The Study On Immunoregulatory Mechanism Of Polysaccharides Of Pharbitis Semen In Treatment Of Membranous Nephropathy In Rats

Objective:Semen Pharbitidis is the dry and mature seed of Pharbitis nil(L.)Choisy or Pharbitis purpurea(L.)Voigt belonged to the convolvulaceae family.It is cold in nature and bitter in flavor.According to the previous study of our research group,Semen Pharbitidis were separated into four components:resin glycosides,phenolic acids,fatty oils and polysaccharides.And it has been proved by experiment that Semen Pharbitidis have the therapeutic effect on membranous nephropathy in rats.The effective site may be a polysaccharide component.In this article,membranous nephropathy in rats induced by cationized bovine serum albumin(C-BSA).The immune regulation mechanism of Semen Pharbitidis polysaccharide treatment of membranous nephropathy rats was explained.Methods:In this experiment,60 male and healthy SD rats were randomly divided into six groups:normal group,model group,positive group,low-dose group of Semen Pharbitidis polysaccharide,medium-dose group of Semen Pharbitidis polysaccharide,and high-dose group of Semen Pharbitidis polysaccharide,10 in each group.Intragastric administration of low,medium,and high doses of Semen Pharbitidis polysaccharide was used.And rats in the positive drug group were given intragastric administration of Tripterygium glycoside tablets,while the blank and model groups were given intragastric administration of normal saline for a total of four weeks.The pharmacodynamic evaluation of rats with membranous nephropathy was studied from the aspects of 24h urinary protein,kidney index,serum biochemistry,antioxidant index,and optical examination of kidneys.The levels of nuclear factor NF-κB and inflammatory cytokines TNF-α,IL-1β,IL-6,IL-8 and TGF-β1 in serum were detected by ELISA.And immunohistochemistry and RT-PCR were used to detect the protein expression and mRNA expression of the target protein in the kidney.Results:The pharmacodynamic evaluation results showed that the rats in the low,medium and high dose groups were significantly relieved compared to the model group after the administration of Semen Pharbitidis polysaccharide polysaccharides,24h urine protein content and kidney index were decreased significantly,TG,TC,Scr,BUN in serum content decreased significantly,TP and ALB in serum increased significantly,MDA decreased significantly,SOD and GSH-PX activity increased significantly.Observed from the results of light microscopy,compared with the model group,the glomerular bifurcation,tubular swelling and fibrosis of the rats in the low,medium,and high groups of Semen Pharbitidis polysaccharides were significantly improved.The immunomodulatory mechanism study showed that compared with the model group,The expression levels of the nuclear transcription factor NF-κB and inflammatory cytokines IL-1β,IL-6,IL-8,TNF-α and TGF-β1 mRNA in the renal tissues of the rats in the low,medium,and high groups of Semen Pharbitidis polysaccharides were significantly downregulated,the protein expression of the nuclear transcription factor NF-κB and inflammatory cytokines IL-1β,IL-6,IL-8,TNF-α,TGF-β1 in serum and renal tissues were significantly decreased.Conclusion:The experiment showed that the polysaccharides of Semen Pharbitidis had a good therapeutic effect on membranous nephropathy rats.The mechanism is mainly to regulate gene transcription and protein expression of the nuclear factor NF-κB and inflammatory cytokines IL-1β,IL-6,IL-8,TNF-α,TGF-β1 in serum and renal tissues.In addition,it can increase the antioxidant effect of the body and achieve the immune regulation effect on the rats with membranous nephropathy.