黄连生物碱对TGEV诱导的ST细胞炎症反应的调节作用

The inflammatory response regulating role of alkaloids of Coptis chinensis in swine testis cells induced by TGEV

分别用质量浓度为100、50、25μg/m L的黄连生物碱(ACC)处理猪传染性胃肠炎病毒(TGEV)感染的猪睾丸细胞(ST),采用噻唑蓝(MTT)法检测细胞的活力;采用硝酸还原酶法检测NO含量;采用比色法检测总一氧化氮合成酶(TNOS)和诱导型一氧化氮合成酶(i NOS)活性;采用荧光定量PCR检测TNF-α、IL-1β、IL-6、IL-10等炎症基因的表达。结果表明:100、50μg/mL的ACC可极显著提高TGEV感染的ST细胞的存活率,25μg/mL的ACC可显著提高TGEV感染的ST细胞存活率;TGEV感染后ST细胞NO含量急剧升高,100、50μg/mL的ACC可极显著降低ST细胞TNOS和i NOS活性,25μg/mL ACC可显著降低TNOS和i NOS活性;ACC可极显著下调ST细胞TNF-α、IL-1β、IL-6、IL-10m RNA的表达水平。表明黄连生物碱可以降低TGEV诱导的ST细胞的炎症反应,缓解病毒对细胞的损伤。

The swine testicular(ST)cells infected with transmissible gastroenteritis virus(TGEV)were treated with alkaloids of Coptis chinensis(ACC)with mass concentrations of 100,50 and 25μg/mL respectively.MTT assay was used to detect the cell viability,the griess method and colorimetric method were used to detect the release of NO,TNOS and iNOS,and Real-time PCR method was used to detect expression changes of inflammatory genes such as TNF-α,IL-1β,IL-6 and IL-10.The results indicated that ACC at the mass concentration of 100 and 50μg/mL could extremely significantly increase the survival rate of ST cells,and 25μg/mL ACC could significantly increase the survival rate of ST cells.The content of NO increased sharply after TGEV infection,and the ACC at the mass concentration of 100 and 50μg/mL could extremely significantly decrease the activities of TNOS and iNOS,and 25μg/mL ACC could significantly decrease the activities of enzymes TNOS and iNOS.Meanwhile,ACC could extremely significantly down-regulate the expression levels of TNF-α,IL-1β,IL-6 and IL-10 mRNA.We can conclude that the alkaloids of Coptis chinensis could reduce the inflammatory response induced by TGEV and alleviate the damage of virus to cells.