| 1、Effect of Hanjing Decoction on Calcitonin Gene-related Peptide and Relaxin in Serum of Rats with Cold-Coagulation HypertensionObjective: To study the effect of Hanjing Decoction on calcitonin gene-related peptide and relaxin in serum of hypertensive rats with cold coagulation,and to explore the mechanism of reducing hypertension by Hanjing Decoction.Methods: Twenty-four SD rats were randomly divided into control group,model group and Hanjing Decoction group,and fed for one week.One week later,the model group and the Hanjing Decoction group were given high salt diet containing 8% and drinking water freely,and placed in(-15 +2)℃ refrigerator every day.The control group was fed with normal diet and free drinking water at 24~26 C.The blood pressure of rat tail artery was measured three times a week.From the seventh week,the Hanjing Decoction group was given traditional Chinese medicine by gastric lavage,while the model group was given normal saline by gastric lavage for two weeks.Two weeks later,blood samples were taken from the decapitated heads.Calcitonin gene-related peptide(CGRP)in serum was detected by radio immunoassay(RIA)and relaxin in serum was detected by enzyme-linked immunosorbent assay(ELISA).Results: 1.Serum CGRP(calcitonin generelated peptide):Compared with the normal group,the expression level of CGRP in the model group was lower,P(27)0.05.Compared with the model group,the expression level of CGRP in the Hanjing Decoction group was higher,P(27)0.05.2.Serum RLX: Compared with the normal group,the serum RLX in the model group was lower,with significant difference,P(27)0.05.Compared with the model group,the serum RLX expression level in the Hanjing Decoction group was higher,with significant difference,P(27)0.05.Conclusions: 1.The hypotensive effect of Hanjing Decoction may be related to the elevation of serum calcitonin gene-related peptide level.2.The hypotensive effect of Hanjing Decoction may be related to the elevation of serum relaxin level.2、Effects of Hanjing Decoction-containing Serum on UCP2 and RXFP2 of Vascular Smooth Muscle Cells in Rats Induced by Cold StimulationObjective:By culturing vascular smooth muscle cells of SD rats in vitro and giving cold stimulation,the effects of serum containing Hanjing Decoction on UCP2 and RXFP2 of rat smooth muscle cells stimulated by cold were studied,and the prevention and treatment mechanism of Hanjing Decoction on cold-induced hypertension was discussed,which provided experimental basis for clinical prevention and treatment of atherosclerosis,hypertension and other related cardiovascular diseases by Hanjing decoction.Methods: Rat vascular smooth muscle cell line(A7R5)was selected and subcultured.Smooth muscle cells of 4-6 generations were taken for experiment.The cold stimulation time was 2h,4h,6h and 8h,respectively.Cell viability was counted by trypan blue staining,OD value was detected by MTT method,and the optimal cold stimulation time was selected.Twenty male SD rats were randomly divided into five groups.The serum containing Chinese herbs was prepared by intragastric administration of Chinese herbs and normal saline,respectively,and the smooth muscle cells of different groups were treated.The smooth muscle cells were randomly divided into normal group,model group,Hanjing Decoction group,Zhijing group and Maxinfu group.The original culture medium was inhaled and PBS was washed once.The drug-containing serum was added into 1.Normal control group: DMEM + 10% normal rat serum;2.Model group: DMEM + 10%model rat serum 3.Hanjing Decoction group: DMEM + 10% Hanjing Decoction rat serum;4.Zhijing group: DMEM + 10% Zhijing powder rat serum;5.Maxinfu group: DMEM + 10% Mahuangfuzi Xixin Decoction rat serum.Each group was cultured in 10% serum cell incubator for 24 hours.After the drug-containing serum was cultured,the model group,Hanjing Decoction group,Zhijing group and Maxinfu group were given cold stimulation in a 20℃incubator,while the normal group was cultured in a37℃incubator.The expression of uncoupling protein 2 and relaxin receptor 2in smooth muscle cells was detected by fluorescence quantitative PCR and Western-blotting.Results:1.When the temperature of cold stimulation was set at 20℃,the survival rate of cold stimulation time decreased at 4 h,and there was significant difference between 4 h group and 6 h group,P(27)0.05.2.The expression of uncoupling protein 2 in vascular smooth muscle cells was significantly lower than that in normal group,P(27)0.05.Compared with model group,the expression of uncoupling protein 2 in Hanjing Decoction group was higher than that in Maxinfu group and Zhijing group,P(27)0.05 or 0.01.Compared with Hanjing Decoction group,the expression of protein in Zhijing group decreased,P(27)0.05.Compared with the normal group,the expression of relaxin receptor 2 protein in vascular smooth muscle cells decreased,P(27)0.01.Compared with the model group,the expression of protein in Hanjing Decoction group,Maxinfu group and Zhijing group increased,P(27)0.05 or 0,01.Compared with Hanjing Decoction group,the expression of Maxinfu group decreased P(27)0.05.3.The expression of uncoupling protein 2 gene in vascular smooth muscle cells was lower in model group than in normal group,P(27)0.05.Compared with model group,the expression of uncoupling protein 2 gene in Hanjing Decoction group and Maxinfu group was higher,P(27)0.05 or 0.01.Compared with Hanjing Decoction group,the expression of uncoupling protein 2 gene in Zhijing group was lower,P(27)0.05.Compared with the normal group,the gene expression of relaxin receptor 2 in vascular smooth muscle cells in model group decreased,P(27)0.05.Compared with model group,the gene expression in Hanjing Decoction group,Maxinfu group and Zhijing group increased,P(27)0.05 or 0.01.Compared with Hanjing Decoction group,the gene expression in Maxinfu group and Zhijing group decreased,P(27)0.05.Conclusions: 1.Cold stimulation can reduce the expression of uncoupling protein 2 and relaxin receptor 2 in vascular smooth muscle cells.2.Hanjing Decoction can increase the expression of uncoupling protein 2 and relaxin receptor 2 in vascular smooth muscle cells stimulated by cold.3.Hanjing Decoction is better than Mahuang Xixin Fuzi Decoction and Zhijing Powder in increasing uncoupling protein 2 and relaxin receptor 2 of vascular smooth muscle cells. |
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