Application of cinnamyl aldehyde

Abstract

The invention discloses application of cinnamyl aldehyde, and cinnamyl aldehyde can regulate synthesis and secretion of androgen testosterone and can be applied to related products for regulating synthesis and secretion of androgen testosterone. In addition, tests prove that cinnamyl aldehyde increases synthesis and secretion of testicular interstitial cell androgen by improving activity of a steroid hormone synthesis nuclear receptor promoter and up-regulating expression of steroid hormone synthetase.

Description

Technical Field

[0001] The invention belongs to the technical field of life sciences, and particularly relates to an application of cinnamaldehyde. Background Art

[0002] Cinnamon, belonging to the genus Cinnamomum and the Lauraceae family, has been a popular spice around the world for thousands of years. In my country, it is abundant and cultivated across vast tracts, primarily in the southwest and southeast regions. In Traditional Chinese Medicine, cinnamon is often used to tonify heat and yang, dissipate heat and relieve pain, and warm the meridians.

[0003] Testosterone is the primary hormone in male animals that maintains reproductive differentiation, secondary sexual characteristics, auxiliary gonadal development, and spermatogenesis. Testosterone is produced and secreted by Leydig cells in the testes, with multiple signaling pathways and enzymes involved in this process. Numerous studies have demonstrated that cinnamon has antibacterial, antiviral, antifungal, antioxidant, antitumor, antihypertensive, lipid-lowering, hypoglycemic, gastric protective, and immune-enhancing effects. Currently, there are no reports on the role of cinnamaldehyde in regulating androgen synthesis and secretion. Summary of the Invention

[0004] In view of this, in a first aspect, the present invention provides a culture method for increasing the expression of CYP17A1, CYP11A1, and 3β-HSD proteins in mouse Leydig cells and reducing the expression of StAR protein in mouse Leydig cells for non-therapeutic purposes to increase the testosterone secretion of mouse Leydig cells, comprising: treating mouse Leydig cells with cinnamaldehyde.

[0005] In some embodiments, 5×10 -5 Mouse Leydig cells were treated with cinnamaldehyde for 24 hours.

[0006] In a second aspect, the present invention provides a use of cinnamaldehyde to prepare a preparation, which is used to increase the expression of CYP17A1, CYP11A1, and 3β-HSD proteins in mouse Leydig cells, and reduce the expression of StAR protein in mouse Leydig cells to increase testosterone secretion in mouse Leydig cells.

[0007] In some embodiments, the preparation is a reagent, a pharmaceutical agent, etc.; the preparation comprises 5×10 -5 Cinnamaldehyde of M.

[0008] In a third aspect, the present invention provides a use of cinnamaldehyde for non-therapeutic purposes in increasing the expression of CYP17A1, CYP11A1, and 3β-HSD proteins in mouse Leydig cells, reducing the expression of StAR protein in mouse Leydig cells, increasing the activity of steroid synthesis nuclear receptor promoter, and increasing the secretion of testosterone in mouse Leydig cells; the steroid synthesis nuclear receptor promoter includes the SF-1 promoter.

[0009] In a fourth aspect, the present invention provides a use of cinnamaldehyde in the preparation of a functional feed additive for enhancing the expression of 3β-hsd, CYP11A1, and StAR proteins and increasing androgen secretion.

[0010] In some embodiments, the amount of the additive in the feed is 25-75 mg / kg.

[0011] Compared with the prior art, the present invention has the following beneficial effects:

[0012] The present invention has demonstrated through experiments that cinnamaldehyde increases the activity of steroid hormone synthesis nuclear receptor promoters, upregulates the expression of steroid hormone synthase, and increases the synthesis and secretion of androgens in testicular interstitial cells. The invention can be applied to related products for regulating the synthesis and secretion of the androgen testosterone. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 The effect of cinnamaldehyde on testosterone secretion in mouse Leydig cells;

[0014] Figure 2 The effect of cinnamaldehyde on androgen synthase in mouse Leydig cells;

[0015] Figure 3 The effect of cinnamaldehyde treatment on the activity of testosterone secretion nuclear receptor promoter;

