Confirming the Diagnosis of Hyperthyroidism: Thyrotropin-Releasing Hormone (TRH) Stimulation Test

In my last blog post, I discussed use of dynamic test, the T3 suppression test. Suppression testing can be helpful in confirming the diagnosis when we suspect that a cat has mild hyperthyroidism but serum concentrations of total and free T4 are either normal or borderline.

An alternative dynamic test that can be used to help diagnose cats with mild or occult hyperthyroidism is the thyrotropin-releasing hormone (TRH) stimulation test (1-3). Compared with the T3 suppression test, this test takes the opposite approach — now we are stimulating, rather than suppressing, the secretion of TSH from the pituitary, which in turn, would lead to increased T4 secretion.

Thyrotropin-Releasing Hormone (TSH) Stimulation Test

TRH is the hormone released by the hypothalamus that subsequently stimulates the release of thyrotropin (TSH) from the pituitary gland (4). When administered intravenously to normal cats, TRH causes an prompt increase in TSH secretion and serum T4 concentrations (see Figure 1, left panel).

Figure 1: TRH tests in normal cats (left) & cats with hyperthyroidism (right)

In contrast to the situation in normal cats, administration of large doses of TRH to hyperthyroid cats has little or no effect on TSH or T4 secretion. The reason for this that pituitary TSH secretion has already been completely and chronically suppressed by the higher-than-normal thyroid hormone secretion characteristic of hyperthyroidism (see Figure 1 above, right panel).

Administration of only a single dose of TRH is not enough to stimulate chronically suppressed TSH-secreting cells.

Testing Protocol for the TRH Stimulation Test

To perform the TRH stimulation test in cats, the following protocol is recommended:

1. The veterinarian draws a blood sample for determination of baseline serum concentrations of total T4 (and T3).
2. TRH (Protirelin) is administered to the cat at the dosage of 100 μg/kg, IV.
3. Four hours later, the veterinarian again draws a blood sample for serum T4 (and T3) determinations.

One may ask: but what about measuring TSH? Indeed, that would be a more direct measure and would shorten the testing time from 4 hours to less than an hour (4). However, because use of TSH measurements in cats with thyroid disease have only recently been advocated, studies looking the serum TSH response to TRH have yet to be published in cats.

Interpretation of Results of TRH Stimulation Testing

When the TRH stimulation test is performed in normal cats and sick cats without hyperthyroidism (1), there is a consistent rise in serum T4 concentrations (approximately 2-fold rise). In contrast, when the test is performed in cats with hyperthyroidism, even in cats with only slightly high or high-normal resting serum T4 concentrations, there is little, if any, rise in serum T4 values after the administration of TRH (see Figure 2, below).

Figure 2 — Box plots of the serum T4 concentrations before (A) and after (B) TRH stimulation in 31 clinically normal cats, 35 cats with hyperthyroidism, and 15 cats with nonthyroidal disease. The "box" represents the interquartile range from the 25th to 75th percentile (represents the middle one-half of the data). The horizontal bar through the box is the median. The "whiskers" represent the main body of data, which in most cases is equal to the range. Outlying data points are represented by open circles. To convert serum T4 concentrations from nmol/L to µg/dl, divide the given values by 12.87.

Regarding interpretation of the TRH stimulation test results, we found that the relative rise (per cent increase) in serum T4 concentration after administration of TRH was the best (most sensitive) criteria for predicting whether or not cats are hyperthyroid (1). A percent rise in serum T4 of less than 50 per cent is consistent with mild hyperthyroidism, whereas a value of greater than 60 per cent is seen in normal cats; values between 50 and 60 per cent are equivocal or borderline responses.

The serum T3 response to TRH is less helpful in separating normal from hyperthyroid cats, because many normal cats have only a small and inconsistent rise in serum T3 concentrations after TRH administration. Therefore, I do not recommend determining the serum T3 response as part of the TRH stimulation test.

In another study of 36 cats with severe critical illness (5), half of the severely sick cats showed an inadequate T4 response to TRH stimulation test (i.e., a percent rise in serum T4 of less than 50 per cent). Unfortunately, this could create a situation where mild hyperthyroidism is falsely diagnosed in a cat with critical illness.  Although the reason for this is unclear, critical nonthyroidal illness may render pituitary TSH secretion unresponsive to the stimulatory effects of TRH administration.

The bottom line: This test is not meant to differentiate normal and hyperthyroid cats from cats with severe critical illness. I do not recommend doing this test in severely ill cats, inasmuch as the test results may be invalid.

Which is Better: TRH Stimulation Test vs. the T3 Suppression Test?

