, the changes in zinc in the feces and urine of the rats were shown in Appendix Figure 8A and Appendix Figure 8B as shown. The experimental results showed that the excretion of zinc in the feces and urine of the rats increased after pine pollen intervention, but there was no significant difference. Combining with the fact that the zinc content in the organs of the rats could basically maintain at the control group level after pine pollen intervention in Example 6, it could be inferred that pine pollen intervention would not promote the excretion of zinc in the rat body.

[0097] Finally, it should be noted that: the above embodiments are only used to illustrate the technical solutions of the present invention, rather than limiting it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of pine pollen in the preparation of a preparation for preventing or treating chronic cadmium exposure, wherein the administration object of the preparation is a mammal; the preparation for preventing or treating cadmium exposure is a capsule, tablet, granule, pill, freeze-dried powder, syrup or oral liquid.

2. The application according to claim 1, characterized in that The pill includes a dripping pill.

3. The application according to claim 1, characterized in that, The mammal is a human, rat, mouse, guinea pig, hamster, rabbit, ferret, cat, dog, goat, sheep, cow, pig, horse or monkey.

4. The application according to claim 1, characterized in that, The preparation for preventing or treating cadmium exposure is a single-agent preparation or a compound preparation.

5. The application according to claim 1, wherein The preparation for preventing or treating cadmium exposure is a sustained-release preparation or a controlled-release preparation.

6. Use of pine pollen in the preparation of a preparation for promoting cadmium excretion from the body, wherein the administration object of the preparation is a mammal; the preparation for promoting cadmium excretion from the body is a capsule, tablet, granule, pill, freeze-dried powder, syrup or oral liquid.

7. The application according to claim 6, wherein The pill includes a dripping pill.

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