Application of isaria feline in preparation of medicine for treating postmenopausal osteoporosis

Abstract

The invention discloses an application of isaria feline in preparation of a medicine for treating postmenopausal osteoporosis, and an application of isaria feline in preparation of a medicine for treating obesity caused by the postmenopausal osteoporosis. Through influence science, histology and mechanical detection, the bone loss of a postmenopausal osteoporosis rat subjected to gavage treatment of the isaria felina is obviously improved, and meanwhile, the isaria felina is found to be capable of effectively controlling obesity caused by the postmenopausal osteoporosis. Therefore, the isaria feline can be applied to preparation of the medicine for treating the postmenopausal osteoporosis and obesity caused by the postmenopausal osteoporosis.

Description

Technical Field

[0001] The invention belongs to the use of C. cati, and in particular relates to the use of C. cati in preparing a medicine for treating postmenopausal osteoporosis. Background Art

[0002] Postmenopausal osteoporosis is the most common primary osteoporosis. It is a systemic metabolic disease caused by estrogen deficiency, which leads to bone loss and bone microstructure damage, resulting in an increased risk of fractures. It poses a huge threat to the health of women around the world. Clinically, the prevention and treatment of osteoporosis mainly rely on drugs such as bisphosphonates, calcitonin and estrogen. Although they are effective in preventing and treating osteoporosis, their side effects and adverse reactions limit their long-term application.

[0003] At present, the application of traditional Chinese herbal medicine in osteoporosis has attracted more and more attention from researchers and institutions around the world. Isaria felina (IF) is isolated and cultured from Cordyceps sinensis. The strain has been collected and preserved by the General Microbiological Center of the China Microbiological Culture Collection Administration, with the collection number CGMCCNO.0706. Previous studies have confirmed that Isaria felina has immunomodulatory function, but whether it has anti-osteoporosis effect has not been reported in the literature. Summary of the invention

[0004] According to one aspect of the present invention, provided is the use of C. felis spores in preparing a drug for treating postmenopausal osteoporosis.

[0005] According to another aspect of the present invention, provided is the use of C. felis spores in the preparation of a medicament for treating obesity caused by postmenopausal osteoporosis.

[0006] According to another aspect of the present invention, a drug for treating osteoporosis is provided, wherein the active ingredient of the drug comprises mycelium of Coryneporium felis.

[0007] The present invention finds that the bone loss of postmenopausal osteoporosis rats treated with cat spores by intragastric administration is significantly improved through impact, histological and mechanical tests, and also finds that cat spores can effectively control obesity caused by postmenopausal osteoporosis. Therefore, cat spores can be used to prepare drugs for treating postmenopausal osteoporosis and obesity caused by it. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Figure 1 (A) Micro-CT 3D reconstructed images of femoral trabeculae and cortical bone of rats in each group. Figure 1(BG) Trabecular and cortical bone parameters of femurs of rats in each group, including BMD, BV / TV, Tb.N, Tb.sp, CT.Th and CT.V. Data are expressed as mean ± SEM. n = 7, * P < 0.05, ** P < 0.01, *** P < 0.001.

[0009] Figure 2 (A) HE staining observation of distal femoral metaphysis bone tissue of rats in each group. Figure 2 (B) is the trabecular bone area ratio. Figure 2 (C) is the number of osteoblasts. Figure 2 (D) is the number of osteoclasts. Figure 2 (E) TRAP staining of distal femoral metaphysis bone tissue of rats in each group. Data are expressed as mean ± SEM. n = 7, * P < 0.05, ** P < 0.01, *** P < 0.001.

[0010] Figure 3 Comparison of three-point bending test parameters among groups, (A) is the maximum load, (B) is the limit radius. Data are expressed as mean ± SEM. n = 7, * P < 0.05, ** P < 0.01, ns represents no significance.

[0011] Figure 4 (A) Body weight monitoring of rats in each group. Figure 4 (B) Uterine weights of rats in each group after oral administration of C. felis spores. Data are expressed as mean ± SEM. n = 7, ****P < 0.0001. DETAILED DESCRIPTION

[0012] Immune overactivation is one of the main pathogenic factors of postmenopausal osteoporosis caused by estrogen deficiency. At the same time, intestinal flora also plays an important role in bone immunity, which can regulate the balance of Th17 and Treg cells through the immune system, regulate osteoblastic and osteoclast functions, and thus affect bone metabolism. Based on this, this embodiment studies the bone loss of postmenopausal osteoporosis rats treated with cat spore gavage through imaging, histology and mechanical detection, and studies the control of obesity caused by cat spores in postmenopausal osteoporosis.

[0013] The Isaria felina (IF) involved in this embodiment has been collected and preserved by the General Microbiological Center of China Microbiological Culture Collection Administration Committee, and the preservation number is CGMCCNO. 0706. The oral Isaria felina described in this embodiment is the powder after the mycelium of Isaria felina is crushed.

[0014] The preparation method of mycelial powder of Corymborhinus cati is as follows: millet, sucrose, MgSO 4 , KH 2 PO 4 and distilled water to make a solid culture medium; encapsulate the solid culture medium in a culture bottle, seal it with kraft paper, and sterilize it at 120℃-130℃; after cooling to room temperature, inoculate the cat's Coryneformis strain; place the inoculated culture bottle in a well-ventilated environment with a temperature of 20℃-25℃ and a relative humidity of 40%-60% for fermentation and culture. After the mycelium has grown all over the culture bottle, collect the culture and dry it in a drying oven to form mycelium blocks, then use a grinder to make powder, and sieve to obtain the cat's Coryneformis mycelium powder for standby use. The above solid culture medium includes 840 parts of millet, 42 parts of sucrose, and MgSO 4 2.8 parts, KH 2 PO 4 5.5 servings.

