Experimental And Clinical Studies On The TIA And Mechanism Of Ischemia Tolerance By HuazhuoHeXue Granule

Objective:To observe the therapeutic effect of Huazhuo He Xue Granules on patients with TIA and the effect of NF-κB p65 signal protein expression.To explore the possible regulatory mechanism of Huazhuo Hexue Granules in the treatment of TIA and cerebral ischemic tolerance.Methods:1.Clinical study:72 patients(from the outpatient department and ward of Neurology Department of the Second Affiliated Hospital of Shandong Traditional Chinese Medicine University)who met the criteria were selected and randomly divided into control group(36 cases)and treatment group(36 cases).Both groups were treated with conventional western medicine,and the treatment group was added with Huazhuo He Xue granules.Both groups were treated continuously for two weeks,and NF-κB p65 level was detected with Elisa reagent box before and after treatment,and the patients were followed up for 90 days after treatment.2.Experimental research part:(1)Rat pheochromocytoma cells(PC12)were cultured,subcultured and stored in a refrigerator at-80°C for standby.(2)PC12 cells in logarithmic growth phase cultured in vitro were randomly divided into six groups:blank serum control group(Control,Con),Huazhuo He Xue granules drug-containing serum experimental group(Transient H,TH),blank serum experimental group(Transient K,TK),Huazhuo He Xue granules drug-containing serum model group(Model H,MH),blank serum model group(Model KMK),Huazhuo He Xue granules drug-containing serum EX527 model group(EXH).The control group did not receive any treatment.The experimental group received OGD 10 min simulated ischemic preconditioning(IP),cultured for 24 h,and then OGD 4h.The model group was directly treated with OGD for 4h after 24 h culture.(3)Cell survival rate was detected by CCK8 method,and NF-κB p65 protein expression was detected by immunofluorescence method.Results:1.The total effective rate of the treatment group was significantly higher than that of the control group(P < 0.05);The improvement of TCM syndromes in the treatment group was significantly better than that in the control group(P<0.05);The protein expression of NF-κB in serum of treatment group was significantly lower than that of control group(P<0.05).2.Cell survival rate: Con>TH>TK>MK(P<0.05);Con>TH>MH>EXH(P<0.05);Con>TH>MH>MK(P<0.05).3.NF-κB p65 protein expression: MK>MH>TH>Con(P<0.05);MK>TK>TH>Con(P<0.05);EXH>MH>TH>Con(P<0.05).Conclusion:1.Huazhuo Hexue Granules can significantly improve the clinical symptoms of TIA patients,and can effectively reduce the oxidative stress and inflammatory reaction of TIA patients.2.The medicated serum containing Huazhuo Hexue Granules after IP can enhance cell viability,effectively reduce the damage caused by OGD,and inhibit NF-κB p65 protein expression.According to clinical and experimental studies,By inhibiting NF-κB protein level,Huazhuo Hexue Granules can inhibit oxidative stress and reduce inflammatory reaction,thus obtaining the effect of brain protection.