Advantages of the TRH stimulation test over the T3 suppression test include the shorter time needed to perform the test (4 hours vs. 3 days), and the fact that the TRH stimulation test is not dependent upon the owner's ability to administer oral medication.

The major disadvantage of the TRH stimulation test in cats is that side effects almost invariably occur immediately after administration of the TRH (1, 6-8). These adverse effect include the following:

• Excessive salivation (see Figure 3)
• Vomiting
• Induced defecation
• Rapid breathing (tachypnea); open mouth breathing

Fortunately, all of the adverse side effects are transient and completely resolve by the end of the 4-hour test period. Because of these adverse effects, however, I generally do a T3 suppression test over the TRH stimulation test for dynamic testing of the pituitary-thyroid axis.

Figure 3: Cat showing increased salivation after TRH administration

Our studies show a close relationship between the presence (or absence) of suppressed serum T4 concentrations in response to T3 suppression and stimulated T4 values in response to TRH stimulation. Therefore, although the two tests evaluate the pituitary-thyroid axis in different ways, our findings indicate that the two screening tests provide similar information and can probably be used interchangeably for diagnosing mild hyperthyroidism in cats.

Future Investigations: Serum TSH Response to TRH Stimulation in Cats

In human beings, the TRH stimulation test is generally done by directly evaluating the change in serum TSH concentrations, not indirectly by measuring serum T4  values.  When this is done, the TRH stimulation test is one of the most sensitive tests in detecting early or mild hyperthyroidism in human patients. Again, because TSH production is completely suppressed in hyperthyroid patients, they will not show a rise in serum TSH when TRH is administered, whereas normal individuals will respond with a brisk rise in circulating TSH within 30 minutes of TRH injection. The new, ultra-sensitive human TSH assays have largely eliminated the need for TRH stimulation testing, but it still remains a useful and time-proven test.

Similar serum TSH responses to TRH have been reported in normal dogs — baseline serum TSH concentrations increase 2- to 3-fold when measured 30 minutes after administration of TRH at doses ranging from 1-10 μg/kg. There are two major advantages of evaluating the TSH response rather than the T4 response to TRH:

1. The test period is shorten from 4 hours to only 15-30 minutes.
2. A much lower dose of TRH is needed, greatly reducing or eliminating the adverse signs commonly seen with the higher doses needed to elicit a serum T4 response.

Measuring the serum TSH response to TRH has not been reported in cats with hyperthyroidism. Obviously, this is an area that needs to be investigated, now that we know that the canine TSH assay can be used to measure circulating TSH in cats.

Because the canine TSH assay is not designed to measure low values, basal TSH measurements are not very useful as a diagnostic test in feline hyperthyroidism (see my previous blog on TSH in cats). However, measuring the response to TRH may turn out to be a extremely useful and sensitive test for this disease.

References:

1. Peterson ME, Broussard JD, Gamble DA: Use of the thyrotropin releasing hormone stimulation test to diagnose mild hyperthyroidism in cats. Journal of Veterinary Internal Medicine 1994;8:279-286.
2. Peterson ME. Diagnostic tests for hyperthyroidism in cats. Clinical Techniques in Small Animal Practice 2006;21:2-9.
3. Peterson ME: Diagnostic testing for feline hyper- and hypothyroidism. Proceedings of the 2011 American College of Veterinary Internal Medicine (ACVIM) Forum, pp. 95-97, 2011.
4. Utiger RD: Tests of thyroregulatory mechanisms. In Ingbar SH and Braverman LE (eds): The Thyroid: A Fundamental and Clinical Text, pp 511-523. Philadelphia, JB Lippincott, 1986.
5. Tomsa K, Glaus TM, Kacl GM, et al. Thyrotropin-releasing hormone stimulation test to assess thyroid function in severely sick cats. Journal of Veterinary Internal Medicine 2001;15:89-93.
6. Holtman JR, Buller AL, Hamosh P, et al: Central respiratory stimulation produced by thyrotropin-releasing hormone in the cat. Peptides 1986;7:207-212.
7. Beleslin DB, Jovanovic-Micic D, Tomic-Beleslin N: Nature of salivation produced by thyrotropin-releasing hormone (TRH). Brain Research Bulletin 1987;18:463-465.
8. Beleslin DB, Jovanovic-Micic D, Samardzic R, et al: Studies of thyrotropin-releasing hormone (TRH)-induced defecation in cats. Pharmacology Biochemistry & Behavior 1987;26:639-641.
9. Yagi K, Ohashi E, Tanabe S, Uzuka Y, Sarashina T. Serum thyrotropin response to TRH administration in six healthy beagle dogs. Veterinary Record 2000;146:706-707.