【摘要】：骨質(zhì)疏松癥（Osteoporosis，OP）是多種原因引起的一種以骨組織含量減少，骨的微觀結(jié)構(gòu)退化為特征，致使骨的脆性增加、強(qiáng)度降低而導(dǎo)致易于發(fā)生骨折的一種全身性骨骼疾病。骨質(zhì)疏松癥已經(jīng)不僅僅是一種疾病，目前已成為一個(gè)嚴(yán)重的社會(huì)問(wèn)題。它帶來(lái)的最主要癥狀有劇烈的疼痛、多發(fā)骨折以及胸腔縮小引起的呼吸困難。在眾多承受骨質(zhì)疏松癥病痛折磨的患者中，絕經(jīng)后女性因受到衰老和體內(nèi)激素水平下降的雙重影響，成為了該病最大的受害人群，約占總患病人數(shù)70%以上。另外，隨著糖尿病患者日漸增多，特別是中老年人居多，因糖尿病引起的骨質(zhì)疏松癥也成為目前臨床上骨質(zhì)疏松癥形成的重要原因之一，并且由于其具有隱匿性而往往被患者忽視，延誤治療，，其后果不堪設(shè)想。目前防治骨質(zhì)疏松癥的藥物主要是骨吸收抑制劑、骨形成促進(jìn)劑和礦化藥物，這些藥物均屬替代治療，在長(zhǎng)期使用過(guò)程中，這些藥物具有毒副作用大，靶向性低，依從性差等缺點(diǎn)。因此，有必要找尋到一種能夠具有廣泛受眾人群的藥物來(lái)治療骨質(zhì)疏松癥。 中藥的應(yīng)用在我國(guó)已有幾千年的歷史，在人們心中具有極高的認(rèn)同度。中藥在治療骨質(zhì)疏松癥方面具有全身調(diào)理、毒副作用小、取材方便等優(yōu)勢(shì)，越來(lái)越受到人們的關(guān)注。而在眾多中藥中，冬蟲夏草特有的藥用價(jià)值長(zhǎng)久以來(lái)一直受到人們的青睞。因此，如何發(fā)掘出冬蟲夏草的潛質(zhì)，研發(fā)出具有自主知識(shí)產(chǎn)權(quán)的抗骨質(zhì)疏松中藥，成為了我們研究的目的所在。那么，冬蟲夏草是否具有預(yù)防和治療骨質(zhì)疏松癥的能力呢？它發(fā)揮拮抗骨質(zhì)疏松作用的機(jī)制是什么？作為冬蟲夏草重要單體成分的蟲草多肽是否具有抗骨質(zhì)疏松的潛力呢？我們將圍繞著上述疑問(wèn)展開研究。 1.富含鍶的冬蟲夏草對(duì)去卵巢骨質(zhì)疏松大鼠的作用及機(jī)制研究 （1）目的：研究富含鍶的冬蟲夏草對(duì)去卵巢大鼠骨質(zhì)疏松的治療作用和相關(guān)機(jī)制。 （2）方法：將雌性大鼠隨機(jī)分為5組：假手術(shù)組（Sham）、去勢(shì)組（OVX）、冬蟲夏草組（CS）、鍶組（SR）以及富含鍶的蟲草組（CSS）。每組10只。4組通過(guò)手術(shù)摘除卵巢(OVX)，另一組進(jìn)行假手術(shù)（對(duì)照）。第1組（假手術(shù)）和第2組(OVX)經(jīng)口接受10ml生理鹽水治療，第3組、第4組和第5組分別經(jīng)口接受10ml CSS、CS和SR治療，時(shí)間長(zhǎng)達(dá)8周。觀察大鼠尿鈣、血鈣、血磷、骨礦物質(zhì)密度（BMD）、骨礦物質(zhì)含量（BMC）以及子宮、胸腺和體重的變化情況。測(cè)定各組大鼠血清堿性磷酸酶(ALP)、抗酒石酸酸性磷酸酶(TRAP)、血清骨鈣素(OC)、同型胱氨酸（HCY）、I型膠原交聯(lián)型羧基端肽(CTX)、雌二醇和干擾素-γ (IFN-γ)水平。 （3）結(jié)果：CSS和SR均對(duì)去卵巢骨質(zhì)疏松大鼠的骨質(zhì)機(jī)械強(qiáng)度和礦物質(zhì)含量產(chǎn)生了積極的影響。但是，CSS治療組的股骨頸強(qiáng)度高于SR治療組。CSS和SR可顯著降低尿鈣、血鈣以及血磷水平。相反， CS和CSS可顯著增加萎縮子宮的重量和體重，并且還減輕動(dòng)物胸腺質(zhì)量，而SR沒有表現(xiàn)出任何此類作用。同時(shí)CSS可以顯著降低血清ALP和TRAP水平并促進(jìn)骨鈣素的分泌并降低CTX水平。另外，CSS可以明顯降低IFN-γ水平并顯著提高體內(nèi)雌二醇的水平。 （4）結(jié)論：本研究證明了富含鍶的冬蟲夏草在治療人類絕經(jīng)后骨質(zhì)疏松癥中的價(jià)值。CSS可以有效治療因骨強(qiáng)度和骨礦含量下降引起的骨質(zhì)疏松癥，其作用機(jī)制主要是通過(guò)減少骨質(zhì)疏松模型中骨質(zhì)的流失，顯著減少骨吸收，促進(jìn)骨生成并促進(jìn)體內(nèi)雌二醇生成實(shí)現(xiàn)的。同時(shí)也證明了冬蟲夏草同樣具有拮抗骨質(zhì)疏松癥的潛力。 2.冬蟲夏草提取物對(duì)大鼠廢用性骨質(zhì)疏松的預(yù)防作用 （1）目的：驗(yàn)證冬蟲夏草是否具有預(yù)防大鼠廢用性骨質(zhì)疏松癥的作用。 （2）方法：將大鼠隨機(jī)分為6組，其中5組做后肢懸吊（HLS）處理。其中1個(gè)HLS組經(jīng)口給予阿侖膦酸鈉（2.0mg/kg/天），另外3個(gè)HLS組分別經(jīng)口給予不同劑量的冬蟲夏草（100、300、500mg/kg/天），連續(xù)8周，余下的HLS組作為HLS之前和之后不給藥的對(duì)照組。每組分別由10只雄性和雌性組成。對(duì)體重、血清和尿液中的生化指標(biāo)、骨礦物質(zhì)密度（BMD）、骨礦物質(zhì)含量（BMC）、機(jī)械力學(xué)和骨微結(jié)構(gòu)進(jìn)行測(cè)定。 （3）結(jié)果：與其它HLS組相比，使用較高劑量（300和500mg/kg/天）冬蟲夏草或阿侖膦酸鈉的治療組在體重、機(jī)械強(qiáng)度、BMD和BMC等方面產(chǎn)生了積極作用。