Traditional Chinese medicine charcoal medicine with lipid metabolism regulating effect

Abstract

The invention belongs to the technical field of medicines, and particularly relates to preparation and application of cacumen biotae charcoal. Animal pharmacological experiments and cell pharmacological experiments find that the cacumen biotae charcoal has a definite lipid-lowering effect on mice in a physiological state and acute and chronic hyperlipidemia animal models.

Description

Technical Field

[0001] The invention relates to the field of medicine, and in particular to application of Platycladus orientalis leaf charcoal. Background Art

[0002] Lipids are important substances for storing and providing energy in the human body, and are also important structural components of biological membranes. Lipid metabolism is a very important and complex biochemical reaction in the human body. It mainly refers to the process of digestion, absorption, synthesis and decomposition of fat in the human body under the catalysis of various related enzymes to ensure the normal operation of physiological functions. Therefore, maintaining lipid metabolic homeostasis is of great significance for life activities. Abnormal lipid metabolism may cause a variety of diseases, such as hyperlipidemia, hypertension, coronary artery atherosclerosis, fatty liver, and even myocardial infarction, stroke and other diseases that seriously endanger human health.

[0003] Platycladus orientalis leaves are the dried branches and leaves of Platycladus orientalis Franco., a plant of the Cupressaceae family. It has a fresh fragrance and a bitter, slightly pungent taste. It has the effects of cooling blood and stopping bleeding, resolving phlegm and relieving cough, promoting dampness and stopping leukorrhea, dispelling wind and stopping itching, and dispelling wind and removing dampness. It is clinically used for various bleeding syndromes such as vomiting blood due to blood heat, epistaxis, hematuria, bloody dysentery, metrorrhagia, cough due to lung heat, thick phlegm that is difficult to cough up, hair loss due to blood heat, and premature graying of hair. It is also used for red and white leukorrhea, leprosy, rheumatism and other pains caused by damp heat. Platycladus orientalis leaf charcoal is a traditional Chinese medicine made from Platycladus orientalis leaves, which has the effect of cooling blood and stopping bleeding. Summary of the invention

[0004] The use of Platycladus orientalis leaf charcoal in preparing a lipid-lowering composition. The metabolic disease is any metabolic disease with lipid accumulation as the main pathological change, such as hyperlipidemia (any one of hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, and low high-density lipoprotein cholesterol), fatty liver, etc.

[0005] The metabolic disease is hyperlipidemia.

[0006] The composition may or may not contain pharmaceutically acceptable excipients.

[0007] The composition is any one of a medicine, a health product, and a food, and the composition can be prepared into a clinically acceptable oral preparation; further preferably, the clinically acceptable oral preparation is one or more of a pill, a capsule, a tablet, a granule or a liquid oral preparation; more preferably, the oral preparation is a granule.

[0008] Specifically, the technical solution of the present invention is as follows.

[0009] The present invention relates to the biological activity of Platycladus orientalis leaf charcoal and its application in the field of medicine. Platycladus orientalis leaf charcoal has the effect of regulating lipid metabolism in mice under normal nutritional status, acute hyperlipidemia model mice induced by egg yolk emulsion, and chronic hyperlipidemia model rats fed with high-fat diet, showing lipid-lowering biological activity. It is mainly used for treating hyperlipidemia and regulating lipid metabolism homeostasis activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1 .Effects of Platycladus orientalis charcoal on lipids in mice under normal nutritional status. The animals were divided into 4 groups: control group (Control), Platycladus orientalis charcoal administration groups with different doses [high (H-PCC-CDs, 1.64 mg / kg), medium (M-PCC-CDs, 0.96 mg / kg), low (L-PCC-CDs, 0.48 mg / kg)]. Figure 1 a: Experimental flow chart; Figure 1 b: Mouse weight; Figure 1 c: Food intake of mice; Figure 1 d: mouse plasma TC; Figure 1 e: mouse plasma TC; Figure 1 f: mouse plasma FFA; Figure 1 g: mouse plasma LDL-C; Figure 1 h: mouse plasma HDL-C; Figure 1 i: Photo of mouse liver; Figure 1 j: liver / body weight ratio of mice; Figure 1 k: TC in mouse liver tissue; Figure 1 l: TG in mouse liver tissue; Figure 1 m: ALT in mouse blood; Figure 1 n: AST in mouse blood; Figure 1 o: H&E staining results of mouse liver tissue.