[0016] Figure 4 The effects of cinnamaldehyde treatment on body weight, testicular index and serum testosterone concentration in mice;

[0017] Figure 5 The effect of cinnamaldehyde treatment on mouse liver;

[0018] Figure 6 The effect of cinnamaldehyde treatment on mouse testis;

[0019] Figure 7 This is the effect of cinnamaldehyde treatment on the expression of testosterone synthase in mouse testis. DETAILED DESCRIPTION

[0020] To make the objectives, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, 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 based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0021] Example 1: Effects of cinnamaldehyde on Leydig cells in male mice

[0022] Testosterone is the main hormone that maintains the differentiation of reproductive organs, secondary sexual characteristics, the development of accessory glands and spermatogenesis in male animals. Testosterone is synthesized and secreted by Leydig cells, and multiple signaling pathways and enzymes are involved in this process. This example starts from the perspective of cinnamaldehyde regulating testosterone secretion in Leydig cells, and explores its effect on signaling pathways related to steroid hormone synthesis. This example study used methods such as protein immunoblotting, real-time fluorescence quantitative PCR, enzyme-linked immunosorbent assay, flow cytometry and dual luciferase reporter system to obtain the following results:

[0023] (1) Cinnamaldehyde treatment can significantly promote the secretion of testosterone in mouse Leydig cells. After treating mouse Leydig cells with different concentrations of cinnamaldehyde, the testosterone concentration in the cell supernatant was detected, and the results were as shown in the attached figure. Figure 1 As shown. With the increase of cinnamaldehyde treatment time and treatment concentration, it was found that 5×10 -5 Treatment with M's cinnamaldehyde for 24 hours can significantly increase the testosterone secretion of mouse Leydig cells.

[0024] (2) Cinnamaldehyde treatment can significantly increase the expression of androgen synthase in Leydig cells. The experiment used RT-qPCR and WB methods to detect the expression of steroid hormone synthesis-related enzymes in MLTC-1 cells after cinnamaldehyde treatment, as shown in the attached Figure 2 RT-qPCR results showed that 5×10 -5 Cinnamaldehyde treatment of M significantly increased the amount of CYP17A1 mRNA. However, the amount of CYP11A1 and 3β-HSD mRNA decreased significantly. WB test results showed that 5×10 -5 Cinnamaldehyde treatment of M could significantly upregulate the expression of CYP17A1, CYP11A1 and 3β-HSD proteins, while the expression of StAR protein was significantly decreased.

[0025] (3) Cinnamaldehyde treatment can significantly increase the activity of steroid hormone synthesis nuclear receptor promoter. The steroid hormone synthesis nuclear receptor pathway is an important pathway that affects testosterone synthesis. The expression changes of related proteins in the pathway will affect the final testosterone synthesis. The experiment used RT-qPCR and dual luciferase reporter system to analyze the expression of steroid hormone synthesis nuclear receptor and the activity of its promoter. The results are shown in the attached figure. Figure 3 RT-qPCR results showed that 5×10 -5 Cinnamaldehyde treatment of M increased the amount of SF-1 and SHP mRNA and decreased the amount of NUR-77 mRNA. The Nur-77 and SF-1 promoter dual luciferase reporter plasmids were successfully constructed. The dual luciferase reporter system was used to detect 5×10 -5The effects of cinnamaldehyde treatment on the Nur-77 and SF-1 promoter activities were shown in the attached figure. Figure 3 C. 5×10 -5 Cinnamaldehyde treatment of M increased the activity of SF-1 promoter, but had no effect on the activity of Nur-77 promoter.