在HLS大鼠中，冬蟲夏草呈劑量依賴性地使骨轉(zhuǎn)換標(biāo)志物下降，并使骨鈣素水平升高。L-4椎體的顯微CT分析結(jié)果表明，冬蟲夏草高劑量組（500mg/kg）大鼠的骨小梁無(wú)論是數(shù)目、厚度還是骨小梁的間隙都比HLS組得到了明顯的改善。 （4）結(jié)論：本研究表明，經(jīng)過(guò)8周使用較高劑量的冬蟲夏草可以預(yù)防大鼠廢用性骨質(zhì)疏松癥。意味著冬蟲夏草也可以作為預(yù)防人類廢用性骨質(zhì)疏松癥的替代療法。 3.蟲草多肽對(duì)四氧嘧啶誘發(fā)的糖尿病大鼠骨質(zhì)疏松的作用研究 （1）目的：研究蟲草多肽對(duì)四氧嘧啶誘發(fā)的糖尿病大鼠骨質(zhì)疏松癥的保護(hù)作用及可能的機(jī)制。 （2）方法：將大鼠隨機(jī)分為5組，其中4組給予腹腔注射四氧嘧啶構(gòu)建糖尿病模型。3組糖尿病大鼠每天接受腹膜內(nèi)注射用蟲草多肽（Cordymin）（20，50，和100mg/Kg/天），共5周。余下的糖尿病組對(duì)照組。每組分別由10只雄性大鼠組成。對(duì)血糖、糖化血紅蛋白（HbA1c）、血清堿性磷酸酶（ALP）、抗酒石酸酸性磷酸酶（TRAP）、胰島素水平、總體抗氧化劑活性（TAOS）、骨礦物質(zhì)密度（BMD）、骨礦物質(zhì)含量（BMC）以及胰島β細(xì)胞損傷情況進(jìn)行測(cè)定和評(píng)估。 （3）結(jié)果：蟲草多肽可以降低血糖、糖化血紅蛋白（HbA1c）、血液中ALP及TRAP水平，并呈劑量依賴性的升高胰島素水平。此外，糖尿病大鼠經(jīng)過(guò)蟲草多肽的治療可以相對(duì)減少胰腺β細(xì)胞的死亡并降低糖尿病大鼠總體抗氧化劑活性。同時(shí)可以增加糖尿病大鼠的BMC和BMD。 （4）結(jié)論：蟲草多肽對(duì)糖尿病引起的骨質(zhì)疏松癥可能具有一定的保護(hù)作用。 綜上所述：以上研究結(jié)果說(shuō)明：冬蟲夏草能夠促進(jìn)成骨、降低骨質(zhì)流失，并且可以有效的修復(fù)了骨質(zhì)疏松對(duì)機(jī)體機(jī)能的損害。而作為冬蟲夏草中重要的單體成分之一的蟲草多肽可以有效緩解糖尿病癥狀，減輕胰腺β細(xì)胞損傷，有效拮抗體內(nèi)氧化劑的活性，增加體重，促進(jìn)體內(nèi)骨鈣的沉積，甚至具有治療因糖尿病引起的骨質(zhì)疏松癥的潛力。這一重大的發(fā)現(xiàn)為我們找尋到了治療骨質(zhì)疏松癥的新的方向，有望拓展對(duì)冬蟲夏草藥理學(xué)功能和抗骨質(zhì)疏松作用的新認(rèn)識(shí)，為今后研發(fā)具有自主知識(shí)產(chǎn)權(quán)的抗骨質(zhì)疏松藥物奠定堅(jiān)實(shí)的理論基礎(chǔ)。
[Abstract]:Osteoporosis (OP) is a kind of systemic bone disease caused by the decrease of bone tissue and the degeneration of bone microstructure, resulting in the increase of bone brittleness and the decrease of strength, which leads to the fracture of a systemic bone disease. Osteoporosis is not only a disease, but has become a serious society. The main symptoms are severe pain, multiple fractures and dyspnea caused by narrowing of the chest. Among the many patients suffering from osteoporosis, postmenopausal women have become the largest victims of the disease, accounting for about 70% of the total number of diseases due to the double effects of aging and decreased hormone levels in the body. On the other hand, with the increasing number of diabetic patients, especially in the middle and old people, the osteoporosis caused by diabetes is also one of the important reasons for the formation of osteoporosis in clinical. And because of its concealment, it is often ignored by the patients, and the result is unimaginable. The medicine for the prevention and treatment of osteoporosis is at present. Substances are mainly bone resorption inhibitors, bone formation accelerators and mineralized drugs. These drugs are all alternative treatments. During the long term use, these drugs have the disadvantages of large toxic and side effects, low targeting and poor compliance. Therefore, it is necessary to find a drug that can be widely used in the treatment of osteoporosis.
