| Objective Maternal obesity leads to many gestational complications and adverse fetal birth outcomes with immediate and long-term consequences.At the same time,maternal obesity is associated with increasing inflammatory levels in the maternal circulation and increased placental inflammatory status.Docosahexaenoic acid(DHA)has a significant anti-inflammatory effect that regulates levels of inflammatory factor by itself and its metabolites.In this experiment,after establishing an obesity model and successfully conceiving,different DHA treatments were performed.To verify the effect on anti-inflammatory of DHA during pregnancy to regulate NF-κB signaling pathway and NLRP3 inflammsome,we compared the pathological changes of maternal and placental tissues,detected levels of inflammation in plasma and placental tissues.And verify the regulation of inflammatory bodies can be mediated through the GPR120 receptor.Methods Four-week-old C57BL/6J females were divided into normal control group(CON)and high fat diet feeding group(HFD)according to the principle of complete randomization.After 9 weeks of feeding,the HFD group screened into the obese group according to the weight status.All the mice were caged at 1:2(♂:♀).After successful conception,the control group continued to feed the normal diet;the obese group was randomly divided into four groups including the obesity model(HCD)group,the low-dose DHA intervention(HCD-LD)group(52 mg/kg,calculated as DHA),and the medium-dose DHA intervention(HCD-MD).group(104 mg/kg),and high dose DHA intervention(HCD-HD group)group(208 mg/kg)..Different doses of DHA,were given with high fat diet.During the feeding period,the food intake and body weight of each group of mice were weighed regularly.Mice were fasted at 8:00 pm and the water was not fasted.After measuring the fasting blood glucose on the morning of GD18.Pregnant mice were anesthetized by intraperitoneal injection of 10% chloral hydrate,and then collected the plasma to determine the inflammatory factors.The liver was dissected and weighed,and part of the liver was embedded with OTC and fixed with 4%polyformaldehyde.The placenta was removed and weighed,and pick some fixed with4% paraformaldehyde and the other were stored for real-time quantification PCR and Western blotting.Results The weight gain during pregnancy in the HCD group and each dose group was higher than that in the control group(p<0.01).There was no significant difference in the weight gain of pregnant rats during the middle,late and whole pregnancy(p>0.05).Compared with the control group,The liver weight of the HCD group was significantly increased(p<0.01).There was no significant difference between the dose groups and the HCD group(p>0.05).The liver tissue of the HCD group showed a large amount of lipid droplet accumulation and inflammation.Sex cells,HCD-HD group was significantly reduced after DHA intervention,and the liver tissue structure tends to be normal.Oil red O staining sections showed that the control group had some colored lipid droplets,while the HCD group showed significant red staining,and the lipid droplets in the middle and high dose groups were significantly reduced.The levels of plasma IL-1β and TNF-α in the HCD group were significantly higher than those in the CON group.The levels of IL-1β in the middle and high dose groups were significantly lower(P<0.05,P<0.01),and the TNF-α levels were lower in the high dose group.<0.01).The body weight and carcass ratio(fetal weight/placental weight)of the HCD group were significantly lower(P<0.01),and the middle and high dose groups were significantly higher than the HCD group(P<0.01).The placenta diameter of the different groups was not obvious.difference.Compared with the CON group,the TG,TC,and LDL levels in the HCD group were higher than those in the control group(P <0.05),and the HDL levels were lower(P < 0.05).Compared with the HCD group,the TC levels were lower in the middle and high dose groups(P < 0.01),LDL levels in the HCD-HD group decreased,TG levels in the HCD-LD group began to decrease(P<0.05),HDL levels were not significantly different(P> 0.05).HE staining of placental tissue showed that the placenta hyperemia in the HCD group was obvious and there was a large amount of inflammatory cell infiltration,and the HCD-HD group improved significantly.The m RNA and protein levels of CD36,FATP-4 and FABP-pm in HCD group were significantly inhibited(p<0.01),and the level of CD36 m RNA in HCD-HD group was significantly up-regulated(p<0.01).The level of FATP-4 m RNA in each dose group increased.,but the difference was not statistically significant;FABP-MD group began to appear up-regulated in HCD-MD group(p<0.01);compared with HCD group,HCD-LD group CD36,FABP-pm(p<0.01),medium and high dose The expression of FATP-4 protein was up-regulated significantly(p<0.05).The expression of GPR120 gene and protein in placenta of pregnant rats in HCD group did not change significantly.Compared with HCD group,the m RNA level of HCD-HD group was significantly increased(P<0.05),and the protein expression of HCD-LD group was up-regulated(P<0.05).The NF-κB,IKK and IκB m RNA in the HCD group were significantly higher than those in the CON group(P<0.01).The NF-κB and IκB in the HCD-LD group began to decrease,and the IKK HCD-MD group began to decrease(P<0.05).The expression of p-IκB protein in NF-κB and total protein was significantly increased in HCD group,and NF-κB in HCD-HD group was significantly decreased(P<0.01).P-IκB in HCD-MD group was significantly decreased(P<0.01).).The expression of total protein IκB(P<0.01)and cytoplasmic NF-κB(P<0.05)was decreased in HCD group,and the expression of medium and high dose group was up-regulated.The expressions of NLRP3 and Caspase-1 m RNA and protein were increased in HCD group(P<0.01).The expression of Caspase-1 gene in HCD-LD group and NLRP3 and middle and high dose groups were down-regulated(P<0.01).The expression of NLRP3 protein decreased with the increase of DHA intervention concentration(P<0.01),and the expression level of Caspase-1 protein was significantly decreased in HCD-HD group.IL-1β(P<0.01),TNF-α(P<0.05)and IL-6(P<0.01)m RNA levels were significantly up-regulated in HCD group.Compared with HCD group,HCD-LD group IL-1β,HCD IL-6 m RNA levels in the MD group began to decrease(P<0.01);TNF-α m RNA levels decreased in each treatment group,but the difference was not significant(P>0.05);IL-1β and TNF-α protein in HCD group The expression of IL-1β protein decreased in each dose group(P>0.05).The expression of TNF-α protein in the middle and high dose groups decreased significantly(P<0.01).Conclusion Increasing proinflammatory cytokines in maternal and fetal circulation with placental inflammation and other pathological changes in obese pregnant mice.DHA intervention during pregnancy can decrease the level of placental inflammation and affect the birth outcome of offspring.The possible pathway of regulation maybe DHA inhibited the activation of NLRP3 inflammasome and NF-κB signaling pathways to inhibit the elevation of pro-inflammatory cytokines by activating GPR120 receptor. |
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