本文選題：肺癆康　切入點(diǎn)：耐異煙肼肺結(jié)核　出處：《成都中醫(yī)藥大學(xué)》2016年碩士論文

【摘要】：目的：研究中藥復(fù)方肺癆康治療耐異煙肼肺結(jié)核小鼠的分子機(jī)制,從分子生物學(xué)角度,探討其治療作用與inhA基因突變之間的關(guān)系,為中醫(yī)藥治療耐藥肺結(jié)核提供一些研究思路。方法：本研究先取80只昆明種小鼠,尾靜脈注射法建立耐異煙肼肺結(jié)核動(dòng)物模型,隨機(jī)分為模型組、異煙肼組、肺癆康組及異煙肼聯(lián)合肺癆康組,分別給予生理鹽水、異煙肼、肺癆康、異煙肼聯(lián)合肺癆康灌胃治療56天；選取20只同等小鼠,尾靜脈注射H37RV標(biāo)準(zhǔn)結(jié)核分枝桿菌菌懸液,建立H37RV標(biāo)準(zhǔn)菌小鼠模型,以生理鹽水灌胃56天；選取20只同等小鼠,尾靜脈注射生理鹽水作為空白組,以生理鹽水灌胃56天；比較各組小鼠一般體征及治療前后體重變化,56天后取小鼠肺臟組織進(jìn)行細(xì)菌培養(yǎng)：肉眼及光鏡下觀察各組小鼠肺臟組織病理變化；利用全基因組重測(cè)序技術(shù)對(duì)治療后小鼠肺組織中結(jié)核桿菌基因組DNA進(jìn)行重測(cè)序,以標(biāo)準(zhǔn)菌株(H37RV)為參考序列,篩選對(duì)比分析；采用SPSS21.0統(tǒng)計(jì)軟件對(duì)實(shí)驗(yàn)中的計(jì)量、計(jì)數(shù)資料進(jìn)行統(tǒng)計(jì)學(xué)分析,采用bwa(version 0.7.8)、samtools、Picard等軟件處理全基因測(cè)序后的數(shù)據(jù)并進(jìn)行堿基與基因組表達(dá)結(jié)果的比對(duì)。結(jié)果：通過對(duì)比治療前后的的小鼠體重變化,各組小鼠治療前體重組間差異無統(tǒng)計(jì)學(xué)意義(P0.05),治療后各自小鼠體重組間差異有統(tǒng)計(jì)學(xué)意義(P0.05),標(biāo)準(zhǔn)菌株組與模型組比較,差異無統(tǒng)計(jì)學(xué)意義(P0.05),異煙肼組、肺癆康組、聯(lián)合用藥組與模型組比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05)；肉眼觀察小鼠肺部形態(tài)發(fā)現(xiàn)標(biāo)準(zhǔn)菌株組和模型組肺部形態(tài)有明顯改變,每只小鼠肺組織均有大小不等結(jié)節(jié),異煙肼組、肺癆康組、聯(lián)合用藥組僅僅有部分小鼠肺臟現(xiàn)有結(jié)節(jié),比較各組小鼠結(jié)節(jié)數(shù)差異性,標(biāo)準(zhǔn)菌株與模型組差異無統(tǒng)計(jì)學(xué)意義(P0.05)；異煙肼組與模型組比較差異有統(tǒng)計(jì)學(xué)意義(P0.05)；肺癆康組與模型組比較差異有統(tǒng)計(jì)學(xué)意義(P0.05)；聯(lián)合用藥組與模型組比較差異有統(tǒng)計(jì)學(xué)意義(P0.05),肺癆康、模型組、聯(lián)合用藥組之間差異性無統(tǒng)計(jì)學(xué)意義(P0.05)；光鏡下觀察治療組肺臟炎癥反應(yīng)較模型組明顯減輕；研究發(fā)現(xiàn)與結(jié)核桿菌耐藥基因inhA基因突變相關(guān)的可能的突變堿基位點(diǎn)有三個(gè),分別位于基因組的第1673425、1676290、1678706堿基位置,對(duì)應(yīng)堿基C、C、A突變?yōu)門、A、C。該位置堿基突變分別位于編碼序列上游、下游、下游,基因組編碼序列的改變分別在777、1279、3695。對(duì)比各組堿基位點(diǎn),標(biāo)準(zhǔn)菌株這三個(gè)位置堿基與參考?jí)A基一致,模型組、肺癆康組、異煙肼組、聯(lián)合組耐異煙肼結(jié)核桿菌與inhA對(duì)應(yīng)堿基均有突變,且突變位置、基因組編碼序列、基因一致。結(jié)論：1.肺癆康治療耐異煙肼肺結(jié)核模型小鼠,能顯著改善小鼠的一般體征及組織病變,減輕肺部炎癥反應(yīng),為臨床應(yīng)用治療耐藥肺結(jié)核提供更多依據(jù)；2.通過全基因測(cè)序發(fā)現(xiàn)一些新的與inhA基因突變相關(guān)位點(diǎn),分別位于編碼序列上游、下游、下游,基因組編碼序列的改變分別在777、1279、3695；3.初步研究發(fā)現(xiàn)肺癆康抗耐異煙肼肺結(jié)核的分子機(jī)制與誘發(fā)inhA基因回復(fù)突變關(guān)系不明顯,這為中藥復(fù)方抗結(jié)核的進(jìn)一步研究提供參考。
[Abstract]:Objective : To study the molecular mechanism of traditional Chinese medicine compound pulmonary tuberculosis in the treatment of resistant pulmonary tuberculosis in mice , and to explore the relationship between the therapeutic action and inhA gene mutation , and to provide some research thinking for the treatment of drug - resistant pulmonary tuberculosis in Chinese medicine .
Twenty - one equivalent mice were selected and H37RV standard mycobacterium suspension was injected intravenously to establish a model of H37RV standard strain mice .