The application of traditional Chinese medicine has a history of thousands of years in our country and has a high degree of recognition in people's heart. Traditional Chinese medicine has the advantages of whole body conditioning, small toxic and side effects and convenient selection of materials in the treatment of osteoporosis, and more and more people pay attention to it. In many Chinese medicine, the special medicinal value of winter insect and summer grass has been subject to people for a long time. Therefore, how to discover the latent quality of the Cordyceps sinensis and develop the anti osteoporosis Chinese medicine with independent intellectual property rights has become the purpose of our research. Then, is the ability of Cordyceps sinensis to prevent and treat osteoporosis? What is the mechanism of antagonizing osteoporosis? As important for Cordyceps sinensis Whether the single component of Cordyceps polypeptide has the potential to combat osteoporosis? We will focus on these questions.
Effect and mechanism of 1. strontium rich Cordyceps sinensis on ovariectomized rats
(1) objective: To study the effect and mechanism of Cordyceps sinensis (SR) on osteoporosis in ovariectomized rats.
(2) method: the female rats were randomly divided into 5 groups: the sham operation group (Sham), the castration group (OVX), the Cordyceps sinensis group (CS), the strontium group (SR) and the strontium rich Cordyceps group (CSS). 10 groups of.4 groups in each group were operated on the ovaries (OVX), the other group was sham operation, and the first groups (sham operation) and the second group (OVX) received 10ml physiological saline treatment and third, Fourth groups and fifth groups were treated with 10ml CSS, CS and SR for up to 8 weeks. The changes of urinary calcium, blood calcium, blood phosphorus, bone mineral density (BMD), bone mineral content (BMC), uterus, thymus and body weight were observed. Serum alkaline phosphatase (ALP), tartaric acid phosphatase (TRAP), and Serum Osteocalcin (OC) were measured in each group. Homocysteine (HCY), type I collagen cross-linked carboxy terminal peptide (CTX), estradiol and interferon gamma (IFN- gamma) level.