[0015] The experimental animals in this embodiment are female Sprague-Dawley rats (SD rats), which were purchased from the Experimental Animal Center of Shanxi Cancer Hospital. 21 12-week-old female Sprague-Dawley rats were raised in an SPF environment, with a 12-hour light cycle, and fed with sterile food and autoclaved water. After feeding under these conditions for 1 week, the SD rats were randomly divided into a control group (CON), a postmenopausal osteoporosis group (OVX), and a cat stick spore treatment group (IF+OVX), with 7 rats in each group, and the OVX group and the IF+OVX group rats were ovariectomized to construct a postmenopausal osteoporosis animal model. After the model was successfully constructed, it was continued to be raised under the above conditions for 12 weeks, during which the IF+OVX group was given 100 mg / kg of cat stick spores by gavage every day, and the remaining groups were given an equal amount of normal saline. After 12 weeks, the rats were made unconscious and killed by inhalation of carbon dioxide, and samples were collected for further analysis.

[0016] 1. Oral administration of C. felis improves bone loss in postmenopausal osteoporotic rats 1.2 Radiographic evaluation Micro-CT scans were performed on the femurs of rats in each group, and the results showed that oral administration of Felis catifolia could improve bone loss in postmenopausal osteoporosis. Micro-CT 3D reconstructions of trabecular and cortical bone in each group are shown in Figure 1 In A, the Micro-CT parameters of trabecular bone and cortical bone are shown in Figure 1 In BG, these include BMD, BV / TV, Tb.N, Tb.sp, CT.Th and CT.V. From the parameter index results, it can be found that oral administration of cat spores can significantly improve the trabecular bone related indicators of OVX rats, but has no obvious effect on the cortical bone.

[0017] 1.2 Histological evaluation In order to study the effect of oral administration of Felis cati on bone morphology in OVX rats, the femur tissue of OVX rats was further examined by histopathological examination, including HE staining and TRAP staining. The results showed that compared with the control group, the femoral trabeculae of OVX rats were broken and sparse ( Figure 2 A), the trabecular area ratio and the number of osteoblasts were significantly decreased ( Figure 2 In addition, there were a large number of osteoclasts on the surface of trabeculae in OVX rats, showing obvious bone resorption. The number of osteoclasts on the surface of trabeculae in IF+OVX group decreased, tending to be similar to that in CON group ( Figure 2 D, E). Oral administration of Felis catifolia can effectively inhibit osteoclasts, promote osteogenesis, and improve bone loss caused by postmenopausal osteoporosis.

[0018] 1.3 Mechanical evaluation The femurs of rats in each group were stress tested using a three-point bending test. Compared with the CON group, the maximum load borne by the femurs of rats in the OVX group was significantly reduced, and the maximum load borne by the femurs of rats in the IF+OVX group improved and tended to the CON group. There was no significant difference in the ultimate deflection between the groups ( Figure 3 A, B).

[0019] 2. Oral administration of cat spores effectively controls obesity caused by postmenopausal osteoporosis During the oral treatment with C. felis spore, the body weight of rats in each group was monitored regularly. Compared with the CON group, the average body weight of rats in the OVX group was significantly higher, while the average body weight of rats in the IF+OVX group was significantly lower than that in the OVX group ( Figure 4 A), indicating that C. felis can effectively control obesity caused by postmenopausal osteoporosis, which may be one of the factors that C. felis improves bone loss in postmenopausal osteoporosis. After the oral C. felis treatment, the uterus of each group of rats was weighed. Compared with the CON group, the uterine weight of the OVX group and the IF+OVX group was significantly reduced ( Figure 4 B), indicating that the OVX rat model was successfully established and oral administration of Felis sporea had no effect on uterine atrophy.

[0020] Studies have shown that there is a causal relationship between obesity and osteoporosis. Increased obesity is harmful to bone microstructure, which may be a risk factor for osteoporosis. Obesity can cause bone marrow mesenchymal stem cells to differentiate into adipocytes, leading to an increase in bone marrow adipose tissue and a decrease in osteoblasts. Excessive bone marrow adipose tissue can change the bone microenvironment and microstructure by replacing osteocytes with adipocytes, thereby leading to a decrease in bone density. At the same time, excessive adipose tissue can release a large number of inflammatory factors, most of which can trigger the RANK pathway, activate osteoclasts and cause bone loss. The present invention found that there were a large number of lipid droplets deposited in the bone marrow cavity of postmenopausal osteoporosis rats in the femur histological detection. The number of lipid droplets in the bone marrow cavity was reduced after treatment with cat club spores. The effective control of cat club spores on obesity also reflects the role of treating osteoporosis from the side.

Claims

1. Application of Felis catifolia in the preparation of drugs for the treatment of postmenopausal osteoporosis.

2. Application of Felis sporea in the preparation of medicine for treating obesity caused by postmenopausal osteoporosis.

3. A drug for treating osteoporosis, characterized in that: Its active ingredients include the mycelium of Corymborhinus felis.

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