Effects of placental periodontal pathogens on preeclampsia outcomes

doi:10.3877/cma.j.issn.1673-5250.2015.02.013

Objectlve

To investigate the expressions of serum C reactive protein (CRP), four common periodontal pathogens, porphyromonas gingivalis (Pg), actinobacillus actinomycetemcomitans (An), fusobacterium nucleatum (Fn), campylobacter rectus (Cr) and the alteration of NO formation in the placenta tissue of preeclampsia (PE) combined with periodontitis.

Methods

A total of 25 pregnant women who visited Guangdong Women and Children Hospital from January 2012 to October 2013 were included in this study (PE group). Meanwhile, another 27 healthy pregnant women were collected into control group. PE group and control group were both further divided into two sub-groups respectivley, PE group combined with periodontitis (PE-A sub-group, n=16), PE sub-group combined with no periodontitis (PE-B sub-group, n=9), control group combined with periodontitis (control-A sub-group, n=17), and control group combined with no periodontitis (control-B sub-group, n=10). There were no significant differences on age ratio, body weight, and body mass index (BMI) among four groups (P>0.05). The study protocol was approved by the Ethical Review Board of Investigation in Human Being of Guangdong Women and Children Hospital. Informed consent was obtained from the parent of each participates. CRP was tested by immunological transmission turbidimetry. Four periodontal pathogens in the placenta tissues were tested by PCR. The levels of endothelial nitric oxide synthase (eNOS) and nitric oxide (NO) in placenta tissues were tested by Western blotting and Griess assay, respectively.

Results

①There were no significant differences on the incidence rate of periodontitis, PD, CAL and BOP between PE group and control group (P>0.05). ②There were significant differences on MSBP, MDBP, incidence rate of premature labor, and IUGR between PE-A sub-group and control-A sub-group, PE-A sub-group and control-B sub-group (MSBP: P<0.05; MDBP: P<0.05; incidence of premature delivery: P<0.05; IUGR: P<0.05). ③ The levels of MSBP, MDBP, incidence rate of premature labor, and IUGR between PE-B sub-group and control-A sub-group, PE-B sub-group and control-B sub-group (MSBP: t=50.5, 43.3; P<0.05; MDBP: t=13.3, 16.3; P<0.05; incidence of premature delivery: P<0.05; IUGR: P<0.05). ④ The incidence rate of fetal distress of PE-A sub-group was significant higher than that of control-B sub-group(P=0.018). ⑤ There were significant differences on gestational age, fetal birth weight, NO biosynthesis of placental tissues and placenta weight between PE-A sub-group and control-A sub-group, PE-A sub-group and control-B sub-group (gestational age: t=-7.0, -6.3; P<0.05; fetal birth weight: P<0.05; P<0.05; NO biosynthesis of placental tissues: t=-28.8, -42.1; P<0.05; placenta weight : P<0.05, P<0.001)). ⑥ There were significant differences on gestational age, fetal birth weight and placenta weight between PE-B sub-group and control-A sub-group, PE-B sub-group and control-B sub-group (gestational age: t=-5.9, -5.2, P<0.05; fetal birth weight: P<0.05; placenta weight: P<0.05, P<0.001 ). ⑦ The levels of serum CRP, incidence rates of Pg, Aa, and Cr in placental tissues of PE-A sub-groups were higher than those of control-B sub-group with significant differences (serum CRP: t=2.2, P=0.038; incidence rates of Pg, Aa, and Cr in placental tissues: χ²=5.097, P=0.024; χ²=7.659, P=0.006; P=0.044). ⑧ The b-actin/ eNOS value of PE-A sub-group was higher than that of PE-B sub-group, control-A sub-group and control-B sub-group, with significant differences (P<0.05). ⑨There were no significant differences on placental index among four groups (P>0.05).

Conclusions

Periodontal pathogens /byproducts may invade the placenta tissues and stimulate the inflammatory response against the activity of eNOS in placenta tissues. Then periodontal pathogens may lead to the exacerbation of PE adverse pregnancy outcomes.

Key words: Pre-eclampsia, Periodontitis, C-reactive protein, Nitric oxide synthase

表1 牙周细菌引物序列长度

Table 1 Sequences length of primer of periodental pathogens

牙周细菌	引物序列	扩增片段长度(bp)
Pg	正向引物5'-TGTAGATGACTGATGGTGAAAACC-3'	131
	反向引物5'-ACGTCATCCACACCTTCCTC-3'	?
Aa	正向引物5'-GCTAATACCGCCTAGAGTCGT-3'	235
	反向引物5'-ATTTCACACCTCACTTAAAGGT-3'	?
Fn	正向引物5'-GGCCACAAGGGGACTGAGACA-3'	185
?	反向引物5'-TTTAGCCGTCACTTCTTCTGTTGG-3'	?
Cr	正向引物5'-TTTCGGAGCGTAAACTCCTTT TC-3'	598
?	反向引物5'-TTTCTGCAAGCAGACACTCTT-3'	?