[0011] Figure 2 .Effect of Platycladus orientalis charcoal on elevated blood lipids in acute hyperlipidemia mouse model induced by a single intraperitoneal injection of 75% EYE. The animals were divided into 5 groups: control group (Control), acute hyperlipidemia model group induced by egg yolk emulsion (EYE), and different doses of Platycladus orientalis charcoal administration groups [1.64 mg / kg, 0.96 mg / kg, 0.48 mg / kg]. Figure 2 a: Experimental flow chart; Figure 2 b: Mouse weight; Figure 2 d: mouse plasma TC; Figure 2 e: mouse plasma TC; Figure 2 f: mouse plasma LDL-C; Figure 2 g: mouse plasma HDL-C (20h); Figure 2 h: mouse plasma HDL-C (48h); Figure 2i: liver / body weight ratio of mice; Figure 2 j: TC in mouse liver tissue; Figure 2 k: TG in mouse liver tissue.

[0012] Figure 3 .Study on the effect of Platycladus orientalis charcoal on lipid-lowering effect in rat model of chronic hyperlipidemia fed with high-fat diet. The animals were divided into 5 groups: normal control group (Control), hyperlipidemia model group fed with high-fat diet (HFD), and Platycladus orientalis charcoal administration groups with different doses [1.64mg / kg, 0.96mg / kg, 0.48mg / kg]. Figure 3 a: Experimental flow chart; Figure 3 b: Appearance of mice; Figure 3 c: Mouse weight; Figure 3 d: mouse plasma TC; Figure 3 e: mouse plasma TC; Figure 3 f: TC in mouse liver tissue; Figure 3 g: TG in mouse liver tissue; Figure 3 h: mouse plasma LDL-C; Figure 3 i: mouse plasma HDL-C; Figure 3 j: Photo of mouse liver; Figure 3 k: liver / body weight ratio of mice; Figure 3 l: H&E staining results of mouse liver tissue.

[0013] Figure 4 .Effect of Platycladus orientalis charcoal on lipid metabolism-related proteins (detected by Western blotting). The animals were divided into 5 groups: normal control group (Control), high-fat diet-fed hyperlipidemia model group (HFD), and Platycladus orientalis charcoal administration groups of different doses [1.64 mg / kg, 0.96 mg / kg, 0.48 mg / kg]. DETAILED DESCRIPTION

[0014] The technical solution of the present invention will be clearly and completely described below in conjunction with specific embodiments, but the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. All other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0015] Example 1: Preparation of Platycladus orientalis leaf charcoal extract

[0016] Take an appropriate amount of Platycladus orientalis leaf charcoal, crush it, add water and boil it for 3 times, each time for 1 hour, filter it with gauze first, then filter it with a 0.22μm microporous filter membrane, and concentrate the filtrate to obtain the solution that is the Platycladus orientalis leaf charcoal extract, recorded as PCC-CDs.

[0017] Example 2: Study on the lipid-lowering effect of Platycladus orientalis charcoal on mice under normal nutritional status

[0018] 1. Experimental Animals

[0019] In this experiment, clean-grade Kunming male mice (7 weeks old, weighing 30 g ± 2 g) were used as experimental subjects. They were kept in a standard environment with free access to water, given a standard maintenance diet, and a 12-h light cycle, and were adaptively raised for one week.

[0020] 2. Drug administration, modeling and sampling of experimental animals

[0021] The animals were weighed and divided into 4 groups according to a random number table: blank control group (Control), high-, medium-, and low-dose administration groups of Platycladus orientalis charcoal (H-PCC-CDs, 1.64 mg / kg; M-PCC-CDs, 0.96 mg / kg; L-PCC-CDs, 0.48 mg / kg), with 8 mice in each group. Each group was intragastrically administered with a dose of 0.1 mL / 10 g per day, and the weight changes of the mice were recorded. The experimental period was 5 weeks.

