本文選題：鹿源 + 結(jié)核菌　； 參考：《吉林農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：結(jié)核病(Tuberculosis,TB)是由結(jié)核分枝桿菌(Mycobacterium tuberculosis)引起的慢性消耗性、人畜共患的重大傳染性疾病�？菇Y(jié)核化學(xué)藥物用于臨床治療結(jié)核病已有數(shù)十年,加之用藥不合理、AIDS的流行等一系列原因,導(dǎo)致了耐藥結(jié)核菌的產(chǎn)生。耐藥結(jié)核病的治療困難,治愈率較低,治療過程中對機體傷害極大。尋找新型治療藥物和治療方案迫在眉睫。中藥被用于治療疾病歷史悠久,其作用細菌機制復(fù)雜,不易使細菌產(chǎn)生耐藥性,且毒副作用較小。所以,本試驗旨在豐富的中藥中尋找對耐藥結(jié)核菌的抑制劑或抗菌增效劑,并對其機理進行初步研究和探討。本實驗對多株結(jié)核菌的耐藥性進行檢測,結(jié)果表明MT-7、MB-1兩株結(jié)核菌為耐多藥結(jié)核菌。因此選擇MT-7、MB-1為本實驗的實驗菌株,以敏感菌株H37Rv作為對照。對17種中藥分別進行水提和醇提,得到17種中藥的水提物和醇提物,以微量稀釋法作用于三株結(jié)核菌,結(jié)果表明,白屈菜、黃芩、蒲公英三種中藥具有較好的體外抗結(jié)核菌活性,其中白屈菜提物效果最好。將三種中藥的提取物與抗結(jié)核藥物聯(lián)合作用于三株結(jié)核菌,結(jié)果表明,白屈菜醇提物與利福平聯(lián)合作用于H37Rv、MB-1、MT-7時,兩者間具有一定的協(xié)同作用。提取白屈菜中總生物堿并純化。測定白屈菜水提物與醇提物中總生物堿含量,通過微量稀釋法比較白屈菜總生物堿與水提物、醇提物的體外抑結(jié)核菌活性。結(jié)果表明,白屈菜總生物堿抑菌活性最好,白屈菜醇提物中生物堿含量比水提物中生物堿含量高,體外抗結(jié)核效果也比水提物效果好。將白屈菜總生物堿與抗結(jié)核藥物聯(lián)合作用于三株結(jié)核菌,結(jié)果表明兩者間具有協(xié)同作用,FIC≤0.5,說明白屈菜總生物堿為利福平的抗結(jié)核菌增效劑。此外,對白屈菜總生物堿對細胞毒性進行了檢測,白屈菜總生物堿的MIC值遠遠低于白屈菜總堿對細胞有毒性作用的濃度值。用白屈菜總生物堿、白屈菜總生物堿和利福平分別作用于三株結(jié)核菌,提取結(jié)核菌的RNA,以熒光定量PCR方法檢測結(jié)核菌PhoP、PhoR基因的表達量。檢測結(jié)果表明,在高濃度白屈菜總生物堿作用下,兩株耐藥結(jié)核菌的PhoP、PhoR基因表達量均有下降,說明白屈菜總生物堿可以降低耐藥結(jié)核菌的耐藥性。在白屈菜總生物堿和利福平聯(lián)合作用下的兩株耐藥結(jié)核菌的PhoP、PhoR基因表達量也均有有明顯下降,說明白屈菜生物堿和利福平間存在協(xié)同作用,也證實了白屈菜生物堿是利福平的抗結(jié)核菌增效劑,其抗結(jié)核菌增效機制有待進一步研究。
[Abstract]:Tuberculosis (TB) is a serious infectious disease caused by Mycobacterium tuberculosism. Antituberculous chemicals have been used in clinical treatment of tuberculosis for decades, and a series of reasons, such as irrational use of drugs and the prevalence of AIDS, have led to the emergence of drug-resistant TB bacteria. Drug-resistant tuberculosis treatment is difficult, the cure rate is low, the treatment process is very harmful to the body. It is urgent to look for new therapeutic drugs and therapeutic schemes. Chinese medicine has been used to treat diseases for a long time, its mechanism is complex, it is not easy to make bacteria drug resistance, and the toxicity and side effects are relatively small. Therefore, the purpose of this study was to search for inhibitors or synergists against drug-resistant tuberculous bacilli in Chinese medicine, and to explore its mechanism. The results showed that two strains of MT-7 MB-1 were multidrug resistant. Therefore, MT-7 MB-1 was selected as the experimental strain and the sensitive strain H37Rv as control. The water and alcohol extracts of 17 kinds of traditional Chinese medicine were extracted by water and alcohol, respectively, and three strains of tuberculous bacilli were treated with microdilution method. The results showed that Scutellaria baicalensis. Dandelion has good anti-tuberculous activity in vitro. Three strains of tuberculous bacteria were treated with the extracts of three kinds of traditional Chinese medicine and antituberculous drugs. The results showed that there was a certain synergism between the alcohol extract and rifampicin in H37Rvv1 MB-1 MT-7. Total alkaloids were extracted from Dioscorea chinensis and purified. The contents of total alkaloids in water extract and alcohol extract were determined and the antibacterial activity of alcohol extract against tuberculous bacteria in vitro was compared by microdilution method. The results showed that the total alkaloids of Dioscorea albicans had the best antibacterial activity. The alkaloid content in the alcohol extract was higher than that in the water extract, and the antituberculous effect in vitro was better than that in the water extract. Three strains of tuberculous bacteria were treated with total alkaloids and antituberculous drugs. The results showed that there was a synergistic effect of FIC 鈮，

本文編號：1912113