雄黄及含雄黄复方对应激蛋白HSP70和HO-1的影响

Effects of realgar and prescriptions containing realgar on stress response proteins HSP70 and HO-1

目的:研究雄黄和含雄黄复方对病理状态下应激蛋白(热休克蛋白70、血红素氧合酶-1)的影响,探讨雄黄的"解毒"作用机理。方法:制备大鼠感染性脑损伤模型,雄黄(75mg/kg、150mg/kg、300mg/kg)组、安宫牛黄散(1.5g/kg)组灌胃后3h取血和脑组织;另制备大鼠发热模型,灌胃雄黄(90mg/kg)、牛黄解毒片(1.404g/kg),分别于药后1h、2h、4h组取血。用ELISA法测定脑组织、血清中的热休克蛋白70(HSP70)含量,用双波长分光光度法测定脑组织、血清中的血红素氧合酶-1(HO-1)活力。结果:高剂量雄黄和安宫牛黄散使感染性脑损伤大鼠脑组织中的HSP70含量比模型组显著增加(17.9%、30.5%)。雄黄给药后1h和2h使发热大鼠血清HSP70水平比模型组显著上升(36.1%和18.0%),牛黄解毒片给药后1h、2h和4h使发热大鼠血清HSP70水平显著上升(54.6%、19.8%和21.2%)。雄黄对脑损伤大鼠脑组织和血清中的HO-1活力影响不显著;安宫牛黄散使脑损伤大鼠脑组织和血清中的HO-1活力增加(24.3%和12.1%)。雄黄在药后1h、2h和4h分别使发热大鼠血清HO-1活力比模型组增加25.4%、46.6%和41.0%;牛黄解毒片在药后1h、2h和4h使发热大鼠血清HO-1活力比模型组增加1.07倍、1.53倍和94.8%。结论:一定剂量的雄黄和含雄黄复方可诱导、激活病理状态下大鼠体内的应激蛋白HSP70、HO-1,两种含雄黄复方提升应激蛋白水平的幅度大于雄黄单独给药。雄黄提升病理状态下机体的应激水平可能是其发挥"解毒"作用的途径之一。

Objective： To explore the pharmacological mechanism of Realgar by way of studying the effects of Realgar and prescriptions containing Realgar on stress response proteins heat shock protein （HSP70） and heme oxygenase-1 （ HO-1 ） under pathologic status （inflammatory cerebral injury model and fever model）. Methods： In experiment 1, SD rats were randomly divided into six groups, and 6 rats in each group ： untreated normal group, inflammatory cerebral injury model group, model rats administrated by Realgar of low dosage（75mg/kg）, middle dosage（ 150mg/kg）, high dosage（300mg/kg） and by Angong Niuhuang powder（ ANP, 1.5g/kg）. Blood samples and brain tissues were collected at 3 hrs after administration. In experiment 2, SD rats were randomly divided into four groups, and 15 rats in each group ： untreated normal group, fever model group, model rats administrated by Realgar（90mg/kg）and model rats administrated by Niuhuang Jiedu tablet（ NJT, 1. 404g/kg）. Each group was divided into three subgroups（5 rats/subgroup）. Blood samples of the rats in subgroups were collected at 1hr, 2hrs and 4hrs after administration respectively. ELISA method was used to determine HSP70 levels in serum and brain tissues, and dual wavelength spectrophotometry was used to determine HO-1 activities in serum and brain tissues. Result： High dose Realgar and ANP increased HSP70 levels in brain tissues by 17.9% （ P 〈 0.01 ） and 30.5% （ P 〈 0.01 ） apparently comparing to cerebral injury model group. Realgar obviously increased HSP70 levels in serum by 36.1% （ P 〈 0.01 ） and 18.0% （ P 〈 0.01 ） at 1 h and 2h after administration comparing to fever model group. NJT obviously increased HSP70 levels in serum by 54.6% （ P 〈 0.01 ）, 19.8% （ P 〈 0.01 ） and 21.2% （ P 〈 0.01 ） 1h, 2h and 4h after administration comparing to fever model group. Realgar had no obvious effect on activities of HO-1 in brain and serum comparing to cerebral injury model group. ANP enhanced HO-1 activities in brain and serum by 24.3 % （ P 〈 0.01 ） and 12.1% （ P 〈 0.05） comparing to cerebral injury model rats. Realgar enhanced HO-1 activity in serum by 25.4% （ P 〈 0.01 ）, 46.6% （ P 〈 0.01 ） and 41.0% （ P 〈 0. 01 ） 1h, 2h and 4h after administration comparing to fever model rats. NJT enhanced HO-1 activity in serum by 1.07 times（ P 〈0.01 ）, 1.53 times（ P 〈0.01 ）and 94.8% （ P 〈0.01 ）at 1h, 2h and 4h after administration comparing to fever model group. Conclusion： Realgar of certain dosage and prescriptions containing Realgar can induce synthesis of HSP70 and enhance activity of HO-1 in rats under pathological status. Two prescriptions containing Realgar（ ANP, NJT） increase HSP70 and HO-1 to higher level or activity than those increased byRealgar. The experimental test-data provide some evidence which supports the author＇s assumption that Realgar as contained in certain prescriptions, at certain specific levels, assists in the removal of internal toxins by inducing stress protein to improve the positive stress level in the body under pathologic status.