Study On The Effect And Mechanism Of Relieving Depression And Resolving Phlegm On Oxidative Stress And Neuroinflammation In Refractory Epileps

Background:Epilepsy is a common chronic central nervous system disorder that causes severe physical and psychological damage and a heavy burden on individuals and their families due to recurrent seizures.About 1/3 patients with epilepsy are resistant to traditional anti-epileptic drugs(AEDs)and become refractory to treatment,so more effective treatment options need to be explored.The syndrome element is the basic diagnostic unit of Chinese medicine,and it helps to understand the syndrome rules and mechanism of the disease and guides the clinical treatment of the disease.To investigate the distribution of syndrome element in refractory epilepsy has important clinical significance for the future prevention and treatment of refractory epilepsy in Chinese medicine.Chaibei Zhixian decoction is an anti-epileptic formula to treatment refractory epilepsy.It has been shown to reduce the frequency of seizures and improve the quality of life of patients with intractable epilepsy.However,it is not known whether Chaibei Zhixian decoction can exert neuroprotective effects by modulating the nuclear factor E2-related factor 2(Nrf2)pathway and the Nod-like receptor protein 3(NLRP3)inflammatory vesicle pathway.Objective:A retrospective study of the case data of patients with refractory epilepsy was conducted to explore the distribution characteristics of their syndrome element and to provide ideas for the treatment of refractory epilepsy.A meta-analysis was conducted to objectively evaluate the clinical efficacy and safety of the method of resolving phlegm and relieving depression in refractory epilepsy.The effect of Chaibei Zhixian decoction on oxidative stress and inflammatory response in refractory epilepsy was explored through clinical research,and the effect and mechanism of Chaibei Zhixian decoction on oxidative stress and inflammatory response in refractory epilepsy was investigated using the animal model of epileptogenesis with kainic acid to provide a basis for its clinical application.Methods:1.The data collected from 236 cases of patients with refractory epilepsy were entered using Excel and standardised for symptom terminology.With reference to the syndrome element scale,the disease location and disease nature of each symptom were identified,the weight of the syndrome element was calculated and the diagnosis of the syndrome element was discriminated.The Ancient and Modern Medical Case Cloud Platform V2.2.3 data mining module was used for statistical analysis,association rules,hierarchical clustering and complex network analysis of case data.SPSS 25.0 software was used to cluster and analyse the syndrome elements of the case data.2.The randomized controlled trials on the treatment of refractory epilepsy by combining anti-epileptic drugs(AEDs)with the method of resolving phlegm and relieving depression were searched at home and abroad.Duplicate literature was searched and deleted by the literature management software.The remaining literature was screened according to the inclusion and exclusion criteria.The final included literature was extracted and evaluated for quality,and the data collected were analyzed using Revman 5.4.1 software and Stata 15.1 software.The primary outcome indicators were seizure frequency,seizure duration,and clinical efficiency,and the secondary outcome indicators were the MoCA scale score,EEG abnormality rate,quality of life scale,self-rating depressive scale and safety.3.Wistar rats were randomly divided into sham and model groups.The model group was established by localized injection of kainic acid(KA)into the lateral ventricle and then randomly divided into Chinese medicine and model groups.Rats in the sham group were given 3 μL of saline in the same way.Rats in the Chinese medicine group were given Chaibei Zhixian decoction.The rats in the sham-operated and model groups were given an equal volume of 0.9%NaCl solution.After 8 weeks of continuous intervention,the rats were observed for general conditions.Morris water maze test,and histopathological changes in the hippocampus by HE staining.ELISA kits were used to measure serum MDA,superoxide dismutase(SOD)and catalase(CAT)activity,interleukin-1β(IL-1β)and interleukin-18(IL-18)levels,and Western Blot was used to detect the expression of Kelch-like ECH-associated protein1(Keapl),Nrf2,heme oxygenase-1(HO-1).NLRP3 and Caspase-1.Results:1.Analysis of clinical data from patients with refractory epilepsy revealed a total of 120 symptoms in the seizure phase,of which the top 10 symptoms were confusion,twitching of limbs,gaze fixedly,rigidity of limbs,salivation at the mouth,freeze of mind,tightly closed teeth,rigidity of the neck,strange screams,and repetitive hand movements.There are 159 resting symptoms,with the top 10 symptoms being weakness,forgetfulness,depression,difficulty sleeping,impetuous,palpitations,dizziness,irritability,headache and wakefulness.There are 22 types of tongue texture,with the top 5 being pale red tongue,pale dark tongue,red tongue,red tip of tongue,and dark red tongue.There are 12 types of tongue