[0022] After the last administration, the mice were anesthetized and their body shape and hair were observed and photographed. The mice were killed by dislocating the cervical vertebra after removing the eyeballs and taking blood. The mouse liver was removed, and the levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL-C), high-density lipoprotein (HDL-C), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) in serum were detected using an automatic biochemical analyzer. The supernatant of liver tissue homogenate was used to determine the TC and TG content in liver tissue (Nanjing Jiancheng Biochemical Kit).

[0023] 3 Effects of Platycladus orientalis leaf charcoal on lipid content in mice under normal nutritional status

[0024] Figure 1 The effects of Platycladus orientalis leaf charcoal on lipid content in plasma and liver tissue of mice under normal nutritional status were demonstrated. Figure 1 a is the experimental flow chart. The results showed that the physiological parameters of mice, such as body weight and food intake, remained stable without significant changes ( Figure 1 bc). In addition, compared with the blank control group, intervention with Platycladus orientalis charcoal caused a decrease in the levels of plasma cholesterol, triglycerides, and FFA in mice ( Figure 1 df). In addition, the mice's low-density lipoprotein cholesterol and high-density lipoprotein cholesterol also decreased ( Figure 1 gh). Considering that liver tissue is a central regulator of lipid homeostasis, we observed no significant changes in the structure and shape of the liver, but the liver tissue treated with Platycladus orientalis charcoal showed a lighter reddish brown color compared to the liver tissue of healthy mice. In addition, the liver tissue homogenate of the mice treated with Platycladus orientalis charcoal was clearer and also significantly different ( Figure 1i). Notably, Platycladus orientalis charcoal reduced liver cholesterol and triglyceride levels, while the ratio of liver weight to body weight did not change significantly ( Figure 1 jl). The results of H&E staining showed no obvious pathological changes ( Figure 1 In conclusion, these results suggest that Platycladus orientalis charcoal can reduce lipid content to below normal levels in mice under normal nutritional conditions and affect the activities of hepatic aminotransferases without causing obvious morphological damage.

[0025] Example 3: Study on the lipid-lowering effect of Platycladus orientalis charcoal on acute hyperlipidemia model mice induced by 75% egg yolk emulsion

[0026] 1. Experimental Animals

[0027] In this experiment, clean-grade Kunming male mice (7 weeks old, weighing 30 g ± 2 g) were used as experimental subjects. They were kept in a standard environment with free access to water, given a standard maintenance diet, a 12-h light cycle, and adaptively raised for 7 days.

[0028] 2. Drug administration, modeling and sampling of experimental animals

[0029] Preparation of 75% egg yolk emulsion: Take 30 mL of fresh egg yolk and add it to 10 mL of normal saline and mix thoroughly.

[0030] The experimental animals were randomly divided into a normal control group (Control), an acute hyperlipidemia model group induced by egg yolk emulsion (EYE), and a high-dose (EYE+H-Oriental Platycladus orientalis charcoal, 1.64 mg / kg), a medium-dose (EYE+M-Oriental Platycladus orientalis charcoal, 0.96 g / mL), and a low-dose (EYE+L-Oriental Platycladus orientalis charcoal, 0.48 g / mL) administration group of Platycladus orientalis charcoal. The administration was given by intragastric administration every day, and the experimental period was 5 weeks. After the last intragastric administration, each mouse was intraperitoneally injected with 75% egg yolk emulsion at a dose of 0.1 mL / 10 g. After 20 h and 48 h of intraperitoneal injection of egg yolk emulsion, the mice were eyeballed and blood was collected, and the mice were killed by cervical dislocation. They were fasted but not watered for 12 h before blood collection. The mouse serum TC, TG, LDL-C, and HDL-C were detected using a fully automatic biochemical analyzer.