[0026] Example 2: Effects of feeding cinnamaldehyde-containing feed on male mice

[0027] Through the in vitro experiments in Example 1, it can be found that cinnamaldehyde can regulate the synthesis and secretion of androgens by regulating the activity of the androgen synthesis nuclear receptor promoter and regulating the expression of androgen synthesis-related proteins and enzymes. This example examines the effect of cinnamaldehyde on androgen secretion in male mice through in vivo experiments, thereby providing an experimental basis for the use of cinnamaldehyde or cinnamon as a functional feed additive. This example used methods such as protein immunoblotting, immunohistochemistry, and enzyme-linked immunosorbent assay to obtain the following results:

[0028] (1) Effects of feeding cinnamaldehyde on mice. After feeding mice with feed containing different doses of cinnamaldehyde for 12 days, the changes in body weight, testicular index and serum testosterone concentration of mice in each treatment group were detected. The results are shown in the attached figure. Figure 4 The results showed that feeding 25mg / kg, 50mg / kg and 75mg / kg of cinnamaldehyde could significantly increase the body weight of mice and increase the testosterone concentration in serum, but had no effect on the testicular index of mice.

[0029] (2) Effects of feeding cinnamaldehyde on mouse liver. The effects of feeding different doses of cinnamaldehyde on mouse liver tissue were observed by HE staining. The results are shown in the attached figure. Figure 5 As shown, compared with the PBS plus DMSO group, the liver tissue of mice fed with 25 mg / kg cinnamaldehyde did not show abnormalities. However, the liver tissue of mice fed with 50 mg / kg and 75 mg / kg cinnamaldehyde showed a certain degree of cell swelling and loose cytoplasm.

[0030] (3) Effects of feeding cinnamaldehyde on testicular tissue of mice. The effects of feeding different doses of cinnamaldehyde on testicular tissue of mice were observed by HE staining. The results are shown in the attached figure. Figure 6 As shown, compared with the PBS combined with DMSO group, the number of interstitial cells in the testes of mice in the cinnamaldehyde-fed group was significantly increased, and the increase in interstitial cells in the 25 mg / kg group was the most significant.

[0031] (4) Effect of feeding cinnamaldehyde on the expression of testosterone synthase in mouse testis. Immunohistochemistry and WB were used to detect the expression of enzymes related to testosterone synthesis in testicular tissue. The results are shown in the attached figure. Figure 7As shown. Compared with the PBS plus DMSO group, the expression of 3β-hsd, CYP11A1, and StAR enzymes in the testes of mice fed all cinnamaldehyde groups was significantly increased. Among them, the expression of 3β-hsd enzyme in the testes of mice fed 25 mg / kg cinnamaldehyde was the most significantly increased. The expression of CYP11A1 and StAR enzymes in the testes of mice fed 75 mg / kg cinnamaldehyde was the most significantly increased.

[0032] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. A culture method for increasing the expression of CYP17A1, CYP11A1, and 3β-HSD proteins in mouse Leydig cells for non-therapeutic purposes, reducing the expression of StAR protein in mouse Leydig cells, and increasing the activity of steroidogenic nuclear receptor promoter to increase testosterone secretion in mouse Leydig cells, characterized in that: include: Mouse Leydig cells were treated with cinnamaldehyde.

2. The culture method according to claim 1, wherein 5×10 -5 Mouse Leydig cells were treated with cinnamaldehyde for 24 hours.

3. A method for preparing a preparation using cinnamaldehyde, characterized in that: The preparation is used to increase the expression of CYP17A1, CYP11A1, and 3β-HSD proteins in mouse Leydig cells, reduce the expression of StAR protein in mouse Leydig cells, and enhance the activity of steroidogenic nuclear receptor promoter to increase the secretion of testosterone in mouse Leydig cells.

4. The use according to claim 3, characterized in that The formulation contains 5×10 -5 Cinnamaldehyde of M.

5. A use of cinnamaldehyde for non-therapeutic purposes in increasing the expression of CYP17A1, CYP11A1, and 3β-HSD proteins in mouse Leydig cells, decreasing the expression of StAR protein in mouse Leydig cells, increasing the activity of steroidogenic nuclear receptor promoter, and increasing the secretion of testosterone in mouse Leydig cells.

6. The use according to claim 5, characterized in that The steroidogenic nuclear receptor promoter includes the SF-1 promoter.

7. Application of cinnamaldehyde in the preparation of a functional feed additive for enhancing the expression of 3β-hsd, CYP11A1, and StAR proteins and increasing androgen secretion.

8. The use according to claim 7, characterized in that The dosage of additives in feed is 25-75 mg / kg.

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