Clinical Observation Of Qingda Granules In The Treatment Of Hypertension And Study Of The Mechanism Of Regulating NLRP3-mediated Cell Pyroptosi

BackgroundHypertension can lead to fatal cardio-cerebrovascular diseases such as myocardial infarction,heart failure and stroke,which is an important global public health problem in the world.Although many antihypertensive drugs have made great progress,the hypertension of most patients has not been fully controlled.Therefore,it is still a major clinical demand to develop a new treatment scheme for the prevention and treatment of hypertension.Qingda Granule is further refined on the basis of Qingxuan Jiangya decoction,which has been used by Academician Chen Keji in the treatment of hypertension for more than 60 years.In the past,a single-center randomized controlled clinical study confirmed that Qingda Granule could control blood pressure(BP)and relieve the symptoms of grade 1 hypertension patients with hyperactivity of liver fire,but this clinical study did not use blind method.Previous experimental studies have found that Qingda Granule has the effects of relaxing blood vessels,reducing blood pressure,reducing inflammation and improving cardiovascular remodeling.ROS/NLRP3-mediated pyroptosis is closely related to hypertension and target organ damage,but it is not clear whether Qingda Granule can interfere with hypertension and target organ damage by regulating ROS/NLRP3-mediated pyroptosis.Objective1.Summarize the academic thoughts and clinical medication experience of traditional Chinese medicine(TCM)great master Academician Chen Keji on the prevention and treatment of hypertension through medical record data mining.2.To evaluate the efficacy and safety of Qingda Granule in the treatment of grade 1 essential hypertension by a randomized double-blind parallel controlled clinical trial.3.Based on ROS/NLRP3-mediated pyroptosis pathway to analyze the mechanism of Qingda Granule in inhibiting cardiovascular remodeling in spontaneously hypertensive rats(SHR).Method1.Using the Ancient and Modern Medical Case Cloud Platform(WEB terminal)and the Traditional Chinese Medicine Inheritance Computing Platform(V3.0)to conduct statistical analysis on the clinical medical records of TCM great master Academician Chen Keji in the treatment of hypertension.First,the frequency statistics method was used to analyze the distribution of age,sex,symptoms,tongue,pulse and TCM syndromes of hypertensive patients treated by Academician Chen Keji.In addition,frequency statistics,correlation analysis and cluster analysis were used to further summarize the combination rule of TCM such as distribution of high-frequency TCM and its efficacy,the four characters,five tastes and meridian distributions,high-frequency paired medicines and angle medicines.At the same time,the core prescription of Academician Chen Keji in the treatment of hypertension was extracted and the diagnosis and treatment ideas of Academician Chen Keji in the treatment of hypertension were summarized.2.This clinical study was a single center part of Xiyuan Hospital,China Academy of Chinese Medical Sciences from a multi-center,prospective randomized,double-blind,doublesimulation,parallel controlled clinical trial involving 552 patients from 13 hospitals in China.The patients with liver-yang hyperactivity syndrome of grade 1 essential hypertension were randomly divided into the treatment group and the control group according to 1:1.The subjects in the treatment group took Qingda Granule+Valsartan Capsule simulant,Qingda Granule 1 bag/time,twice a day.Valsartan Capsule simulant,1 capsule/day,control group took Qingda Granule simulant+Valsartan Capsule,Qingda Granule simulant 1 bag/time,twice a day,Valsartan Capsule,1 capsule/day,and the course of treatment was 4 weeks.Efficacy indicators:changes in office blood pressure,ambulatory blood pressure,hypertension-related symptoms,total score of TCM syndromes,Hamilton anxiety score,renin-angiolensin-aldosterone system(RASS)and plasma connective tissue growth factor(CTGF)after 4 weeks treatment.Safety indicators:blood routine,liver function,renal function,urine routine,ECG and adverse events.When the continuous variable conforms to the normal distribution,it is expressed by mean and standard deviation,if it is non-normal distribution,it is expressed by median and quartile spacing,and the classified variables are expressed in the form of quantity and percentage.P<0.05 considered the difference to be statistically significant.3.WKY rats were taken as normal control group(gavage of normal saline).Using SPSS Statistics 20.0,spontaneously hypertensive rat(SHR)were randomly divided into model group,Qingda Granule low-dose group(0.45 g/kg/d gavage),middle-dose group(0.9 g/kg/d gavage),high-dose group(1.8 g/kg/d gavage),NLRP3 inhibitor group(MCC950 10 mg/kg,3 times/week,intraperitoneal injection),NLRP3 activator group(Nigericin 1 mg/kg,3 times/week,intraperitoneal injection+Qingda Granules 0.9 g/kg/d gavage)and positive drug group(Valsartan Capsules 7.2 mg/kg/d gavage))for 8 consecutive weeks.The systolic blood pressure(SBP),diastolic blood pressure(DBP)and mean arterial blood pressure(MBP)of rats were measured every two weeks;Echocardiography were used to detect ventricular structurerelated indicators;HE staining and Masson staining were used to observe the pathomorphological effects of drugs on rat heart and vascular tissue.Serum levels of interleukin-1β(IL-1β),interleukin-18(IL-18),transforming growth factor β1(TGF-β1)and CTGF were detected by ELISA;Immunohistochemical staining was used to detect the expression of type I collagen in rat heart and vascular tissue;DHE staining was used to detect the expression of ROS in heart and vascular tissue.Western Blot were used to detect the expression of pyroptosis-related proteins NLRP3,Cle-caspase-1 and GSDMD in cardiac and vascular tissues.Results1.A total of 90 hypertensive patients treated by TCM great master Academician Chen Keji were included in the medical case study.There were 154 medical records,involving 75 symptoms,9 types of tongue texture,7 types of tongue coating,9 