[0031] 3. Effect of Platycladus orientalis charcoal on lipid content in acute hyperlipidemia model mice

[0032] To evaluate the effect of Platycladus orientalis charcoal on lipid overaccumulation in pathological models, we prepared a 75% egg yolk emulsion (EYE) from food source egg yolk in sterile saline. Figure 2 a). Figure 2 As shown in b, there was no significant difference in weight gain among the five groups of mice. In addition, the plasma of the mice in the group receiving orientalis charcoal appeared transparent and bright, while the plasma of the mice receiving EYE intervention showed a slightly milky and turbid appearance ( Figure 2 c). In addition, administration of Platycladus orientalis charcoal resulted in a decrease in plasma cholesterol, triglycerides, and low-density lipoprotein levels, which were increased after EYE intervention ( Figure 2 df). Figure 2 As shown in Figure gh, EYE caused a temporary increase in plasma HDLC after 20 hours of intervention, which decreased after 48 hours, with no significant difference compared with the control group. However, 48 hours after EYE injection, the plasma HDLC level in the Platycladus orientalis charcoal-administered group decreased to below normal levels. Interestingly, there were no significant changes in liver cholesterol and triglyceride contents after EYE intervention. In contrast, the administration of Platycladus orientalis charcoal led to a further decrease in liver cholesterol and triglyceride contents compared with the blank control group, while no significant difference was observed in the ratio of liver weight to body weight ( Figure 2 ik). Collectively, these data suggest that T. orientalis charcoal alleviates the transient plasma accumulation induced by exogenous lipid overload.

[0033] Example 4: Study on the lipid-lowering effect of Platycladus orientalis leaf charcoal on hyperlipidemia model rats fed with a high-fat diet

[0034] 1. Experimental Animals

[0035] Clean-grade Sprague Dawley male rats (5 weeks old, weighing 160 g ± 10 g) were used as experimental subjects in this experiment. They were kept in a standard environment with free access to water, given a standard maintenance diet, a 12-h light cycle, and adaptively raised for 7 days.

[0036] 2. Drug administration, modeling and sampling of experimental animals

[0037] The experimental animals were randomly divided into a normal control group (Control), a hyperlipidemia model group fed with a high-fat diet (HFD), and a high-dose (HFD+H-Platifolia charcoal, 1.64 g / mL), a medium-dose (HFD+M-Platifolia charcoal, 0.96 g / mL), and a low-dose (HFD+L-Platifolia charcoal, 0.48 g / mL) administration group of Platycladus orientalis leaf charcoal. Except for the normal control group, which was given a standard maintenance diet, each group began to be fed a high-fat diet and was given drugs by gavage every day. The blank group and the model group were given an equal volume of deionized water. The rats were weighed every 7 days, and the experimental period was 7 weeks.

[0038] After the experiment, the rats were fasted but not watered for 12 hours. After anesthesia, the rats were observed for body shape, hair, etc. and photographed. Blood was taken from the abdominal aorta and the rats were killed by cervical dislocation. The rat livers were removed, and the liver tissue pathology and hepatocyte lipid deposition were observed by H&E staining and oil red staining. The serum TC, TG, LDL-C, and HDL-C levels were detected by an automatic biochemical analyzer; the TC and TG contents in the liver tissue were determined by the supernatant of 10% liver tissue homogenate.

[0039] 3. Effect of Platycladus orientalis charcoal on lipid content in hyperlipidemia model rats

[0040] Next, we investigated whether Platycladus orientalis charcoal could improve hepatic steatosis in HFD-fed rats ( Figure 3 a). Figure 3 As shown in bc, after 7 weeks of treatment with Platycladus orientalis charcoal, the body weight of rats decreased by 14% compared with the blank group. In addition, Platycladus orientalis charcoal treatment significantly reduced the plasma cholesterol and triglyceride levels of rats fed with HFD by about 1.5 times ( Figure 3 de), which is consistent with the results of lowering liver cholesterol and triglycerides ( Figure 3 fg). Notably, Platycladus orientalis charcoal also reduced the accumulation of low-density lipoprotein cholesterol, while high-density lipoprotein cholesterol in plasma did not change significantly ( Figure 3 hi). The liver tissue of rats fed with HFD was diffusely enlarged, with blunt and thick edges, smooth surface, lighter color, and greasy feeling. After intervention with Platycladus orientalis charcoal, the swelling of the liver tissue was not obvious, and the color turned ruddy ( Figure 3 j). In the model control group fed with a high-fat diet, the liver coefficient increased significantly, but after intervention with Platycladus orientalis charcoal, the liver coefficient decreased significantly ( Figure 3 k). In addition, we performed HE staining on the liver tissues of each group. The model control group rats fed with a high-fat diet showed vacuoles of varying sizes in the hepatic cytoplasm, showing diffuse macrovesicular fatty changes. In the oil red staining, the model control group also showed a large number of red lipid droplets deposited, and lipid infiltration was obvious. After the intervention of Platycladus orientalis leaf charcoal, these pathological changes were significantly alleviated. ( Figure 3 l). These results indicate that Platycladus orientalis leaf charcoal can effectively improve obesity caused by unbalanced diet and alleviate the pathological process of fatty degeneration of the liver.

