Mechanism Of Maxing Xiongting Decoction On Hypoxia Pulmonary Hypertension Model Rats

ObjectiveTo observe the effect of Maxing Xiongting Decoction on hypoxic pulmonary hypertension in rats model of hypoxic pulmonary hypertension,and also observe the effect of the decoction on Rhodes,HMGB1,NF-κB and VEGF.To explore the mechanism of Maxing Xiongting Decoction in treating hypoxic pulmonary hypertension model rats.Method1.Experimental animals:Sixty Sprague Dawley(SPF)SD rats(body mass250-300 g),male,after adaptive feeding were randomly divided into five groups:the blank control group,the positive control group(fasudil),the model control group(HPH),the high-dose group of Maxing Xiongting Decoction,and the low-dose group of Maxing Xiongting Decoction,with 12 rats in each group.2.Model establishment methods:a rat model of hypoxic pulmonary hypertension(phlegm stasis obstruction lung syndrome of HPH)was established by a combination of swimming,smoky smoking and monocrotaline intraperitoneal injection,and the model cycle was 15 days.For the first 2 weeks,the model control rats were placed in a dedicated constant temperature water cylinder(43±2)℃during forced swimming,30 min per day.After swimming,rats were placed in a 1 m~3animal smoke box for 1 h smoked by a Red Mei card(20 cigarettes)and smoked 1 time per day.Swimming and smoky modeling were performed for 6 days with rest on day 7 for 2 weeks.On the15th day,a rat model of pulmonary hypoxic pulmonary hypertension with phlegm stasis resistance was established by intraperitoneal injection(60 mg/kg)of 1%monocrotaline.3.Method of administration:pharmacologic intervention was started on day 2after monocrotaline intraperitoneal injection,and continuous intervention was performed for 2 weeks.A combined administration mode of gavage and intraperitoneal injection was used in each group.Among them,the blank control group and model control group were treated with 10 ml/kg/d of intragastric and intraperitoneal saline.In the positive control group,fasudil 10 mg/kg/d was administered intragastrically and intraperitoneally.In the high-dose and low-dose groups of Maxing Xiongting Decoction,intragastric administration at 20 g/kg/d,5g/kg/d and intraperitoneal injection of normal saline at 10 ml/kg/d,respectively.4.Specimen collection and detection:the specimens were collected after 14 days of drug intervention.Rats were anesthetized after being weighed,blood was collected from the femoral artery in anticoagulant free blood collection tubes,and the serum was collected by centrifugation after standing for 4 h at room temperature,and Rhodes concentrations in the serum were measured by ELISA.The rats were sacrificed after blood collection,the lungs were removed by cutting the thoracic cavity,weighed,and then the lung tissues were frozen and stored in liquid nitrogen.After extraction of total RNA,the expression of HMGB1,NF-κB and VEGF were detected by RT-PCR.Result:ELISA assay results suggested that Rhodes was significantly higher in the model control group compared with the blank control group,with significant differences(P<0.05).Compared with the model group,the contents of Rhodes in serum of rats in the fasudil group and the Maxing Xiongting Decoction high-dose group were reduced,with significant differences(P<0.05).The results of RT-PCR assay showed that mRNA expression of HMGB1,NF-κB,VEGFof rat lung tissue in the model control group increased significantly(P<0.05),compared with the blank control group.Compared with the model control group,the mRNA expression of HMGB1,NF-κB,VEGF in lung tissue of rats in the Maxing Xiongting Decoction high-dose group was decreased with significant differences(P<0.05).The Maxing Xiongting Decoction low-dose group did not show significant difference and was not statistically significant.While the fasudil group was decreased significantly(P<0.01).ConclusionsMaxing xiongting decoction can improve the general situation of HPH rats with phlegm and blood stasis obstructing lung syndrome,relieve shortness of breath,wheezing and other symptoms.It can reduce the infiltration of inflammatory cells in lung tissue,and significantly reduce the mRNA expression of HMGB1,NF-κB and VEGF,effectively inhibit inflammation.Mechanism of maxing xiongting Decoction in treating HPH rats with phlegm and blood stasis obstructing lung syndrome is related to inhibite HMGB1,NF-κB,VEGF and Rhodes.