types of pulse,6 types of TCM syndromes and 129 kinds of TCM.The common symptoms and signs are dizziness,insomnia,irritability,headache,constipation,dark tongue,red tongue,pulse strings and so on.The distribution of TCM syndromes is mainly the hyperactivity of liver yang.The nature of commonly used TCM is mainly cold,and the flavor is mainly sweet,bitter and pungent.The meridian of traditional Chinese medicine is mainly liver meridian.The most commonly used paired medicine is "Gastrodia elata-Uncaria".The high-frequency TCM efficacy is mainly based on the method of "clearing heat and calming the liver",and construct the core prescription"Gastrodia elata-Uncaria-Mulberry leaves-Chrysanthemum-Kuding tea-EucommiaAchyranthes bidentata-Prunella vulgaris".2.The results of randomized controlled trial(RCT)showed that there was no statistically significant difference in the baseline indicators between the treatment group and the control group(P>0.05),and they were comparable.,In terms of blood pressure,the treatment group can improve the office blood pressure and reduce the daytime DBP(P<0.05).The 24-hour SBP,24-hour DBP,24-hour MBP,24-hour pulse pressure,daytime SBP,nighttime SBP and nighttime DBP in the treatment group all showed a downward trend after treatment.In terms of the curative effect of TCM syndrome,Hamilton anxiety score and hypertension related symptoms,the treatment group could significantly improve the TCM syndrome score of liveryang hyperactivity and Hamilton anxiety score(P<0.05).It can also significantly improve the symptoms related to vertigo,headache,irritability,waist and knee soreness,tinnitus,palpitations,anxiety,tension and depression(P<0.05).The treatment group was superior to the control group in improving TCM syndrome score of liver-yang hyperactivity,Hamilton anxiety score and symptoms of vertigo,headache,tinnitus,palpitation,anxiety,tension and depression.In terms of RASS,the treatment group could significantly reduce the level of angiotensin Ⅱ(P<0.05).In terms of the index of fibrosis,the treatment group could significantly reduce CTGF(P<0.05).In terms of safety indicators,there was no significant difference in blood routine,liver function,and renal function in the treatment group before and after treatment(P>0.05).The Kappa test of urine routine and electrocardiogram in the treatment group showed homogeneity,and no adverse events occurred.3.The results of the experimental study showed that compared with the normal control group,the SBP,DBP and MBP levels in the model group were significantly increased(P<0.05).Compared with the model group,low,middle and high doses of Qingda Granule,Positive drug group,NLRP3 inhibitor group,NLRP3 activator group could significantly reduce the SBP,DBP and MBP of SHR.The DBP of NLRP3 activator group was significantly higher than that of Qingda Granule Middle Dose(QDG-M)group(P<0.05).Compared with the model group,the levels of TGF-β1 and CTGF in Qingda granule low,middle and high dose groups,positive drug group,NLRP3 inhibitor group and NLRP3 activator group were significantly decreased(P<0.05).The levels of TGF-β1 and CTGF in NLRP3 activator group were significantly higher than those in QDG-M group(P<0.05).The results of echocardiography showed that the thickness of the left ventricular anterior wall and the left ventricular posterior wall thickness in the high-dose Qingda Granules group were significantly lower than those in the model group(P<0.05).The interventricular septum thickness and left ventricular anterior wall thickness in the positive drug group were significantly lower than those in the model group(P<0.05).Masson results showed that the collagen volume fraction(CVF)of myocardium and blood vessels in Qingda granule low,middle and high dose groups,positive drug group and NLRP3 inhibitor group were significantly lower than those in model group(P<0.05).The CVF of myocardial tissue in the NLRP3 activator group was significantly higher than that in the QDG-M group(P<0.05).The results of immunohistochemistry showed that the levels of type I collagen in heart and blood vessels in Qingda granule middle and high dose groups,positive drug group and NLRP3 inhibitor group were significantly lower than those in model group(P<0.05).The level of type I collagen in NLRP3 activator group was significantly higher than that in QDG-M group(P<0.05).The results of DHE staining showed that the levels of ROS in the heart and vascular tissue of Qingda Granule in the middle and high dose groups,the positive drug group,and the NLRP3 activator group were significantly lower than those in the model group(P<0.05).Western Blot results showed that the levels of NLRP3,Cle-caspase-1 and GSDMD in the heart and vascular tissue of Qingda Granule low,medium and high dose groups,positive drug group and NLRP3 inhibitor group were significantly lower than those in the model group(P<0.05).The Cle-caspase-1 of vascular tissue in the NLRP3 activator group was significantly higher than that in the QDG-M group(P<0.05),but there was no significant change compared with the model group(P>0.05).Compared with the model group,the levels of serum IL-1β and IL-18 in Qingda granule low,middle and high dose groups,positive drug group,NLRP3 inhibitor group and NLRP3 activator group decreased significantly(P<0.05).The level of IL-18 in NLRP3 activator group was significantly higher than that in QDG-M group.ConclusionAcademician Chen Keji,a great master of TCM,advocated the treatment of hypertension from the liver and created Qingda Granule composed of Gastrodia elata,Uncaria,Scutellaria baicalensis and Lotus Plumule on the basis of the method of clearing heat and calming the liver.Clinical studies have shown that Qingda Granule can not only lower BP,but also improve hypertension symptoms such as vertigo,headache,irritability,waist and knee soreness,tinnitus,palpitation,anxiety,tension,depression and inhibit RASS system and fibrosis level.The experimental study further confirmed that Qingda Granule can reduce BP and improve cardiovascular remodeling,and the mechanism is related to the inhibition of ROS/NLRP3induced pyroptosis.