表1 牙周细菌引物序列长度

Table 1 Sequences length of primer of periodental pathogens

牙周细菌	引物序列	扩增片段长度(bp)
Pg	正向引物5'-TGTAGATGACTGATGGTGAAAACC-3'	131
	反向引物5'-ACGTCATCCACACCTTCCTC-3'	?
Aa	正向引物5'-GCTAATACCGCCTAGAGTCGT-3'	235
	反向引物5'-ATTTCACACCTCACTTAAAGGT-3'	?
Fn	正向引物5'-GGCCACAAGGGGACTGAGACA-3'	185
?	反向引物5'-TTTAGCCGTCACTTCTTCTGTTGG-3'	?
Cr	正向引物5'-TTTCGGAGCGTAAACTCCTTT TC-3'	598
?	反向引物5'-TTTCTGCAAGCAGACACTCTT-3'	?

表2 PE组和对照组孕妇牙周检查结果比较(±s)

Table 2 Comparison of periodontal examination between PE group and control group (±s)

组别	例数	牙周炎患病率例数(%)	PD(mm)	CAL(mm)	BOP例数(%)
PE组	25	16(64)	3.51±1.02	3.41±0.93	18(72.00)
对照组	27	10(37)	3.44±0.95	3.32±0.86	14(51.85)
t值/χ²值	?	3.775	0.980	0.158	2.23
P值	?	0.095	0.327	0.693	0.136

表2 PE组和对照组孕妇牙周检查结果比较(±s)

Table 2 Comparison of periodontal examination between PE group and control group (±s)

组别	例数	牙周炎患病率例数(%)	PD(mm)	CAL(mm)	BOP例数(%)
PE组	25	16(64)	3.51±1.02	3.41±0.93	18(72.00)
对照组	27	10(37)	3.44±0.95	3.32±0.86	14(51.85)
t值/χ²值	?	3.775	0.980	0.158	2.23
P值	?	0.095	0.327	0.693	0.136

表3 4组孕妇妊娠结局比较(±s)

Table 3 Comparison of pregnancy results among four groups (±s)

组别		例数	年龄(岁)	MSBP(mmHg)^a	MDBP (mmHg)^b	孕前BMI(kg/m²)	分娩孕龄(孕周)^c
PE组		?	?	?	?	?	?
?	PE-A亚组	16	28.0±4.4	156.2±21.7	95.6±16.8	23.4±3.3	32.9.±2.7
?	PE-B亚组	9	29.0±3.6	159.1±23.1	92.1± 9.2	23.1±2.6	33.9±2.5
对照组		?	?	?	?	?	?
?	对照-A亚组	10	25.0±3.6	108.1±13.1	78.1±9.2	21.3±4.2	39.9±0.8
?	对照-B亚组	17	26.0±7.2	116.1±12.4	75.7±7.8	21.6±4.4	39.1±0.9
t/χ²值		?	2.102	28.5	19.1	1.9	38.0
P值		?	0.112	0.000	0.000	0.139	0.000
组别		出生体质量(g)^d	胎儿宫内窘迫[例数(%)]^e	早产[例数(%)]^f	FGR[例数(%)]^g	生后1 min Apgar评分(分)	生后5 min Apgar评分(分)
PE组		?	?	?	?	?	?
?	PE-A亚组	2130±371	5(31)	13(81)	11(69)	8.6±0.7	9.8±0.4
?	PE-B亚组	2044±384	3 (8)	8(89)	5(56)	8.6±0.7	9.9±0.3
对照组		?	?	?	?	?	?
?	对照-A亚组	3184±308	1(10)	0 (0)	0 (0)	8.9±0.3	9.9±0.3
?	对照-B亚组	3284±358	0 (0)	0 (0)	0 (0)	9.0±0.0	10.0±0.0
t值/χ²值		44.7	10.1	48.4	32.0	7.7	1.1
P值		0.000	0.018	0.000	0.000	0.053	0.355

表3 4组孕妇妊娠结局比较(±s)

Table 3 Comparison of pregnancy results among four groups (±s)

