本文選題：六月青多糖 + 六月青皂苷�。� 參考：《廣西醫(yī)科大學(xué)》2013年碩士論文

【摘要】：目的：觀察六月青多糖(LYQP)及其皂苷(LYQS)體外抗乙型肝炎病毒(HBV)及體內(nèi)抗鴨乙型肝炎病毒(DHBV)的作用。 方法：(1)體外抗HBV作用：采用MTT法檢測LYQP及LYQS對轉(zhuǎn)染HBV DNA全基因組的HepG2.2.15細胞的半數(shù)毒性濃度(TC50)及最大無毒濃度(TCo)；采用直接加藥法觀察在TCo條件下不同濃度的藥物對HepG2.2.15細胞的作用,分別在第72h和第144h收集細胞培養(yǎng)上清液,采用ELISA法檢測上清液中HBsAg和HBeAg的滴度,采用熒光定量PCR(FQ-PCR)法檢測上清液中HBV DNA的含量。(2)體內(nèi)抗DHBV作用：用1日齡廣西麻鴨感染DHBV,7d后用普通PCR法篩選出DHBV感染強陽性鴨。隨機分成8組：LYQP高、中、低劑量組；LYQS高、中、低劑量組；模型組和阿昔洛韋(ACV)陽性對照組。每組10只,每組雛鴨均連續(xù)灌胃14d。分別于用藥前(To)、用藥后7d(T7)、14d(T14)及停藥3d(P3)后采血,采用ELISA法檢測血清中DHBsAg和DHBeAg的滴度,用FQ-PCR法檢測血清中DHBV DNA的含量。同時檢測血清中丙氨酸氨基轉(zhuǎn)移酶(ALT)和門冬氨酸氨基轉(zhuǎn)移酶(AST)活性,HE染色觀察鴨肝臟組織的病理學(xué)變化。 結(jié)果：(1)體外抗HBV作用：①LYQP及LYQS對HepG2.2.15細胞的毒性較低,TC50分別為111.77mg/mL;90.46mg/L; TC0分別為11.88mg/mL;1.59mg/L。②無毒濃度下不同濃度的含藥培養(yǎng)液在HepG2.2.15細胞的培養(yǎng)中可有效的抑制HBsAg及HBeAg的分泌和HBV DNA的合成,抑制作用有明顯的時效及量效關(guān)系。③TI都大于2,是高效低毒的抗HBV藥物。(2)體內(nèi)抗DHBV作用：給藥后,與模型組比較,LYQP及LYQS各劑量組鴨血清中DHBV DNA的含量顯著降低(P0.∞或P0.01),停藥3d后,中、低劑量組及ACV組血清中DHBV DNA的含量有回升現(xiàn)象,高劑量組DHBV DNA的回升現(xiàn)象不明顯。與模型組比較,LYQP及LYQS各劑量組能降低鴨血清中AST及ALT的活性,給藥前后血清中DHBsAg及DHBeAg的OD值變化與DNA的含量改變相似；藥物的抗病毒效果與劑量大小及時間有一定關(guān)系。此外,鴨肝臟組織病理學(xué)檢查發(fā)現(xiàn)LYQP及LYQS對DHBV引起的肝損傷有明顯的保護作用。 結(jié)論：LYQP及LYQS體內(nèi)外均有顯著的抗HBV作用,同時能減輕DHBV所致的鴨肝損傷,改善肝功能。
[Abstract]:Aim: to observe the effects of LYQP) and its saponins (LYQS) on anti-hepatitis B virus (HBV) in vitro and anti-duck hepatitis B virus (DHBV) in vivo. Methods in vitro anti-HBV effect: MTT assay was used to detect the median toxic concentration (TC50) of LYQP and LYQS on HepG2.2.15 cells transfected with the whole genome of HBV DNA, and the maximum non-toxic concentration (TCoC) to detect the toxicity of LYQP and LYQS to HepG2.2.15 cells transfected with HBV DNA. The effects of different concentrations of drugs on HepG2.2.15 cells were observed by direct addition method. The supernatants of cell culture were collected at 72h and 144h respectively. The titers of HBsAg and HBeAg in the supernatants were detected by enzyme-linked immunosorbent assay (Elisa). The anti-DHBV activity in the supernatant was detected by fluorescence quantitative PCR- FQ-PCR method. The DHBV positive ducks were screened by normal PCR method after 1 day old Guangxi duck infected with DHBV for 7 days. They were randomly divided into 8 groups: high, middle and low dose groups with high, middle and low doses of LYQS, model group and acyclovir ACV-positive control group. Ten ducklings in each group were fed continuously for 14 days. The serum DHBsAg and DHBeAg titers were detected by Elisa and the contents of DHBV DNA in serum were detected by FQ-PCR. The activities of alanine aminotransferase (alt) and aspartate aminotransferase (AST) in serum were also detected. The pathological changes of duck liver were observed by HE staining. Results the antiHBV effects of 1: 1LYQP and LYQS on HepG2.2.15 cells in vitro were lower. TC50 was 111.77 mg / mL1 / mL 90.46 mg / L, respectively, and TC0 was 11.88 mg / mL1.59mg / L.2 in HepG2.2.15 cells, respectively, which could effectively inhibit the secretion of HBsAg and HBeAg and the synthesis of HBV DNA in HepG2.2.15 cells. The inhibitory effect has obvious time-effect and dose-effect relationship. 3TI is more than 2, which is a high efficiency and low toxicity anti-HBV drug. Compared with the model group, the serum DHBV DNA content of the DHBV DNA in the DHBV DNA of the DHBV group and the LYQS group was significantly lower than that of the model group. After 3 days of withdrawal, the serum DHBV DNA content in the low dose group and the ACV group was increased, but the DHBV DNA level in the high dose group was not obvious. Compared with the model group, LYQP and LYQS could decrease the AST and alt activity in duck serum, and the changes of OD value of DHBsAg and DHBeAg in serum were similar to the change of DNA content before and after administration, and the antiviral effect of the drug was related to the dose and time. In addition, duck liver histopathology showed that LYQP and LYQS had obvious protective effects on DHBV induced liver injury. Conclusion both in vivo and in vitro, WW LYQP and LYQS have significant anti-HBV effects, and can alleviate the damage of duck liver induced by DHBV and improve liver function.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R285.5;R512.62

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