The Effect Of Zhuanggu Zhitong Recipe Containing Serum On The Balance Of CD4+CD25+Treg/Th17 In Spleen Lymphocytes Of BALB/c Mice In Vitro

Objectives:To investigate the effects of Zhuanggu Zhitong Capsule(ZGZT capsule)on the balance of CD4+CD25+Treg/Th 17 through the study on the influence of ZGZT capsule on the expression of CD4+CD25+Treg,Th17 related cytokines,specific RNA and protein in mouse spleen lymphocytes cultivated in vitro.Methods:Preparation of medicine-containing serum:osteopo-rosis models were established by removing ovaries of female rats,and the bloods of the mice were collected and their serums were separated before they were divided into different groups for gavage for 3 months.Spleen lymphocytes of BALB/c mouse were prepared and were divided into 6 groups:the sham operation group,the model group,the estradiol valerate positive control group,the high-dose ZGZT Capsule group,the medium-dose ZGZT Capsule group and the low-dose ZGZT Capsule group.Each group was cultivated in 6 wells for 72h,and ELISA was adopted and culture medium collected to investigate the effect of ZGZT Capsule on TGF-β and IL-17 contents,and RT-PCR was adopted and cells collected to investigate the effects of ZGZT Capsule on the expression of the RNA of Foxp3 and RORγt RNA,and Western-blot was applied to detect the the effect of ZGZT Capsule on the expression of Foxp3 and RORγt proteins.Results:1.The differences between the model group with the estradiol valerate positive control group,the low-dose ZGZT Capsule group,the medium-dose ZGZT Capsule group and the high-dose ZGZT Capsule group in TGF-β concentration in the supernatant of cell culture media were statistically significant or highly statistically significant(P<0.05 or P<0.01);IL-17 concentration in the supernatant of cell culture media declined,especially of the medium-dose ZGZT Capsule group and the estradiol valerate positive control group,the difference being significant(P<0.05).2.Compared with the model group,the expression of Foxp3 mRNA in each medicine-containing serum group increased.In addition to the low-dose ZGZTCapsule group(P>0.05),the differences between the model group and the resting groups were statistically significant(P<0.05);the expression of RORyt RNA declined,the difference being statistically significant or highly statistically significant(P<0.05 or P<0.01);3.Compared with the model group,the expressions of Foxp3 and RORyt proteins in other groups were statistically significant(P<0.05).Conclusion:1.Compared with the model group,TGF-β concentrations in the sup-ernatant of cell culture media of estradiol valerate positive control group,the low-dose ZGZT Capsule group,the medium-dose ZGZT Capsule group and the high-dose ZGZT Capsule group increase,presenting positive correlation with the dose;the concentration of IL-17 decreases,and ZGZT Capsule regulates TGF-β concentration significantly;2.From the perspective of RNA and protein,it can be proved that ZGZT Cap-sule greatly affects the proportion of Treg/Th17,and significantly improves the im-balance of Treg/Th17 cells in the model group,and its dose is positively correlated with the concentration of Treg.