Twenty - five mice were treated with normal saline ( normal saline ) as blank group , and saline was used for 56 days .
After 56 days , the lung tissues of mice were cultured for bacterial culture : the pathological changes of lung tissues were observed with naked eyes and under light microscope .
The genomic DNA of mycobacterium tuberculosis in lung tissues of mice after treatment was re - sequenced by full - genome re - sequencing technology , and the reference sequence of standard strain ( H37RV ) was used as reference sequence , and the comparative analysis was screened ;
The results showed that the difference between the two groups was not statistically significant ( P0.05 ) , and the difference was significant ( P0.05 ) .
The lung morphology of mice was observed to be significantly different from the standard strain group and the model group , and the lung tissues of each mouse had a large number of nodules , and the difference between the number of nodules and the standard strain and the model group was not statistically significant ( P0.05 ) .
Compared with the model group , the difference was significant ( P0.05 ) .
There was significant difference between the tuberculosis group and the model group ( P0.05 ) .
There was no significant difference between the combination therapy group and the model group ( P0.05 ) , but there was no significant difference between the two groups ( P0.05 ) .
Under light microscope , the inflammatory response of lung in the treatment group was significantly reduced compared with the model group .
The results showed that 1 . tuberculosis model mice could significantly improve the general physical signs and tissue pathological changes of mice , reduce the inflammatory response of the lung , and provide more basis for the clinical application of drug - resistant pulmonary tuberculosis .
2 . Some new genes related to inhA gene mutation were identified by full - gene sequencing , which were located in the upstream , downstream and downstream of the coding sequence , respectively , and the changes of genomic coding sequence were 777 , 1279 , 3695 , respectively .
3 . It was found that the relationship between the molecular mechanism of pulmonary tuberculosis and the reversion of inhA gene was not obvious , which provided a reference for the further study of the compound anti - tuberculosis of Chinese medicine .

【學(xué)位授予單位】：成都中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R285.5

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 陳曦,馬s，

本文編號(hào)：1668738