| Purpose : Reveal Qijishenkang Decoction in the prevention and treatment of Ig A Nephropathy(Ig AN),and explore the effects of Qijishenkang Decoction on Angiotensinⅱ(Angⅱ)inducing the proliferation of Human Mesangial cells(HMC),To observe the effects of Connective Tissue Growth Factor(CTGF)on JAK2/STAT3 signaling pathway and Connective Tissue Growth Factor.It can provide theoretical basis and prevention target for the treatment of Ig A nephropathy with traditional Chinese medicine.Material and method:1.Preparation of drug-containing serum:forty male Sprague-Dawley(SD)rats have been randomly divided into 5 businesses in accordance to physique weight,specifically everyday group,Western medication group,Qijishenkang Decoction low-dose group,Qijishenkang Decoction medium-dose group,and Qijishenkang Decoction high-dose group.The dosage was once calculated.After non-stop ingastric administration for one week,blood samples have been accumulated from stomach aorta and amassed in groups.The accrued blood was once transferred to a centrifuge for centrifugation.At the quit of centrifugation,the supernatant was once taken and positioned in a 56℃ water tub for 30 min for inactivation treatment,then filtered,sterilized,and separated for use.2.Culture,proliferation and grouping of mesangial cells: Human glomerular mesangial cells had been inoculated into way of life flask,then delivered into RPMI-1640 medium and transferred into an incubator;Proliferation of mesangial cells was once brought about by using Angⅱ.According to the experimental scheme,the cells have been divided into :(1)regular team(2)mannequin crew(3)western medication crew(4)low dose crew of Qijishenkang Decoction(5)medium dose team of Qijishenkang Decoction(6)excessive dose crew of Qijishenkang Decoction.3.Detection methods :(1)MTT assay was used to detect cell proliferation;(2)The contents of p JAK2,STAT3 and CTGF in mesangial cells were detected by enzyme-linked immunosorbent assay(ELISA)(3)the expressions of STAT3 m RNA,PIAS3 m RNA and CTGFm RNA were detected by real-time fluorescence quantitative PCR.Results:1.MTT assay confirmed that the proliferation of mesangial cells in the mannequin team was once extensively greater than that in the regular crew(P<0.01),indicating that Ang ⅱ should stimulate the proliferation of HMC and the modeling used to be successful.Compared with mannequin group,the proliferation of mesangial cells in western remedy group,Qijishenkang decoction medium-dose and high-dose organizations had considerable distinction(P<0.01),however the proliferation of mesangial cells in low-dose team had no huge distinction(P>0.05).2.ELISA results showed that:(1)p JAK2 expression: Compared with regular group,the expression of p JAK2 in mannequin crew was once extensively elevated(P<0.01);Compared with mannequin group,the effects of western medication crew and Qijishenkang Decoction had been extensively lowered(P<0.01).There was once a large distinction amongst exclusive dosage organizations of Qijishenkang Decoction and there used to be a dose-effect relationship(P<0.01).(2)STAT3 expression: Compared with regular group,the expression of STAT3 in mannequin crew was once extensively elevated(P<0.01);Compared with mannequin group,the effects of western medication crew and Qijishenkang Decoction had been extensively lowered(P<0.01).There was once a large distinction amongst exclusive dosage organizations of Qijishenkang Decoction and there used to be a dose-effect relationship(P<0.01).(3)CTGF expression: Compared with the regular group,the expression of CTGF in the mannequin crew used to be appreciably improved(P<0.01);Compared with mannequin group,the consequences of western medication crew and Qijishenkang Decoction have been extensively reduced(P<0.01).There have been giant variations amongst distinctive dose businesses of Qijishenkang Decoction,and there used to be a dose-effect relationship amongst exclusive dose agencies(P<0.05).3.The results of real-time quantitative PCR showed that:(1)STAT3m RNA expression:Compared with regular group,STAT3 m RNA expression in mannequin team was once extensively extended(p<0.01);Compared with mannequin group,the consequences of western remedy team and Qijishenkang Decoction in every dose agencies had been extensively lowered(P <0.05).There used to be vast distinction between the low-dose team and the medium-dose and high-dose corporations(P<0.01),however no substantial distinction between the medium-dose and high-dose businesses(P>0.05).(2)PIAS3m RNA expression:Compared with everyday group,the expression of PIAS3 m RNA in mannequin crew was once considerably diminished(p<0.01);Compared with mannequin group,the effects of western medication crew and Qijishenkang Decoction dose companies had been substantially elevated(P<0.05).There used to be big distinction between the low-dose crew and the medium-dose and high-dose businesses(P<0.01),however no considerable distinction between the medium-dose and high-dose agencies(P>0.05).(3)CTGFm RNA expression:Compared with everyday group,CTGFm RNA expression in mannequin team was once extensively elevated(p<0.01);Compared with mannequin group,the effects of western medication team and Qijishenkang Decoction had been appreciably reduced(P <0.01).There was once great distinction between the low-dose team and the medium-dose and high-dose agencies(P<0.01),however no giant distinction between the medium-dose and high-dose businesses(P>0.05).Conclusion:1.Qijishenkang Decoction can inhibit the proliferation of human glomerular mesangial cells induced by Angⅱ.2.To clarify that the targets of Qijishenkang Decoction on inhibiting proliferation of human mesangial cells may be JAK2,STAT3,PIAS3 and CTGF.3.By inhibiting the expression of p JAK2 and STAT3 and promoting the expression of PIAS3,Qijishenkang Decoction inhibited the JAK2/STAT3 signaling pathway,reduced the content of CTGF in HMCs,inhibited the proliferation of HMCs,alleviated the process of renal fibrosis,changed the pathological state of Ig A nephropathy,and prevented the further deterioration of Ig AN.This may be one of the mechanisms of Qijishenkang Decoction in treating Ig A nephropathy. |
PDF Full Text Request