(3) results: both CSS and SR had a positive effect on the bone mechanical strength and mineral content in ovariectomized rats. However, the femoral neck strength of the CSS treatment group was higher than that of the SR treatment group,.CSS and SR significantly decreased the urinary calcium, blood calcium and blood phosphorus levels. On the contrary, CS and CSS significantly increased the weight and weight of the atrophied uterus, and also had a significant increase in the weight and weight of the atrophied uterus. The quality of the thymus was reduced, and SR did not show any such effect. At the same time, CSS could significantly reduce the level of serum ALP and TRAP, promote the secretion of osteocalcin and reduce the level of CTX. In addition, CSS can significantly reduce the level of IFN- gamma and significantly increase the level of estradiol in the body.
(4) conclusion: This study proves that the value of strontium rich Cordyceps sinensis in the treatment of postmenopausal osteoporosis can effectively treat osteoporosis caused by the decrease of bone strength and bone mineral content. The main mechanism is to reduce bone loss, reduce bone absorption and promote bone formation by reducing bone loss in the osteoporosis model. It also promotes the production of estradiol in vivo. It also proves that Cordyceps has the potential to combat osteoporosis.
Preventive effect of 2. Cordyceps sinensis extract on disused osteoporosis in rats
(1) objective: to test whether Cordyceps sinensis has the preventive effect of disused osteoporosis in rats.
(2) method: the rats were randomly divided into 6 groups, of which 5 groups were treated with rear limb suspension (HLS), of which 1 groups were given alendronate (2.0mg/kg/ days) through the mouth, and the other 3 HLS groups were given different doses of Cordyceps (100300500mg/kg/ days) for 8 weeks, and the remaining HLS group was used as the control group before and after HLS. Do not consist of 10 males and females. Biochemical markers in body weight, serum and urine, bone mineral density (BMD), bone mineral content (BMC), mechanical mechanics, and bone microstructures were measured.
(3) results: compared with other HLS groups, the treatment group using Cordyceps sinensis or alendronate with higher doses (300 and 500mg/kg/ days) had a positive effect on body weight, mechanical strength, BMD and BMC. In HLS rats, Cordyceps decreased dose-dependent bone conversion markers and increased the level of osteocalcin in the.L-4 vertebral body. The results of micro CT analysis showed that the bone trabecula in the high dose group of Cordyceps sinensis (500mg/kg) rats, whether the number, the thickness or the bone trabecular space, had been significantly improved than that of the HLS group.
(4) conclusion: This study shows that the use of Cordyceps sinensis after 8 weeks of high dose of Cordyceps can prevent disuse osteoporosis in rats, which means that Cordyceps sinensis can also be used as a substitute for the prevention of human waste.
Effect of 3. Cordyceps polypeptide on osteoporosis induced by alloxan in diabetic rats: a study of four
(1) objective: To study the protective effect of Cordyceps polypeptide on osteoporosis induced by alloxan in diabetic rats and its possible mechanism.
(2) method: the rats were randomly divided into 5 groups, of which 4 groups were given intraperitoneal injection of four oxouracil to construct diabetes model.3, the diabetic rats were treated by intraperitoneal injection of Cordymin (20,50, and 100mg/Kg/ days) for 5 weeks. The remaining diabetic control group. Each group was composed of 10 male rats respectively. White (HbA1c), serum alkaline phosphatase (ALP), anti tartaric acid acid phosphatase (TRAP), insulin level, total antioxidant activity (TAOS), bone mineral density (BMD), bone mineral content (BMC), and islet beta cell damage were measured and evaluated.
(3) results: Cordyceps can reduce blood sugar, glycated hemoglobin (HbA1c), ALP and TRAP levels in the blood, and increase the level of insulin in a dose dependent manner. In addition, diabetic rats can reduce the death of pancreatic beta cells and lower the overall antioxidant activity of diabetic rats by the treatment of Cordyceps polypeptide. BMC and BMD. in diabetic rats
(4) conclusion: Cordyceps polypeptide may have some protective effects on osteoporosis caused by diabetes.
In summary, the above results show that Cordyceps sinensis can promote osteogenesis, reduce bone loss, and can effectively repair the damage to the body function of osteoporosis. As one of the important monomers in the Cordyceps sinensis, the Cordyceps can effectively alleviate the symptoms of diabetes, reduce the pancreatic beta cell damage, and effectively antagonize it. The activity of the oxidant in the body increases the body weight, promotes the deposition of bone calcium in the body, and even has the potential for the treatment of osteoporosis caused by diabetes. This major discovery provides us with a new direction for the treatment of osteoporosis and is expected to expand the new understanding of pharmacological function and anti osteoporosis effect of Cordyceps sinensis for the future. The research and development of anti osteoporosis drugs with independent intellectual property rights will lay a solid theoretical foundation.
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R285.5

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