组别		例数	年龄(岁)	MSBP(mmHg)^a	MDBP (mmHg)^b	孕前BMI(kg/m²)	分娩孕龄(孕周)^c
PE组		?	?	?	?	?	?
?	PE-A亚组	16	28.0±4.4	156.2±21.7	95.6±16.8	23.4±3.3	32.9.±2.7
?	PE-B亚组	9	29.0±3.6	159.1±23.1	92.1± 9.2	23.1±2.6	33.9±2.5
对照组		?	?	?	?	?	?
?	对照-A亚组	10	25.0±3.6	108.1±13.1	78.1±9.2	21.3±4.2	39.9±0.8
?	对照-B亚组	17	26.0±7.2	116.1±12.4	75.7±7.8	21.6±4.4	39.1±0.9
t/χ²值		?	2.102	28.5	19.1	1.9	38.0
P值		?	0.112	0.000	0.000	0.139	0.000
组别		出生体质量(g)^d	胎儿宫内窘迫[例数(%)]^e	早产[例数(%)]^f	FGR[例数(%)]^g	生后1 min Apgar评分(分)	生后5 min Apgar评分(分)
PE组		?	?	?	?	?	?
?	PE-A亚组	2130±371	5(31)	13(81)	11(69)	8.6±0.7	9.8±0.4
?	PE-B亚组	2044±384	3 (8)	8(89)	5(56)	8.6±0.7	9.9±0.3
对照组		?	?	?	?	?	?
?	对照-A亚组	3184±308	1(10)	0 (0)	0 (0)	8.9±0.3	9.9±0.3
?	对照-B亚组	3284±358	0 (0)	0 (0)	0 (0)	9.0±0.0	10.0±0.0
t值/χ²值		44.7	10.1	48.4	32.0	7.7	1.1
P值		0.000	0.018	0.000	0.000	0.053	0.355

表4 牙周菌在胎盘组织的分布及胎盘组织的NO含量情况

Table 4 The distribution of periodontal bacteria and the metabolism of NO in placenta

组别		例数	血清CRP^a	胎盘组织中检出牙周细菌^b				胎盘氧代谢β-actin/eNOS^c	NO(μmol/L)^d	胎盘系数	胎盘重量(g)^e
组别		例数	血清CRP^a	Pg	Aa	Fn	Cr	胎盘氧代谢β-actin/eNOS^c	NO(μmol/L)^d	胎盘系数	胎盘重量(g)^e
PE组		?	?	?	?	?	?	?	?	?	?
?	PE-A亚组	16	7.7±3.1	11(68.75)	9(56.00)	7(43.75)	4(25.00)	16.1±2.6	35.5±16.7	0.2±0.1	467.9±104.4
?	PE-B亚组	9	7.1±2.2	3(33.33)	3(33.33)	2(22.22)	1(11.11)	12.7±3.5	49.7±15.0	0.2±0.0	441.0± 97.3
对照组		?	?	?	?	?	?	?	?	?	?
?	对照-A亚组	10	5.8±2.3	4(40.00)	3(30.00)	2(20.00)	0(0.00)	10.8±2.6	64.3±23.6	0.2±0.0	593.4±119.9
?	对照-B亚组	17	5.9±1.9	4(23.50)	1 (5.88)	2(11.76)	0(0.00)	7.7±2.1	77.6±16.8	0.2±0.0	603.5± 83.7
t值/χ²值		?	2.0	10.1	10.98	4.695	8.647	28.3	17.2	2.7	8.8
P值		?	0.126	0.018	0.012	0.196	0.034	0.000	0.000	0.057	0.000

表4 牙周菌在胎盘组织的分布及胎盘组织的NO含量情况

Table 4 The distribution of periodontal bacteria and the metabolism of NO in placenta

组别		例数	血清CRP^a	胎盘组织中检出牙周细菌^b				胎盘氧代谢β-actin/eNOS^c	NO(μmol/L)^d	胎盘系数	胎盘重量(g)^e
组别		例数	血清CRP^a	Pg	Aa	Fn	Cr	胎盘氧代谢β-actin/eNOS^c	NO(μmol/L)^d	胎盘系数	胎盘重量(g)^e
PE组		?	?	?	?	?	?	?	?	?	?
?	PE-A亚组	16	7.7±3.1	11(68.75)	9(56.00)	7(43.75)	4(25.00)	16.1±2.6	35.5±16.7	0.2±0.1	467.9±104.4
?	PE-B亚组	9	7.1±2.2	3(33.33)	3(33.33)	2(22.22)	1(11.11)	12.7±3.5	49.7±15.0	0.2±0.0	441.0± 97.3
对照组		?	?	?	?	?	?	?	?	?	?
?	对照-A亚组	10	5.8±2.3	4(40.00)	3(30.00)	2(20.00)	0(0.00)	10.8±2.6	64.3±23.6	0.2±0.0	593.4±119.9
?	对照-B亚组	17	5.9±1.9	4(23.50)	1 (5.88)	2(11.76)	0(0.00)	7.7±2.1	77.6±16.8	0.2±0.0	603.5± 83.7
t值/χ²值		?	2.0	10.1	10.98	4.695	8.647	28.3	17.2	2.7	8.8
P值		?	0.126	0.018	0.012	0.196	0.034	0.000	0.000	0.057	0.000

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