Correlation Analysis Of Blood Pressure And Plasma Cytokines In CACNA1D Gene C.920A>G Mutant SD Rats

Objective: CACNA1 D exon c.920A>G mutant SD rats were constructed by genetic engineering.Blood pressure and plasma cytokines related to Th1,Th2,Th17 cells of the rats were observed,and analyse the correlation.Methods: Obtaining the CACNA1 D exon c.920A>G mutant rats by CRISPR-Cas9 gene editing technique.Blood pressure,heart rate and body weight of the rats were measured at 8,9,10,17,21,26,28 and 32 weeks of the age.Enzyme linked immunosorbent assay(ELISA)was used to detect plasma IL-2,IFN-?,IL-4,IL-5,IL-6,IL-10 and IL-17 A,and IFN-?/IL-4 were calculated.The relationship between blood pressure and IL-2,IFN-?,IL-4,IL-5,IL-6,IL-10,IL-17 A,IFN-?/IL-4 in rats was analysed.Results:(1)Based on gene sequencing,32 AA genotype(wild type)SD rats(male 13,female 19),64 AG genotype(heterozygous mutation)rats(male 26,female 38)and 37 GG genotype(homozygous mutation)rats(male 19,female 18)were successfully constructed.Using the random number table method,12 AA genotype SD rats(6 males,6 females),18 AG genotype SD rats(9 males,9 females)and 12 GG genotype SD rats(6 males,6 females)were randomly selected as the research objects.(2)At 32 weeks,SBP of male AG genotype rats(141.78± 12.48 mm Hg)were higher than that of male AA genotype rats(122.00± 13.46 mm Hg),P < 0.01.SBP of male GG genotype rats(141.67± 9.95 mm Hg)were higher than that of male AA genotype rats,P < 0.05.At 32 weeks,SBP of female AG genotype rats(142.89± 13.02 mm Hg)were higher than that of female AA genotype rats(121.67±15.35 mm Hg),P < 0.01.SBP of female GG genotype rats(138.00±6.57 mm Hg)were higher than that of female AA genotype rats,P < 0.05.(3)Compared with AA genotype male rats(1627.55± 230.18 pg /ml),AG genotype male rats(2029.00± 174.72 pg /ml)and GG genotype male rats(2020.99± 243.79 pg /ml)had higher IFN-?(P < 0.01).Compared with AA genotype male rats(13.51±1.96),IFN-?/IL-4 in AG genotype male rats(17.31±3.16)was higher,P < 0.01.There was no significant difference in plasma IL-2,IL-4,IL-5,IL-6,IL-10 and IL-17 A among AA,AG and GG genotypes of male rats.There was no statistical difference in plasma IL-2,IFN-?,IL-4,IFN-?/IL-4,IL-5,IL-6,IL-10 and IL-17 A among AA,AG and GG genotypes of female rats.(4)The rats were divided into hypertensive rats and normal blood pressure rats using SBP ? 140 mm Hg as the standard of hypertensive rats.IFN-? in hypertensive rats(2006.78± 233.45 pg/ml)was higher than that in normal blood pressure rats(1827.88±216.95 pg/ml,P < 0.05).IFN-?/IL-4 in hypertensive rats(17.59± 2.93)was higher than that in normal blood pressure rats(15.25±2.06,P < 0.01).There was no significant difference in plasma IL-2,IL-4,IL-5,IL-6,IL-10 and IL-17 A between the hypertensive rats and the normal blood pressure rats.(5)Binary Logistic regression analysis indicated that the exon of CACNA1 D gene C.920A>G mutation was an independent risk factor for hypertension(OR: 6.679,95%CI: 1.109-40.225,P=0.038).Conclusion:(1)Exon of CACNA1 D gene C.920A>G mutation is an independent risk factor for hypertension.(2)The occurrence of hypertension is associated with Th1/Th2 cell imbalance in exon CACNA1 D c.920A>G mutant rats,the specific mechanism needs to be further studied.