The Effects Of GENGNIANGANLUYIN Pulsing Low-dose Estrogen On Endocrine And Target Organs Of Castrated Rats

Peri-menopausal syndrome (climacteric syndrome) occurs aroundwomen menopause including a serial of somatic and psycho-emotionaldisturbances symptom induced by sex hormone decreasing. After 40 yearsold, female will enter peri-menopausal period in which ovary function andfemale hormone secretion begin decline, accompanied by various degreemenstruation disturbance and hypo-estrogenism symptom, which bring hugedisadvantage to women's living and working. If peri-menopausal syndromecannot prevent and treated in time, many old-age diseases, such asosteoporosis, cardiovascular complications, cerebral vessels complicationsand presenile dementia will be following.In past 50 years, hormone replacement therapy (HRT) has been used toalleviate a serial of estrogen deficiency problems. However, long-termapplication can evoke breast cancer, carcinoma of endometrium, abnormalcolpo bleeding,obesity and other ill effects, although HRT can alleviateperi-menopausal period symptom. Furthermore, there are many unsolvedproblems about HRT, such as the interface of various kinds sex hormone,the action, dosage, proportion, medication duration and medicationindividualization of sex hormone, which limited the spread of HRT.Therefore, the urgent thing at present is that maintains amelioration actionof HRT and decreases ill effects both at the same time.Traditional Chinese medical science has cognized peri-menopausalsyndrome with a history of thousands years. Traditional Chinese medicine'snature is milder and it's ill effects are fewer. Methods and techniques ofmodern medicine have been used to explore the mechanics of the traditionalChinese medicine curing peri-menopausal syndrome in these decades.Several traditional Chinese medicine prescription can alleviate menopausalsymptom, Adjust endocrine system, enhance immunological function.However, the traditional Chinese medicine cannot act on estrogen receptor(ER) that distribute the whole body and cannot replace the multi-protectionof estrogen, so the result of using the traditional Chinese medicine alone onovary dysfunction and hypo-estrogenism symptom is not very satisfactory. In this study, the domestic traditional Chinese medicineprescription-GENGNIANGANLUYIN pulsing low-dose estrogen (G+E)was adopted in rat peri-menopausal syndrome model. The influence of(G+E) on rats'endocrine and target organ was investigated and comparedwith that of using GENGNIANGAN LUYIN(G) and estrogen (E)respectively. The aim of the study was to find a cure, which can maintainthe good effect of HRT and minimize the ill effects of HRT at the sametime. Methods: Explore the best level of low-dose estrogen before experiments.Choose 0.15mg/kg as selected low-dose estrogen (being equal to 1/4 ofcommon dosage using on human being), which cannot increase serumestrogen level marked and has no significant influence on endometriumstructure. 200～220g Wistar female rats were castrated and divided into 4 groupsand pseudo-operation group as control. Pseudo-operation group and modelgroup were given distilled water 10ml/kg/d; G group were givenGENGNIANGANLUYIN 10ml/kg/d; E group were given estrogen0.6mg/Kg/semi-month; G+E group were given GENGNIANGANLUYIN10ml/kg/d and 0.15mg/Kg/semi-month. Three month later, the rats werescarified. Serum CHO and TG were detected by biochemistry auto-Analysis; serum E2, FSH, LH were investigated by radio-immunity; vaginashedding cells score, endometrium morphology were evaluated by HE andelectron microscope; expression of estrogen receptor in uterus and thoracicaorta were analyzed by RT-PCR and immunohistochemistry. The results: 1. Serum hormone level Compared with pseudo-operation group, model group had a decreasedE2,increased FSH and LH in serum(P<0.01). all of Three kind of treatmentcan increase E2,decrease FSH and LH in serum(P<0.01). There were nosignificant differential between E group and G+E group, but both groupshad more obvious effects than G group(P<0.01). 2. Serum CHO and TG level Compared with pseudo-operation group, serum CHO and TG levelincreased obviously in model group. Compared with model group, threekind of treatments decrease serum CHO and TG level (P<0.01). Therewere no significant differential between E group and G+E group, but bothgroups had more obvious effects than G group(P<0.01). 3. Vagina shedding cells score After 3-month treatment, vagina-shedding cells from 5 groups weresmeared and scored. Compared with pseudo-operation group, model grouphad a decreased score(P<0.01). Compared with model group, threetreatments groups had a increased score(P<0.01). There were no significantdifferential between E group and G+E group, but both groups had moreobvious effects than G group(P<0.01). 4. Femur bone mineral density (BMD)After 3-month treatment, bone mineral density meter measured thebone mineral density of rats'left femurs. Compared with pseudo-operationgroup, model group had a decreased BMD(P<0.01). Compared with modelgroup, three treatments groups had a increased BMD(P<0.01). There wereno significant differential between E group and G+E group(P>0.05), butboth groups had more obvious effects than G group(P<0.01). 5. Endometrium morphology ①HE After 3-month treatment, endometrium atrophied in rats of modelgroup and G group. Endometrium had an apparent hyperplasia trace. Therewere not obvious atrophy and hyperplasia trace in G+E group. ②electron microscope Lissome, lipofuscin, marrow sample Corpuscle appeared in modelgroup endometrium glandular epithelium, which mark the trace of senility.There were not obvious abnormal in pseudo-operation group. 6. Estrogen receptor expression in uterus Estrogen receptor (ER) protein expression in uterus had been detectedby immunohistochemistry and been analyzed on gray degree. ERexpressed mainly in endometrium and gland and little positive stain alsoexisted in muscle cell and interstitial. Compared with pseudo-operationgroup, ER expression decreased in model group(P<0.01). Compared withmodel group, three treatments groups had a increased ER expression(P<0.01). There were no significant differential between E group and G+Egroup(P>0.05), but both groups had more obvious effects than G group(P<0.01). 7. ERαmRNA in thoracic aorta Estrogen receptor (ER) αmRNA expression in thoracic aorta had beeninvestigated by RT -PCR and been Analyzed on gray degree. Comparedwith pseudo-operation group, ER expression decreased in model group(P<0.01). Compared with model group, three treatments groups had aincreased ER expression(P<0.01), but both E and G+E groups had moreobvious effects than G group(P<0.01). The conclusion: 1. GENGNIANGANLUYIN pulsing low-dose estrogen can alleviateblood serum hormone change, decrease blood lipo, maintain estrogenreceptor content of uterus and thoracic aorta, and prevent osteoporosis. Theeffects are similar with that of estrogen and better than that ofGENGNIANGANLUYIN. 2. GENGNIANGANLUYIN pulsing low-dose estrogen doesn't makeendometrium hyperplasia obviously. GENGNIANGANLUYIN pulsing low-dose estrogen can be a newcure using on peri-menopausal syndrome.