[0041] 4. Effects on the expression of lipid metabolism-related proteins in hyperlipidemia model rats

[0042] Western blot was used to detect the expression of proteins related to cholesterol and fatty acid synthesis and fatty acid β-oxidation in rat liver tissue. Figure 4, it can be seen that p-AMPKα activity was significantly decreased in HFD-fed mice, while it was greatly restored in PCC-CDss. There was no significant change in endogenous AMPKα. The accumulation of cleaved ACC in the mouse liver was significantly reduced by PCC-CDss. PCC-CDss also reduced the amount of mature SREBP-1c, which increased 1.9-fold in HFD-fed mice. In addition, hyperlipidemia increased FASN protein 1.6-fold, while PCC-CDss reduced it to near normal levels, which may be related to the reduction of SREBP-1c. These results suggest that PCC-CDss reduces new lipogenesis by inhibiting SREBP-1c and activating the AMPK pathway, resulting in a decrease in ACC and FAS activity. In addition, we observed that PCC-CDss increased the protein expression of carnitine palmitoyltransferase I (CPT1α), an enzyme that shuttles fatty acid esters into mitochondria.

[0043] AMPK delays the aging process of cells and organisms through multiple molecular mechanisms such as regulating energy metabolism, reducing oxidative stress, reducing protein synthesis, and increasing DNA repair, thereby prolonging life. Pleurotus eryngii charcoal significantly affects the activation of AMPK, so it has a positive effect on delaying aging and longevity.

Claims

1. Use of Platycladus orientalis leaf charcoal in preparing a composition for lipid metabolic diseases.

2. The lipid metabolism disorder according to claim 1, characterized in that The metabolic disease may be hyperlipidemia (or any one of hypercholesterolemia, hypertriglyceridemia, mixed hyperlipidemia, and low high-density lipoprotein cholesterol), fatty liver, and obesity.

3. The use for treating lipid metabolism disorders according to claim 1, characterized in that: The metabolic diseases are mainly caused by lipid metabolism disorders and are a type of disease with excessive lipid accumulation as the main pathological change.

4. The method for regulating lipid metabolism homeostasis according to claim 1, wherein: The composition may or may not contain pharmaceutically acceptable excipients.

5. The use for treating lipid metabolism disorders according to claim 1, characterized in that: The arborvitae leaf charcoal and pharmaceutically acceptable excipients are prepared into clinical preparations; the clinical preparations include but are not limited to injections, oral liquid preparations, hard capsules, soft capsules, tablets, suppositories, water pills, honey pills, concentrated pills, ointments, powders, emulsions, pastes, and smears.

6. The use according to claim 1, characterized in that: Use with other drugs or alone.

7. The method for regulating lipid metabolism homeostasis according to claim 1, wherein: The composition is any one of a medicine, a health product, and a food, and the composition can be prepared into a clinically acceptable oral preparation; further preferably, the clinically acceptable oral preparation is one or more of a pill, a capsule, a tablet, a granule or a liquid oral preparation; more preferably, the oral preparation is a granule.

8. Use of Platycladus orientalis leaf charcoal in preparing medicines for treating or alleviating hyperlipidemia and obesity.

9. Use of Platycladus orientalis leaf charcoal in anti-aging and longevity promoting medicines.

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