coating,the top 5 being greasy coating,white coating,thin coating,yellow coating and little coating.There are 13 types of pulse diagnosis,the first 5 being smooth pulse,thin pulse,stringent pulse,sunken pulse,and fast pulse.Syndrome differentiation was used to identify the syndromes in patients with refractory epilepsy.A total of 13 types of diseased region elements were found in patients with refractory epilepsy,the more common ones being mind(brain),liver,kidney,heart,spleen,and,stomach.A total of 19 types of pathogenic syndrome elements were found,the more common ones being qi blockage,phlegm,moving wind,yin deficiency,dampness,qi deficiency,fire,blood deficiency,qi stagnation,yang hyperactivity,yang deficiency,and damage of essence.2.The results showed that the method significantly reduced the monthly seizure frequency(MD=-2.00,95%CI-2.80 to-1.20,P<0.01),improved EEG abnormality rates(OR=0.22,95%CI 0.08 to 0.59,P<0.01),improved quality of life(MD=8.35,95%CI 5.10 to 11.59,P<0.01),the MoCA scale score(MD=4.20,95%CI 2.14 to 6.26,P<0.01),self-rating depressive scale(MD=-6.38,95%CI-9.75 to-3.01,P<0.01),and its overall effectiveness was also better than that of AEDs alone(OR=4.31,95%CI 2.86 to 6.49,P<0.01).In terms of safety,no significant difference was shown between the combination of AEDs and Chinese medicine and AEDs alone(OR=0.75,95%CI 0.45 to 1.27,P>0.05).3.The results showed that compared with the model group,the TCM group could significantly reduce the frequency of weekly seizures in rats at 4 weeks(P<0.01)and 8 weeks(P<0.01).On days 1 to 5 of the positioning voyage in Morris water maze,the escape latency was significantly longer in the model group compared to the sham-operated group(P<0.05).There was no significant difference between the TCM group and the model group on day 1(P>0.05).On days 2 to 5 of the localization voyage,the escape latency of the rats in the TCM group was significantly shorter compared with that of the model group(P<0.05).In the spatial exploration experiment,the total distance travelled by the rats in the model group was significantly longer compared with the sham-operated group(P<0.05),and the distance travelled in the platform area,the number of times crossing the platform and the residence time in the platform quadrant were all significantly reduced(P<0.05).Compared with the model group,the total distance travelled by the rats in the Chinese medicine group was significantly shorter(P<0.05),and the distance travelled in the platform area,the number of times crossing the platform and the residence time in the platform quadrant were all significantly increased(P<0.05).HE staining showed that the brain tissues of the rats in the model group were significantly abnormal,with a large number of neuronal degeneration in the CA1,CA3 and DG areas of the hippocampus,with deep staining.In the Chinese medicine group,a small amount of neuronal degeneration,deep staining were observed.Compared with the sham-operated group,the serum SOD and CAT activities the model group were significantly lower(P<0.01,P<0.05),MDA levels were significantly increased(P<0.001),the levels of IL-1β and IL-18 were significantly increased(P<0.001,P<0.001).Compared with the model group,serum SOD and CAT activities in TCM group were significantly higher(P<0.05,P<0.05),MDA levels were significantly lower(P<0.01),and serum IL-1β and IL-18 levels were significantly lower(P<0.01,P<0.01).Western blot results showed that,compared with the sham-operated group,the hippocampal tissues of rats in the model group Keap1 protein expression level was significantly increased(P<0.001),Nrf2 and HO-1 protein expression was significantly decreased(P<0.001,P<0.001),NLRP3 and Caspase-1 protein expression level was significantly increased(P<0.001,P<0.001).Compared with the model group,the hippocampal tissues of rats in the TCM group showed significantly lower expression of Keap1(P<0.001).significantly higher expression of Nrf2 and HO-1(P<0.01,P<0.01),and significantly lower expression of NLRP3 and Caspase-1(P<0.01,P<0.05).Conclusion:1.According to the distribution characteristics of syndrome elements in patients with refractory epilepsy,it is suggested that the treatment of refractory epilepsy should mainly focus on calming endogenous wind.resuscitation,and dissolving depression and phlegm,at the same time should also pay attention to the existence of Yin deficiency,fire(heat),blood deficiency,qi deficiency,Yang deficiency,damage of essence and other factors.2.The method of relieving depression and resolving phlegm can significantly reduce the frequency of seizure,improve the abnormal rate of EEG,the quality of life,cognitive function,depressive state,and its overall effective rate is also better than that of AEDs alone.3.Chaibei Zhixian Decoction can reduce the level of oxide and increase the level of antioxidant enzymes by regulating Nrf2/Keapl/HO-1 signaling pathway,and at the same time,it can also inhibit the activation of inflammatory corpuscles of NLRP3,reduce the levels of inflammatory factors of IL-1β and IL-18,to decrease the frequency of seizure,exert neuroprotective effects,and improve the learning and memory ability of epileptic rats induced by kainic acid.